Assessment of stress levels and reproductive condition in giant pandas: insights from hair, faecal and saliva samples.

Concerted conservation efforts have brought the giant panda (Ailuropoda melanoleuca) back from the brink of extinction, but pandas continue to face anthropogenic threats in the wild and breeding success in captivity remains low. Because stress can have detrimental impacts on reproduction, monitoring stress- and sex-steroid levels would help assess the effectiveness of conservation mitigation measures in panda populations as well as monitor the welfare and reproductive health of captive animals. In this proof-of-concept study, we used faecal sex steroid and cortisol concentrations (n = 867 samples collected from five males and five females at Beijing Zoo every 4 days over the course of 12 months) as a reference to investigate if testosterone, estradiol, progesterone and cortisol can be meaningfully measured in panda hair (n = 10) using radio-immuno-assays. Additionally, we calculated the ratio of testosterone to cortisol (T:C ratio) for each male, which can provide a biomarker of stress and physical performance. Our findings revealed distinct monthly variations in faecal sex-steroid and cortisol concentrations, reflecting reproductive seasonality and visitor-related stress among individual pandas. Notably, the oldest male had a significantly lower T:C ratio than other males. Our results confirm that the level of sex steroids and cortisol can be assayed by panda hair, and the hair cortisol concentrations correlate significantly with that in faeces with one month lag behind (r = 0.68, P = 0.03). However, the concentrations of hormones detected in saliva are lower than those in faeces by two orders of magnitude, making it difficult to ensure accuracy. By assessing the applicability of hair, faecal and salivary sampling, we can infer their utility in monitoring the reproductive status and acute and chronic stress levels of giant pandas, thereby providing a means to gauge the success of ongoing habitat restoration efforts and to discuss the feasibility of sample collection from wild populations.

Natural mating ability is associated with gut microbiota composition and function in captive male giant pandas.

The issue of poor sexual performance of some male giant pandas seriously impairs the growth and the genetic diversity of the captive population, yet there is still no clear understanding of the cause of the loss of this ability and its underlying mechanism. In this study, we analyzed the gut microbiota and its function in 72 fecal samples obtained from 20 captive male giant pandas, with an equal allocation between individuals capable and incapable of natural mating. Additionally, we investigated fecal hormone levels and behavioral differences between the two groups. A correlation analysis was then conducted among these factors to explore the influencing factors of their natural mating ability. The results showed significant differences in the composition of gut microbiota between the two groups of male pandas. The capable group had significantly higher abundance of Clostridium sensu stricto 1 (p adjusted = .0021, GLMM), which was positively correlated with fatty acid degradation and two-component system functions (Spearman, p adjusted < .05). Additionally, the capable group showed higher gene abundance in gut microbiota function including purine and pyrimidine metabolism and galactose metabolism, as well as pathways related to biological processes such as ribosome and homologous recombination (DEseq2, p adjusted < .05). We found no significant differences in fecal cortisol and testosterone levels between the two groups, and no difference was found in their behavior either. Our study provides a theoretical and practical basis for further studying the behavioral degradation mechanisms of giant pandas and other endangered mammal species.

Giant panda (Ailuropoda melanoleuca) neonates use broadband calls to communicate with their mothers.

Infant call structure should have evolved to elicit maximum maternal attention and investment. Neonates of giant pandas produce three types of vocalizations reported to be vitally important in the context of mother-infant communications. However, how cubs, 0-15 days old, communicate with their mothers to elicit maternal care remains unknown. We analyzed 12 different call parameters of 3475 squawks, 1355 squalls, and 491 croaks from 11 captive giant panda (Ailuropoda melanoleuca) neonates from age 0 to 15 days. In playback experiments, we also tested whether mothers could detect ultrasound. Our results show that neonates use broadband calls with ultrasonic frequencies up to 65 kHz to convey information about their physiological needs and to attract maternal care. In playback experiments, we tested if mothers reacted differently to broadband calls (BBC) than to artificially altered calls that included only frequencies <20 kHz (AUDC) or calls that included only frequencies >20 kHz (USC). Playback confirmed that, although adult females responded significantly less often to USC, BBC than to or AUDC, they could detect USC, BBC and generally made appropriate behavioral responses, indicating a potential benefit for neonates to utilize ultrasonic and broadband frequencies. Our findings provide a new insight into mother-infant communication in giant pandas and will be helpful for reducing the mortality of cubs, younger than 1 month old, in captivity.

Fecal Bacterial Community of Allopatric Przewalski's Gazelles and Their Sympatric Relatives.

Mammal gastrointestinal tracts harbor diverse bacterial communities that play important roles in digestion, development, behavior, and immune function. Although, there is an increasing understanding of the factors that affect microbial community composition in laboratory populations, the impact of environment and host community composition on microbiomes in wild populations is less understood. Given that the composition of bacterial communities can be shaped by ecological factors, particularly exposure to the microbiome of other individuals, inter-specific interactions should impact on microbiome community composition. Here, we evaluated inter-population and inter-specific similarity in the fecal microbiota of Przewalski's gazelle (Procapra przewalskii), an endangered endemic ruminant around Qinghai Lake in China. We compared the fecal bacterial communities of three Przewalski's gazelle populations, with those of two sympatric ruminants, Tibetan gazelle (Procapra picticaudata) and Tibetan sheep (Ovis aries). The fecal bacterial community richness (Chao1, ACE) did not vary across the three Przewalski's gazelle populations, nor did the composition vary between species. In contrast, the managed Przewalski's gazelle population had higher bacterial diversity (Shannon and Simpson) and was more similar to its sympatric Tibetan sheep in beta diversity than the wild Przewalski's gazelle populations. These results suggest that ecological factors like host community composition or diet affect Przewalski's gazelle's gastrointestinal bacterial community. The role of bacterial community composition in maintaining gastrointestinal health should be assessed to improve conservation management of endangered Przewalski's gazelle. More broadly, captive breeding and reintroduction efforts may be impeded, where captive management results in dysbiosis and introduction of pathogenic bacteria. In free ranging populations, where wildlife and livestock co-occur, infection by domestic pathogens and diseases may be an underappreciated threat to wild animals.

Effect of group size and reproductive status on faecal glucocorticoid concentration and vigilance in a free-ranging population of Przewalski's gazelle.

Elevated glucocorticoid (GC) concentration and increased vigilance are two common responses to predation risk in mammals. Chronic high-level GC concentration and vigilance occur at the expense of other life maintenance and reproduction activities, reflecting a trade-off between individual survival and future fecundity. Przewalski's gazelle (Procapra przewalskii) is a group-living ungulate endemic to the high-altitude Qinghai Lake region of China. Group-size effect on gazelle vigilance has been examined, yet little is known about how their GC concentration is affected by group size or reproductive status. In this study, we examined the effect of group size and reproductive status on faecal glucocorticoid metabolite (FGM) concentrations and individual vigilance during different stages of the reproduction cycle (i.e. non-breeding, lambing and rutting) in free-ranging adult female Przewalski's gazelles. Group size did not influence FGMs significantly, but mean vigilance duration increased with group size. The gazelles' FGMs and vigilance peaked in lambing season. FGMs showed no difference between rutting season and non-reproductive season, but vigilance was lowest in the rutting season. FGMs correlated with vigilance frequency and vigilance duration. Antipredator responses of female Przewalski's gazelles appear to change with reproductive status but not with group size in free-ranging females. Management measures should be taken in the lambing season to minimize stress on mother gazelles.

Prospective Study on the Excretion of Mucous Stools and its Association with Age, Gender, and Feces Output in Captive Giant Pandas.

The giant panda (Ailuropoda melanoleuca) has evolved a large number of mucous glands in the intestinal lining to adapt to the digestion of high-fiber foods. However, in captive pandas, excessive mucus might form a mass and then be eliminated, which is often accompanied by discomfort and decreased activity. This event is called 'mucous excretion'. The causes of mucus excretions in captive pandas, however, remain unknown. The aims of this study were to document the occurrence of mucus excretion and to investigate its possible associations with pandas' age, gender, and feces output. Eighteen giant pandas were studied at the Beijing Zoo from April 2003 to June 2017, and a total of 900 occurrences of mucous excretion and 32,856 daily defecation outputs in weight were recorded. The likelihood of mucous excretion occurrence decreased by 11.34% for each 1 kg of fecal output (Z = -4.12, p < 0.0001), while it increased by 5.89% per year of age (Z = 4.02, p < 0.0001). However, individual differences in gender had no significant effect on the mucous occurrence (Z = -0.75, p = 0.4508). A monthly change in mucus occurrence was also found. The mean frequency of mucus occurrence was significantly higher in October. In August, time (month) change showed the biggest negative influence on feces output but the biggest positive influence on mucus excretion (seasonal factors were -2.261 and 0.0126, respectively). Our results documented the occurrence of mucous excretions and confirmed their possible associations with the pandas' age and fecal output based on a 15-year prospective study. This study not only adds to our knowledge of panda physiology but also suggests the need for further studies examining the causes of the excretion of mucous stools in captive pandas. Reducing the incidence of mucous excretion would promote ex situ conservation and enhance panda welfare.

Seasonal and reproductive variation in chemical constituents of scent signals in wild giant pandas.

Seasonally reproducing animals show many behavioral and physiological changes during the mating period, including increased signaling for intrasexual competition and mate attraction. We collected 102 anogenital gland secretions (AGS) from marking trees in Foping Nature Reserve, and used gas chromatography mass spectrometry analyze these chemical composition. Of these marks, all but one were from males, confirmed with DNA analysis. We found that several chemical constituents, especially volatile compounds, is present only during the mating season and that the relative abundance of many compounds changed as a function of breeding season, whereas nonvolatile compounds were lower in the mating season. This seasonal variation in chemical composition of AGS most likely plays an important role in governing giant panda reproduction, including mate location, attraction, and male-male competition. The chemical properties of many of these putative chemosignals-such as volatility and longevity-are suggestive of these roles, and undoubtedly contribute to successful reproduction for this species with a characteristically sophisticated chemical communication system. We also found a number of important differences between the chemical constituents of AGS from wild pandas and those found in previous studies with captive pandas, suggesting that inappropriate chemosignal composition may contribute to poor reproductive success in captive breeding programs.

Individuality in coo calls of adult male golden snub-nosed monkeys (Rhinopithecus roxellana) living in a multilevel society.

Vocal individuality is a prerequisite for individual recognition, especially when visual and chemical cues are not available or effective. Vocalizations encoding information of individual identity have been reported in many social animals and should be particularly adaptive for species living in large and complexly organized societies. Here, we examined the individuality in coo calls of adult male golden snub-nosed monkeys (Rhinopithecus roxellana) living in a large and multilevel society. Coo calls are one of the most frequently occurring call types in R. roxellana and likely serve as the signals for contact maintenance or advertisement in various contexts including group movement, foraging, and resting. From April to October 2016, April to July 2017, and September to October 2017, we recorded a total of 721 coo calls from six adult males in a provisioned, free-ranging group and one adult male in captivity in Shennongjia National Park, China. We selected 162 high-quality recordings to extract 14 acoustic parameters based on the source-filter theory. Results showed that each of all parameters significantly differed among individuals, while pairwise comparisons failed to detect any parameter that was different between all pairs. Furthermore, a discriminant function analysis indicated that the correct assignment rate was 80.2% (cross-validation: 67.3%), greater than expected by chance (14.3%). In conclusion, we found evidence that coo calls of adult male R. roxellana allowed the reliable accuracy of individual discrimination complementarily enhanced by multiple acoustic parameters. The results of our study point to the selective pressures acting on individual discrimination via vocal signals in a highly gregarious forest-living primate.

Vocal repertoire of free-ranging adult golden snub-nosed monkeys (Rhinopithecus roxellana).

Vocal signaling represents a primary mode of communication for most nonhuman primates. A quantitative description of the vocal repertoire is a critical step in in-depth studies of the vocal communication of particular species, and provides the foundation for comparative studies to investigate the selective pressures in the evolution of vocal communication systems. The present study was the first attempt to establish the vocal repertoire of free-ranging adult golden snub-nosed monkeys (Rhinopithecus roxellana) based on quantitative methods. During 8 months in Shennongjia National Park, China, we digitally recorded the vocalizations of adult individuals from a provisioned, free-ranging group of R. roxellana across a variety of social-ecological contexts. We identified 18 call types, which were easily distinguishable by ear, visual inspection of spectrograms, and quantitative analysis of acoustic parameters measured from recording samples. We found a great sexual asymmetry in the vocal repertoire size (females produced many more call types than males), likely due to the sex differences in body size and social role. We found a variety of call types that occurred during various forms of agonistic and affiliative interactions at close range. We made inference about the functions of particular call types based on the contexts in which they were produced. Studies on the vocal communication in R. roxellana are particularly valuable since they provide a case about how nonhuman primates, inhabiting forest habitats and forming complex social systems, use their vocalizations to interact with their social and ecological environments.

Diversity of Thermogenic Capacity Predicts Divergent Obesity Susceptibility in a Wild Rodent.

OBJECTIVE: The objective of the present study was to examine whether wild rodents exhibit diverse obesity susceptibility and what factors predispose subjects to this divergence in response to a high-fat diet (HFD). METHODS: Sixty male and female Brandt's voles (Lasiopodomys brandtii) were fed an HFD for 8 weeks, and the upper (obesity prone [OP]) and lower (obesity resistant [OR]) one-third for mass gain were selected. Energy budgets and pathologic changes were measured. Another 30 males were fed a low-fat control diet (LFD) for 10 weeks and then fed an HFD for 12 weeks. The energetic parameters of the rodents on an LFD were analyzed for the correlation with body mass of the rodents on an HFD. RESULTS: OP voles had higher energy intakes, higher levels of noradrenaline-induced nonshivering thermogenesis, and a greater impairment of insulin tolerance than OR voles. Unlike laboratory rodents, there were no differences in physical activity or resting metabolic rate between these groups of voles. The thermogenic capacity during LFD feeding was the strongest predictor for mass gain during HFD feeding. CONCLUSIONS: This study suggests that a wild rodent species of Brandt's voles exhibits diverse obesity susceptibility in reaction to an HFD, providing a natural model to give insight into the mechanisms for divergent obesity susceptibility. This study also indicates that maximum thermogenic capacity has a predictive power for the development of obesity when an HFD was available.

Reverse chemical ecology: Olfactory proteins from the giant panda and their interactions with putative pheromones and bamboo volatiles.

The giant panda Ailuropoda melanoleuca belongs to the family of Ursidae; however, it is not carnivorous, feeding almost exclusively on bamboo. Being equipped with a typical carnivorous digestive apparatus, the giant panda cannot get enough energy for an active life and spends most of its time digesting food or sleeping. Feeding and mating are both regulated by odors and pheromones; therefore, a better knowledge of olfaction at the molecular level can help in designing strategies for the conservation of this species. In this context, we have identified the odorant-binding protein (OBP) repertoire of the giant panda and mapped the protein expression in nasal mucus and saliva through proteomics. Four OBPs have been identified in nasal mucus, while the other two were not detected in the samples examined. In particular, AimelOBP3 is similar to a subset of OBPs reported as pheromone carriers in the urine of rodents, saliva of the boar, and seminal fluid of the rabbit. We expressed this protein, mapped its binding specificity, and determined its crystal structure. Structural data guided the design and preparation of three protein mutants bearing single-amino acid replacements in the ligand-binding pocket, for which the corresponding binding affinity spectra were measured. We also expressed AimelOBP5, which is markedly different from AimelOBP3 and complementary in its binding spectrum. By comparing our binding data with the structures of bamboo volatiles and those of typical mammalian pheromones, we formulate hypotheses on which may be the most relevant semiochemicals for the giant panda.

Preliminary study on hand preference in captive northern white-cheeked gibbons (Nomascus leucogenys).

The postural origin hypothesis and the task complexity hypothesis propose that hand preference in non-human primates evolved in association with body posture and task complexity, respectively. The results of previous studies testing these two hypotheses, however, vary greatly with the different primate species and methods used. To investigate the effect of body posture and task complexity on hand preference, we recorded bouts of hand usage in nine captive northern white-cheeked gibbons (Nomascus leucogenys) housed at Beijing Zoo as they reached for food items in a ground-reaching task, a box task, and a tube task. The results showed that four to seven of the nine gibbons displayed a hand preference at the individual level in different tasks, and that hand preference in individuals was task-specific; there was no group-level hand preference in any task. The box task seemed to elicit a greater strength of hand preference than the ground-reaching task at the individual level. Although the small sample size rules out drawing any strong conclusions concerning hand preference at the group level, our results suggest that the suspensory reaching posture might increase the expression of hand preference at the individual level. This study provides preliminary information on hand preference in captive northern white-cheeked gibbons, and will be helpful for future studies addressing the origin and evolution of hand preference in small apes.

Phylogenetic studies of Prunella strophiata (Passeridae: Prunella) based on complete mitochondrial DNA sequences.

Rufous-breasted Accentor (Prunella strophiata) is a small-sized bunting with an extremely geographical range in the world. Here, the complete mitochondrial genome of P. strophiata (16,830 bp in length) has been analyzed for building the database. Similar to the typical mtDNA of vertebrates, it contained 37 genes (13 protein-coding genes, two rRNA genes and 22 tRNA genes) and a non-coding region (D-loop). Overall base composition of the complete mitochondrial DNA is A (30.3%), G (14.7%), C (31.0%) and T (24.0%), the percentage of A and T (54.3%) is higher than G and C (45.7%). All the genes in P. strophiata were distributed on the H-strand, except for the ND6 subunit gene and 9 tRNA genes which were encoded on the L-strand. The phylogenetic relationships of 12 Passeriformes species were reconstructed based on the complete mtDNA sequences using the Bayesian inference method.

Hormones orchestrated pre- and post-copulatory sexual traits in male Mongolian gerbils.

Parker's sperm competition model predicts a negative relationship between pre-copulatory (social status) and post-copulatory (sperm quality and quantity) sexually selected traits, however, empirical studies have revealed considerable inconsistency in this relationship. We hypothesized that there was a trade-off between pre- and post-copulatory sexually selected traits, and hormones (corticosterone, CORT; testosterone, T) orchestrate this relationship. In this study, we measured energetic parameters in the dominant-subordinate Mongolian gerbils (Meriones unguiculatus), whose relationship was established under chronic social encounters in a neutral arena, and then tested the relationship between their social status and sperm quality and quantity. Our results showed that dominant males initiated attack sooner and displayed more aggression, self-grooming and locomotion behaviors in daily social encounters across seven consecutive days. Dominant gerbils also had more and better quality of sperm than that of subordinate males, yet showed no significant differences in energy intake and RMR in comparison with subordinate individuals. In addition, dominant males had higher concentrations of serum T than subordinate males, whereas the concentrations of CORT showed a reverse pattern. The frequency and duration of aggression (indicative of social status) increased with elevated T concentrations. Sperm quality in terms of number and activity were associated with higher concentrations of serum T in dominant gerbils, whereas small sperm counts and poor-quality sperm were associated with relatively higher concentrations of serum CORT in subordinate gerbils. Together, our data indicated that there was no trade-off between pre- and post-copulatory sexually selected traits but hormones orchestrated the relationship between these traits in male Mongolian gerbils.

Giant pandas failed to show mirror self-recognition.

Mirror self-recognition (MSR), i.e., the ability to recognize oneself in a mirror, is considered a potential index of self-recognition and the foundation of individual development. A wealth of literature on MSR is available for social animals, such as chimpanzees, Asian elephants and dolphins, yet little is known about MSR in solitary mammalian species. We aimed to evaluate whether the giant panda can recognize itself in the mirror, and whether this capacity varies with age. Thirty-four captive giant pandas (F:M = 18:16; juveniles, sub-adults and adults) were subjected to four mirror tests: covered mirror tests, open mirror tests, water mark control tests, and mark tests. The results showed that, though adult, sub-adult and juvenile pandas exposed to mirrors spent similar amounts of time in social mirror-directed behaviors (χ(2) = 0.719, P = 0.698), none of them used the mirror to touch the mark on their head, a self-directed behavior suggesting MSR. Individuals of all age groups initially displayed attacking, threatening, foot scraping and backwards walking behaviors when exposed to their self-images in the mirror. Our data indicate that, regardless of age, the giant pandas did not recognize their self-image in the mirror, but instead considered the image to be a conspecific. Our results add to the available information on mirror self-recognition in large mammals, provide new information on a solitary species, and will be useful for enclosure design and captive animal management.

Vomeronasal organ lesion disrupts social odor recognition, behaviors and fitness in golden hamsters.

Most studies support the viewpoint that the vomeronasal organ has a profound effect on conspecific odor recognition, scent marking and mating behavior in the golden hamster (Mesocricetus auratus). However, the role of the vomeronasal organ in social odor recognition, social interaction and fitness is not well understood. Therefore, we conducted a series of behavioral and physiological tests to examine the referred points in golden hamster. We found that male hamsters with vomeronasal organ lesion showed no preference between a predator odor (the anal gland secretion of the Siberian weasels (Mustela sibirica) and putative female pheromone components (myristic acid and palmitic acid), but were still able to discriminate between these 2 kinds of odors. In behavioral tests of anxiety, we found that vomeronasal organ removal causes female hamsters to spend much less time in center grids and to cross fewer center grids and males to make fewer crossings between light and dark boxes than sham-operated controls. This indicates that a chronic vomeronasal organ lesion induced anxious responses in females. In aggressive behavioral tests, we found that a chronic vomeronasal organ lesion decreased agonistic behavior in female hamsters but not in males. The pup growth and litter size show no differences between the 2 groups. All together, our data suggested that vomeronasal organ ablation disrupted the olfactory recognition of social chemosignals in males, and induced anxiety-like and aggressive behavior changes in females. However, a vomeronasal organ lesion did not affect the reproductive capacity and fitness of hamsters. Our studies may have important implications concerning the role of the vomeronasal organ in golden hamsters and also in rodents.

Exposure to odors of rivals enhances sexual motivation in male giant pandas.

Males will alter their mating behavior to cope with the presence of their competitors. Even exposure to odors from potential competitors can greatly increase male ejaculate expenditure in a variety of animals including insects, fishes, birds and rodents. Major efforts have been made to examine males' plastic responses to sperm competition and its fitness benefits. However, the effects of competitor absence on male's sexual motivation and behaviors remain unclear, which has been proposed to be one of the causes for the poor sexual performance of some captive mammals. This study revealed that sexual motivation can be greatly enhanced in captive male giant pandas (Ailuropoda melanoleuca) by exposure to chemosensory cues from either one or three conspecifics males. It had been shown that potential rivals' odors increased males' chemosensory investigation behavior, as well as their observing, following and sniffing behaviors towards estrous females. Behaviors changed regardless of the number of rivals (one or three). Our results demonstrate the effects of potential competition on male giant pandas' sexual motivation and behavioral coping strategy. We anticipate that our research will provide a fresh insight into the mechanisms underlying poor sexual performance in male captive mammals, and valuable information for the practical management and ex situ conservation of endangered species.

Low incidence of miscarriage induced by the scent of male littermates of original mates: male kinship reduces the bruce effect in female mice, Mus musculus.

The scent of a novel male can elicit pregnancy block in recently mated female mice (Mus musculus), a phenomenon known as the Bruce effect. Despite abundant literature on the Bruce effect in rodents, it remains unclear whether males related to a female's original mate can induce the Bruce effect in out-bred, communally living mice. We investigated this question using Kunming (KM) male mice of varying genetic relatedness. Recently mated females were subjected to three treatments: exposure to the urine of the mate, urine of the mate's male littermate, and urine of a male unrelated to the mate. It was found that the urine of male littermates of the females' mates did not elicit more pregnancy block than that of the females' mates. However, the urine of novel males caused a higher rate of female miscarriage than that of the females' mates. By using a habituation-dishabituation paradigm, we found that unmated females could discriminate the urine scents of two male littermates from those of a novel male unrelated to the littermates. To understand how females use urinary cues to discriminate between males with different genetic relationships, we used gas chromatography coupled with mass spectrometry (GC-MS) to examine the volatile composition of urine from males with varying relatedness. It was found that KM male littermates shared similar volatile compositions in their urine. Our results suggest that male kinship reduces the Bruce effect in female KM mice, and provide additional evidence for mate choice being partly mediated by the Bruce effect in KM mice.

Individual recognition and odor in rat-like hamsters: behavioral responses and chemical properties.

Individual recognition has been studied across a number of taxa and modalities; however, few attempts have been made to combine chemical and biological approaches and arrive at a more complete understanding of the use of secretions as signals. We combined behavioral habituation experiments with gas chromatography-mass spectrometry of glandular secretions from the left and right flank gland and midventral gland of the rat-like hamster, Tscheskia triton. We found that females became habituated to one scent and then could discriminate individuals via another scent source from the same individual only when familiar with the scent donor. However, this prior social interaction was not required for females to discriminate different individuals in single-stimulus habituation-dishabituation tests. Chemical analyses revealed a similarity in volatile compounds between the left and right flank gland and midventral gland scents. It appears that individually distinctive cues are integratively coded by a combination of both flank gland and midventral gland secretions, instead of a single scent, albeit animals show different preferences to the novel scent. Our results suggest that odors from the flank and midventral glands may provide information related to individuality and aid individual recognition in this species and confirm that prior interaction between individuals is a prerequisite for rat-like hamsters to form multi-odor memory of a particular conspecific.

Enhancing captive breeding in giant pandas (Ailuropoda melanoleuca): maintaining lactation when cubs are rejected, and understanding variation in milk collection and associated factors.

From 1997 to 2002, a female giant panda (Ailuropoda melanoleuca) was artificially stimulated and lactation was maintained, after her neonates were removed due to the female's inability to provide maternal care. Milk samples were collected and the amount of milk collected was quantified. The lactation curve of this animal was estimated based on the Gamma function: Y(t)=at(b)e(-ct). The amount of milk collected showed significant, positive relationships with the number of days after parturition both in 1999 and in the whole study period from 1998 to 2002. This female's lactation curves fit the type I pattern of a typical mammalian lactation curve. Daily milk collection (g) during the first 30 days after parturition, and from 31 to 60 days after parturition, showed a consistent pattern with one peak at around 8:00 hr. More milk was collected during the latter period than during the former period. The amount of milk (g) collected on mucus excretion days was significantly less than that on days after mucus excretion had ended, yet no significant difference was found between milk collected one day before mucus days and on mucus days, or between milk collected one day before and one day after mucus days. Mucus excretion from the gastrointestinal tract significantly impacted the amount of milk collected. The results from this study may aid the captive propagation and conservation of giant pandas and other endangered and rare captive mammal species.