Detection of tuberculosis in cynomolgus macaques (Macaca fascicularis) using a supplementary Monkey Interferon Gamma Releasing Assay (mIGRA).

Cynomolgus monkeys (Macaca fascicularis; MF) are commonly used as nonhuman primate models for pharmaceutical product testing. In their habitat range, monkeys have close contact with humans, allowing the possibility of bidirectional transmission of tuberculosis (TB) between the two species. Although the intradermal tuberculin skin test (TST) is used for TB detection in MF, it has limitations. Herein, we established the mIGRA, combining human QuantiFERON-TB Gold-Plus and monkey IFN-γ ELISApro systems, and used it to investigate 39 captive MF who were cage-mates or lived in cages located near a monkey who died from the naturally TB infection. During a 12-month period of study, 14 (36%), 10 (26%), and 8 (21%) monkeys showed TB-positive results using the mIGRA, the TST, and TB culture, respectively. Among the 14 mIGRA-positive monkeys, 8 (57.1%) were TST-positive and 7 (50%) were culture-positive, indicating early TB detection in the latent and active TB stages with the mIGRA. Interestingly, 3 (37.5%) of the TST-negative monkeys were culture-positive. Our study showed that the mIGRA offers many advantages, including high sensitivity and high throughput, and it requires only one on-site visit to the animals. The assay may be used as a supplementary tool for TB screening in MF.

Analysis of wild macaque stone tools used to crack oil palm nuts.

The discovery of oil palm (Elaeis guineensis) nut-cracking by wild long-tailed macaques (Macaca fascicularis) is significant for the study of non-human primate and hominin percussive behaviour. Up until now, only West African chimpanzees (Pan troglodytes verus) and modern human populations were known to use stone hammers to crack open this particular hard-shelled palm nut. The addition of non-habituated, wild macaques increases our comparative dataset of primate lithic percussive behaviour focused on this one plant species. Here, we present an initial description of hammerstones used by macaques to crack oil palm nuts, recovered from active nut-cracking locations on Yao Noi Island, Ao Phang Nga National Park, Thailand. We combine a techno-typological approach with microscopic and macroscopic use-wear analysis of percussive damage to characterize the percussive signature of macaque palm oil nut-cracking tools. These artefacts are characterized by a high degree of battering and crushing on most surfaces, which is visible at both macro and microscopic levels. The degree and extent of this damage is a consequence of a dynamic interplay between a number of factors, including anvil morphology and macaque percussive techniques. Beyond the behavioural importance of these artefacts, macaque nut-cracking represents a new target for primate archaeological investigations, and opens new opportunities for comparisons between tool using primate species and with early hominin percussive behaviour, for which nut-cracking has been frequently inferred.

An analysis of white-handed gibbon male song reveals speech-like phrases.

OBJECTIVES: Our goal was to document song phrases of the white-handed gibbon (Hylobates lar), an Asian ape that produces elaborate songs, often in well-coordinated male/female duets. We focused on the male coda, which is produced during vocal turn-taking with one's mate, and particularly its phrases containing rapid spectral and temporal variation, to investigate if modulation rates resemble those of lip-smacking in other nonhuman primates and human speech rhythm. MATERIALS AND METHODS: We produced recordings from a large population of wild gibbons. Using terminology consistent with that used to describe vocalizations in other singing species, we analyzed coda phrases, overall coda properties, coda distinctiveness across individuals, and flexibility of phrase production within song bouts. RESULTS: Our song phrase-level analysis showed that male codas differed between individuals and increase in complexity within song bouts by the addition of the only two male-specific phrases of the species' repertoire. These phrases differ from all others of the species and from vocalizations typical of the larger, nonhuman great apes, in that they contain rapid within-phrase modulation. Their modulation rates (6.82 and 7.34 Hz) are similar to that of lip-smacking in other nonhuman primates and speech in humans and, like human speech, are produced exclusively during exhalation. One phrase type (trills) contains multiple notes per exhalation, another characteristic similar to speech but not most primate vocalizations. DISCUSSION: Our data highlight the complexity and flexibility of gibbon song, and show that particular phrase features likely arose from sexual selection pressures and possess similarities to human speech rhythm.

Age related decline in female lar gibbon great call performance suggests that call features correlate with physical condition.

BACKGROUND: White-handed gibbons (Hylobates lar) are small Asian apes known for living in stable territories and producing loud, elaborate vocalizations (songs), often in well-coordinated male/female duets. The female great call, the most conspicuous phrase of the repertoire, has been hypothesized to function in intra-sexual territorial defense. We therefore predicted that characteristics of the great call would correlate with a caller's physical condition, and thus might honestly reflect resource holding potential (RHP). Because measurement of RHP is virtually impossible for wild animals, we used age as a proxy, hypothesizing that great call climaxes are difficult to produce and maintain over time, and that older adults will therefore perform lower quality great calls than young adults. To test this we analyzed the great call climaxes of 15 wild lar gibbon females at Khao Yai National Park, Thailand and 2 captive females at Leo Conservation Center, Greenwich, CT. RESULTS: Findings show that call climaxes correlate with female age, as young animals (n = 8, mean age: 12.9 years) produced climaxes with a higher frequency range (delta F0), maximum F0 frequency and duty cycle than old animals (n = 9, mean age: 29.6 years). A permuted discriminant function analysis also correctly classified calls by age group. During long song bouts the maximum F0 frequency of great call climaxes' also decreased. Additional data support the hypothesis that short high notes, associated with rapid inhalation as an individual catches its breath, reflect increased caller effort. Older females produced more high notes than younger females, but the difference only approached statistical significance, suggesting that calling effort may be similar across different ages. Finally, for the first time in this species, we measured peak intensity of calls in captive females. They were capable of producing climaxes in excess of 100 dB at close range (2.7 m). CONCLUSIONS: Age and within-bout differences in the lar gibbon great call climax suggest that call features correlate with physical condition and thus the call may have evolved as an honest signal in the context of intra-sexual territorial defense and possibly also in male mate choice via sexual selection, although further testing of these hypotheses is necessary.

Lar gibbon (Hylobates lar) great call reveals individual caller identity.

Gibbons (family Hylobatidae) produce loud, elaborate vocalizations (songs), often in well-coordinated male/female duets. The female's great call, the most conspicuous phrase of the gibbon vocal repertoire, functions primarily to mediate territorial defense. Despite the fact that lar gibbons (Hylobates lar) are the most widely distributed and well researched hylobatid species and produce a rich vocal repertoire, the individual-specificity of their great calls has not previously been quantified. In addition, spectral and temporal features of notes occurring at specific locations within the lar great call have not been described. Here we provide such a description, and test the hypothesis that great calls are statistically discriminable between a large sample of individual callers. We compared recordings of great calls from 14 wild lar females in Khao Yai National Park, Thailand. Our analyses of principal components derived from spectral and temporal measures, as well as spectrograms from the entire great call, indicate that acoustic variation is sufficient to allow identification of individual callers (83.5% discriminability based on principal components, and inter-individual call variation exceeding intra-individual variation in overall spectrogram). These vocalizations potentially allow individual recognition of animals.

Luteinizing hormone reduction by the male potency herb, Butea superba Roxb.

To determine if Butea superba Roxb., a traditional Thai male potency herb, has androgenic activity in 60-day-old male Wistar rats, we measured its effects on the pituitary-testicular axis and sex organs. Intact and orchidectomized adult male rats were subdivided into five groups (10 rats/group): distilled water, Butea superba (BS)-10, BS-50, BS-250, and testosterone propionate (TP). They received 0, 10, 50, and 250 mg·kg body weight-1·day-1 BS in distilled water by gavage and 6 mg·kg body weight-1·day-1 TP sc, respectively, during the 30-day treatment period. Blood was collected every 15 days and luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone were measured. Changes of weight and histological appearance of sex organs were determined at the end of the 30-day treatment and 15-day post-treatment periods. TP treatment reduced serum FSH and LH levels and significantly increased the weight of the seminal vesicles and epididymis, in accordance with histopathological changes, in both intact and orchidectomized rats. No changes in serum testosterone, LH, and FSH levels were observed in any of the intact rats treated with BS, but a significant increase in seminal vesicle weight was observed only in the BS-250 group. Although a significant reduction in serum LH was detected in the BS-50 and BS-250 groups of orchidectomized rats, no significant change in weight or histology of sex organs was observed. Thus, we conclude that B. superba needs endogenous testosterone to work synergistically to stimulate the accessory sex organ of intact animals and can potentially exhibit an LH reduction effect in orchidectomized animals.

Luteinizing hormone reduction by the male potency herb, Butea superba Roxb.

To determine if Butea superba Roxb., a traditional Thai male potency herb, has androgenic activity in 60-day-old male Wistar rats, we measured its effects on the pituitary-testicular axis and sex organs. Intact and orchidectomized adult male rats were subdivided into five groups (10 rats/group): distilled water, Butea superba (BS)-10, BS-50, BS-250, and testosterone propionate (TP). They received 0, 10, 50, and 250 mg·kg body weight(-1)·day(-1) BS in distilled water by gavage and 6 mg·kg body weight(-1)·day(-1) TP sc, respectively, during the 30-day treatment period. Blood was collected every 15 days and luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone were measured. Changes of weight and histological appearance of sex organs were determined at the end of the 30-day treatment and 15-day post-treatment periods. TP treatment reduced serum FSH and LH levels and significantly increased the weight of the seminal vesicles and epididymis, in accordance with histopathological changes, in both intact and orchidectomized rats. No changes in serum testosterone, LH, and FSH levels were observed in any of the intact rats treated with BS, but a significant increase in seminal vesicle weight was observed only in the BS-250 group. Although a significant reduction in serum LH was detected in the BS-50 and BS-250 groups of orchidectomized rats, no significant change in weight or histology of sex organs was observed. Thus, we conclude that B. superba needs endogenous testosterone to work synergistically to stimulate the accessory sex organ of intact animals and can potentially exhibit an LH reduction effect in orchidectomized animals.

Evidence for morphine-induced galactorrhea in male cynomolgus monkeys.

We evaluated the effect of a daily injection of morphine hydrochloride on galactorrhea in male cynomolgus monkeys. Three groups of three monkeys (nine total) were used. The treatment schedule was separated into three periods: pre-treatment, treatment, and post-treatment. Each group of monkeys was subcutaneously injected daily with 1.5, 3.0, or 6.0 mg/kg (monkey weight) of morphine for 74-130 days, respectively, during the treatment period, and with saline during the pre-treatment and post-treatment periods. We then measured the prolactin (PRL) and testosterone (T) levels from weekly blood samples that were taken 20 hours after injection. No statistically significant differences in either the PRL or T level were detected throughout the treatment period. However, monkeys treated with 3.0 and 6.0 mg/kg/day showed a decrease in T level and an increase in PRL level during the early post-treatment period. Seven of the nine monkeys produced a milk-like secretion from their mammary gland (a symptom of galactorrhea) during the treatment and post-treatment periods. For several months of post-treatment period (average 6.75 months), we monitored the time-course changes in PRL and T levels in all monkeys for 10 hours after a single injection of morphine at the same dose given during the treatment period. Morphine induced a sudden increase in the PRL level (peaked within 30 minutes) and a gradual decrease in the T level (leveled off within 6.5-10 hours), and then returned to basal levels. These results indicate that morphine does not cause a long-term effect on hormonal changes and that a morphine-induced transient rise in PRL levels accompanied by a decrease in T levels can induce spontaneous galactorrhea in male cynomolgus monkeys.

Effects of single and multiple injections of ketamine hydrochloride on serum hormone concentrations in male cynomolgus monkeys.

Using simulated short- and long-term effect studies, we evaluated the effect of ketamine anesthesia on serum cortisol, testosterone, and immunoreactive luteinizing hormone (ILH) and bioactive LH (BioLH) concentrations in adult male cynomolgus monkeys. Cortisol, testosterone, and ILH were measured by use of radioimmunoassay, and BioLH was measured by use of a radioreceptor assay method. For the acute effect, the first group (eight monkeys) was given four successive intramuscular injections of ketamine (10, 5, 5, and 5 mg/kg of body weight at 0, 30, 60, and 110 min respectively). Blood samples were taken at 0, 15, 30, 45, 60, and 120 min. For the long-term effect, the second group (10 monkeys) was given a single injection of ketamine (10 mg/kg) once a week for 4 consecutive weeks. Blood samples were taken 5 to 10 min after each injection, then were used to determine the variation in hormone concentrations among the monkeys (inter-individual variation) and within each monkey (intra-individual variation). There were no statistically significant differences in serum cortisol, testosterone, ILH, and BioLH values between the first blood sample (before the ketamine injection) and sequential blood samples in monkeys of the first group. Although intra-individual variation in the hormones (i.e., hormonal change within each monkey) was not statistically significant, inter-individual variation (among the monkeys) was significantly (0.00001 < P < 0.033) different in monkeys of the second group. These results indicate that an adequate number of animals must be used to minimize animal-to-animal variability. Our results confirm that ketamine is a suitable anesthetic agent to immobilize male cynomolgus monkeys in experimental studies (short- and long-term studies) aimed at elucidating hormonal changes.