A novel method for automatically analysing the activity of fast-moving animals: a case study of Callimico goeldii monkeys housed in a zoological garden.

Behavioural indices are recognised as important criteria for assessing animal welfare. One of the basic animal behaviours included in ethograms is their activity. The assessment of fast-moving animals, performed by humans using the visual observation method, is difficult and not very objective. Therefore, the aim of the research was to develop a method of automated analysis of animal activity, particularly useful in the observation of quick and lively individuals, and to prove its suitability for assessing the behaviour of fast-moving animals. A method of automatically assessing animal activity was developed using digital image analysis, with the Python programming language and the OpenCV library being the foundational tools. The research model was Callimico goeldii monkeys housed in a zoological garden. This method has been proved to correlate well (Rs = 0.76) with the visual method of animal behaviour analysis. The developed automatic evaluation of animal behaviour is many times faster than visual analysis, and it enables precise assessment of the daily activity of fast-moving groups of animals. The use of this system makes it possible to obtain an activity index with sub-second resolution, which allows it to be used in online mode as a detector of abnormal animal activity, e.g. early detection of illnesses or sudden events that are manifested by increased or decreased activity in relation to the standard activity pattern.

GLOBAL RETROSPECTIVE ANALYSIS OF PATHOLOGICAL FINDINGS IN ZOO-MANAGED GOELDI'S MONKEYS (CALLIMICO GOELDII), 1965-2018.

As part of the collaborative efforts and goals of managing zoo-housed Goeldi's monkeys, or callimicos (Callimico goeldii), a retrospective review of gross and histopathological postmortem examination reports submitted to the International Studbook Coordinator was carried out by veterinary representatives of the Species Survival Plan to investigate disease trends. A total of 1,887 postmortem reports (1965-2018) collected from more than 150 institutions were reviewed. Histologic findings from 862 postmortem reports and primary causes of mortality were compiled to determine the most common findings. Within the study population, 419 individuals (48.6%) were male, 383 (44.4%) female, and the remaining 60 (7%) of undetermined sex. The primary lesion at death in adults was chronic renal disease. The other prevalent lesions included cardiac disease, myelolipomas, enteritis, colitis, and hepatitis. In Great Britain and mainland Europe, Yersinia spp. infection had significantly higher prevalence than in North American callimico populations. Multiple lesions affecting more than one organ system were identified in many animals of this study population. Results also showed that for adult callimicos in zoological institutions in North America, Europe, and Great Britain, life span has been increasing over the last 50 yr.

Characterization of the phylogenetic diversity of two novel species belonging to the genus Bifidobacterium: Bifidobacterium cebidarum sp. nov. and Bifidobacterium leontopitheci sp. nov.

Two Bifidobacterium strains, i.e., 2176BT and 2177BT, were isolated from Golden-Headed Lion Tamarin (Leontopithecus chrysomelas) and Goeldi's monkey (Callimico goeldii). Isolates were shown to be Gram-positive, non-motile, non-sporulating, facultative anaerobic and d-fructose 6-phosphate phosphoketolase-positive. Phylogenetic analyses based on 16S rRNA sequences, multilocus sequences (including hsp60, rpoB, dnaJ, dnaG and clpC genes) and the core genome revealed that bifidobacterial strains 2176BT and 2177BT exhibit close phylogenetic relatedness to Bifidobacterium felsineum DSM 103139T and Bifidobacterium bifidum LMG 11041T, respectively. Further genotyping based on the genome sequence of the isolated strains combined with phenotypic analyses, clearly show that these strains are distinct from each of the type strains of the so far recognized Bifidobacterium species. Thus, Bifidobacterium cebidarum sp. nov. (2176BT=LMG 31469T=CCUG 73785T) and Bifidobacterium leontopitheci sp. nov. (2177BT=LMG 31471T=CCUG 73786T are proposed as novel Bifidobacterium species.

Angiostrongylus cantonensis central nervous system infection in captive callitrichids in Texas.

Angiostrongylus cantonensis is a tropical lungworm of rats known for central nervous system migration in aberrant primate hosts. Here, we describe A. cantonensis infection in three captive callitrichids from a Texas zoo. The affected animals included a Goeldi's monkey (Callimico goeldii), a cotton-top tamarin (Saguinus oedipus), and a pied tamarin (Saguinus bicolor) that ranged from 8 to 18 years old. Clinical signs included lethargy, ataxia, and seizures. Histologically, these animals had an eosinophilic meningoencephalitis to myelitis and some areas had abundant macrophages and parasite migration tracts. All cases had intralesional metastrongyle parasites, and nematodes were extracted from the formalin-fixed brain specimen in one case. This extracted parasite was identified as Angiostrongylus cantonensis based on morphologic features and diagnosis was confirmed with PCR. These cases represent the first report of this parasite in non-human primates in Texas, highlighting the western spread of A. cantonensis in the continental United States.

Outer retinal degeneration in two closely related Goeldi's monkeys (Callimico goeldii).

This case report comprises studies of four Goeldi's monkeys (Callimico goeldii) from the same enclosure. Globe samples from two related C goeldii (the female C goeldii and her male offspring) were available for a histopathological evaluation. Both cases presented histopathologically evident outer retinal degeneration with differences in severity. There was marked outer retinal atrophy characterized by loss of the outer and inner photoreceptor segments, and depletion of the outer retinal nuclear layer. Furthermore, we report a reduction in the thickness of the outer retinal plexiform, inner retinal nuclear layer, and inner retinal plexiform layer in these C goeldii monkeys. To the authors' knowledge, these findings have not yet been reported in wild- or captive-bred population of C goeldii.

CUTANEOUS DEMODICOSIS AND UV-INDUCED SKIN NEOPLASIA IN TWO GOELDI'S MONKEYS (CALLIMICO GOELDII).

Two nonrelated Goeldi's monkeys (Callimico goeldii) from the same enclosure developed multifocal alopecia with hyperkeratotic to ulcerative skin lesions on the lower abdomen and inner thighs. Necropsy samples of the first animal showed hyperplastic dermatitis together with in situ carcinoma and intralesional Demodex organisms. The second monkey developed similar lesions 2.5 yr later. Skin scrapings and biopsies also revealed Demodex mites within hyperplastic dermatitis. Long-term treatment with ivermectin, imidacloprid-moxidectin, and sarolaner resolved the demodicosis but skin lesions progressed to actinic keratosis and carcinoma. Both cutaneous neoplasia and demodicosis are rarely described in New World monkeys and these are the first reported cases in Goeldi's monkeys. Since the animals had access to ultraviolet (UV) light, as recommended for indoor-housed callitrichids, the skin tumors were likely UV-induced and the mites have settled particularly within impaired regions. Thus, apparent demodicosis can indicate cutaneous immunosuppression and might alert caretakers to adjust the UV regime.

Isolation of novel gut bifidobacteria using a combination of metagenomic and cultivation approaches.

Whole metagenome shotgun (WMGS) sequencing is a method that provides insights into the genomic composition and arrangement of complex microbial consortia. Here, we report how WMGS coupled with a cultivation approach allows the isolation of novel bifidobacteria from animal fecal samples. A combination of in silico analyses based on nucleotide and protein sequences facilitate the identification of genetic material belonging to putative novel species. Consequently, the prediction of metabolic properties by in silico analyses permits the identification of specific substrates that are then employed to isolate these species through a cultivation method.

COMPARISON OF TRANSCUTANEOUS ULTRASOUND AND COMPUTED TOMOGRAPHY EVALUATION OF KIDNEY SIZE AND EVALUATION OF ADRENAL GLAND SIZE USING ULTRASOUND IN A COLONY OF CALLIMICOS ( CALLIMICO GOELDII).

Both kidney and adrenal gland disease have been identified in callimicos ( Callimico goeldii). Ultrasonography (US) and computed tomography (CT) are routinely utilized in veterinary patients with suspected renal or adrenal disease to determine size, shape, and echogenicity of these organs. No previous US and CT kidney and adrenal gland measurements have been published for callimicos. In this study, 14 callimicos were anesthetized using isoflurane via facemask to evaluate kidney and adrenal gland size using US for both organs and CT for kidneys. Animals were considered clinically healthy based on history, physical examination, hematology, serum chemistry, urinalysis, and abdominal US. Ultrasound organ measurements for length (L), width (W), and height (H) in centimeters (mean/median, 95% confidence interval) in clinically healthy animals were right kidney (L = 1.90, 1.76-2.01; W = 1.05, 0.97-1.13; H = 1.59, 1.48-1.69), left kidney (L = 1.84, 1.72-1.95; W = 1.16, 1.04-1.28; H = 1.54, 1.43-1.65), right adrenal gland (L = 0.38, 0.33-0.57; H = 0.19, 0.15-0.31) and left adrenal gland (L = 0.36, 0.32-0.39; H = 0.18, 0.17-0.20). All kidney measurements were positively correlated with animal weight ( P < 0.05) but had no significant correlation to age. Measurements did not have any significant relationship to evaluated blood and urine parameters. Results from this study establish baseline measurements for callimico kidneys and adrenal glands to help clinicians use these imaging modalities for evaluation of these organs in this endangered species.

TRANSMISSION OF PTERYGODERMATITES NYCTICEBI IN A COLONY OF GOELDI'S MONKEYS ( CALLIMICO GOELDII) AND EVALUATION OF TREATMENT AND CONTROL.

Over a 2-yr period, four Goeldi's monkeys ( Callimico goeldii) died in a private zoo due to infections with the spirurid nematode Pterygodermatites nycticebi. Therapeutic measures with different anthelmintics were not successful. Due to the severe consequences caused by these infections, different actions were initiated, including sanitation measures and controlling of potential intermediate hosts (coprophagous arthropods). To identify possible intermediate hosts, arthropod species detected in the enclosure-parasite-free German cockroaches ( Blattella germanica), European earwigs ( Forficula auricularia), and rough woodlice ( Porcellio scaber)-were experimentally fed with feces of monkeys with patent P. nycticebi infections, resulting in established infections with third-stage larvae (L3) in roaches and earwigs. Furthermore, spiruroid L3 were detectable in 43% of the roaches and 30% of earwigs caught at the zoo. Polymerase chain reaction and sequence analysis of eggs, larval, and adult stages resulted in identical results, confirming the establishment of the parasite's life cycle in the zoo. This is the first documentation of the vector capacity of the European earwigs for P. nycticebi. As a measure of sanitation, a large part of the enclosure was emptied and cleaned. The Goeldi's monkeys were quarantined and treated with levamisole (7.5 mg/kg sc twice in intervals of 2 wk). Repeated coprologic examinations by zinc chloride flotation were undertaken. After the levamisole therapy, eggs were not found in the feces for 3 mo. However, shortly after resettling the monkeys into the sanitized enclosure, reshedding of small amounts of spirurid eggs was observed, whereupon deworming with levamisole was prescribed several times per year. The sanitation measures and the elimination of the intermediate hosts in a natural enclosure are presented as an example of the long-term controlling of the parasites.

Chromosome painting of the red-handed tamarin (Saguinus midas) compared to other Callitrichinae monkeys.

Here we present, for the first time, the complete chromosome painting map of Saguinus midas, the red-handed tamarin. Chromosome banding and painting with human chromosome-specific probes were used to compare the karyotype of this species with those of four other Neotropical primates of the subfamily Callitrichinae: Leontopithecus rosalia, Callithrix geoffroyi, C. penicillata, and Mico argentatus. The chromosome painting map of S. midas was identical to that of L. rosalia and other previously studied tamarin species (genera Saguinus and Leontopithecus). The three marmoset species studied (genera Callithrix and Mico) differed in the painting pattern of four human probes (chromosomes 1, 2, 10, and 16). These paints identified the presence or absence of chromosome associations HSA 1/10 and 2/16 in these taxa. By integrating our data with those from the literature, we were able to propose an ancestral Callitrichinae karyotype. The genera Saguinus and Leontopithecus (tamarins) conserve the ancestral Callitrichinae karyotype, while Mico and Callithrix (marmosets) show more derived karyotypes due to chromosome translocations and fissions that occurred during the evolution of these taxa.

Collection, Evaluation, and Coagulum Dissolution of Semen from Goeldi's Monkey, Callimico goeldii.

Research on Neotropical primates' reproduction is necessary due to the lack of available information and the increasing threat to these species. Callimico goeldii is listed as Vulnerable on the IUCN Red List of Threatened Species. This study aimed to test rectal electrostimulation for semen collection and evaluate seminal characteristics. Therefore, semen from 6 captive Goeldi's monkeys was collected and, for the first time, seminal characteristics are described. Coagulum formation was noted in all ejaculates, and we obtained partial or complete liquefaction of the samples. Results were (means ± SD): volume = 26.9 ± 11.87 µL; pH = 7.61 ± 0.28; concentration = 143.18 ± 174.96 × 106 spermatozoa/mL; total sperm motility = 83.33 ± 5.16%; linear progressive motility = 46 ± 24.08%; plasma membrane integrity = 36.38 ± 16.11%; acrosome integrity using fast-green/bengal-rose staining = 63.41 ± 11.72%, and kit Spermac® = 69.36 ± 11.81%; abnormal sperm = 72.5 ± 17.7%, with 16.2 ± 7.7% major defects and 56.3 ± 10% minor defects; sperm with high mitochondrial activity class I = 16.45 ± 22.25%. Rectal electrostimulation was an efficient method for semen collection in this species. Investigations are required to improve semen collection and handling, including cryopreservation methods.

Comparison of 3 Methods for Preventing Perianesthetic Hypothermia in Callimicos (Callimico goeldii).

Perianesthetic hypothermia is one of the most common complications in veterinary anesthesia, especially in small patients with a large body surface area to mass ratio. During anesthesia, body heat can be lost through 4 mechanisms-radiation, convection, conduction, and evaporation-but anesthetists frequently address only one mechanism at a time. Here we sought to evaluate 3 methods of preventing perianesthetic hypothermia in callimicos (Callimico goeldii). In our experience, these small NHP routinely become hypothermic under even brief inhalant anesthesia. To address multiple routes of heat loss, animals received 1 of 3 treatments: 1) placement of a reflective blanket over the patient to limit radiative heat loss to the surrounding environment; 2) placement of a reflective blanket and use of a heated anesthetic circuit, which warmed the inspired air to 104 °F (40 °C), and 3) placement under the patient of a forced-air warming blanket set at 109.4 °F (43 °C). Sources of radiative heat loss were assessed by using infrared thermography. Each animal was anesthetized with isoflurane and maintained in sternal recumbency in a temperature-controlled room (65 °F; 18.3 °C); esophageal core body temperature was monitored every 5 min for a total of 30 min. The rate of heat loss did not differ between the use of a reflective blanket with or without a heated anesthetic circuit. Animals provided the forced-air warming blanket experienced a slight increase in average body temperature. According to these findings, an underbody warm-air blanket provided the best protection against hypothermia for callimicos in sternal recumbency.

Hepatic myelolipoma in two Goeldi's monkeys from South America held captive.

Two cases of hepatic myelolipoma in Goeldi's monkeys from South America are described. One was a female evaluated due progressive abdominal distension. Ultrasound and computed tomography detected hepatic mass. Partial hepatectomy was performed, but the monkey died after surgery. Case 2 was a male that died suddenly, showing non-specific clinical signs.

EVALUATION OF IN-HOUSE URINE DIPSTICK, REFERENCE LABORATORY URINALYSIS, AND URINE PROTEIN: CREATININE RATIO FROM A COLONY OF CALLIMICOS (CALLIMICO GOELDII).

This study evaluated results from an in-house (IH) laboratory dipstick, a reference laboratory (RL) urinalysis, and urine protein : creatinine (UPC) ratios from callimicos ( Callimico goeldii ). Urine was collected from 25 individuals comprising a single colony under professional care in North America and compared based on laboratory, sex, age class, and presence or absence of a normal urinalysis. Urine specific gravity and pH between laboratories were statistically different. Overall, 56% to 100% of animals had at least a trace amount of protein in their urine. In comparing normal and abnormal urinalyses, IH dipstick protein, RL dipstick protein, RL pH, quantitative protein measurement, and UPC ratios were all statistically different. Eleven animals (44%) had UPC ratios that were ≥0.5. Based on results of this study, UPC > 0.3 was found to be abnormal and supportive of renal compromise in callimicos. Higher protein concentrations on the IH dipstick, the quantitative protein concentration, and UPC ratio in the 1- to 4-yr-old age class were the only significant age-related differences. There was no association between any categorical variable (glucose, blood, bilirubin, ketones, urobilinogen) and abnormal urinalysis. There were no differences between sexes. Since renal disease is a major cause of morbidity and mortality in this species, the authors recommend including routine urinalyses and UPC ratios as part of preventive care programs for callimicos. These data provide the first published information on urinalysis and UPC ratios in callimicos and will serve as a helpful reference for interpreting results and evaluating patients with renal disease.

Environmental complexity and feeding enrichment can mitigate effects of space constraints in captive callitrichids.

Non-human primates housed in zoos and laboratories often exhibit reduced activity and this poses welfare concerns. We examined the effects of enclosure types of differing size and environmental complexity on the activities of two species of callitrichids. We found that cotton-top tamarins housed in an enclosure of larger size and more environmental complexity showed higher activity levels, which was mainly contributed by more feeding/foraging activity. By contrast, Goeldi's monkeys housed in an enclosure of larger size and more environmental complexity showed lower activity levels, which was mainly contributed by less locomotory activity. In both species, off-exhibit groups housed in smaller enclosures did not show significantly less locomotory activity which would have been expected, as larger availability spaces should allow more opportunities for locomotion. Furthermore, the feeding enrichment had significant effects on increased feeding/foraging activity for both cotton-top tamarins and Goeldi's monkeys, irrespective of enclosure type. These results suggested that environmental complexity (or application of feeding enrichment) that provided more opportunities for natural foraging could have a larger effect on overall activity levels compared with larger enclosure sizes that should provide more locomotion opportunities. More importantly, it showed that even when enclosure space and complexity were limited, increased opportunities for foraging through the application of enrichment could increase species-typical behaviours. Such inexpensive, easy to implement enrichment methods should be applied to provide more complex environments for captive non-human primates, particularly in situations where there are logistical and/or cost constraints to the modification of physical exhibits.

INFECTION OF A GOELDI'S MONKEY (CALLIMICO GOELDII) WITH A EUROPEAN STRAIN OF ECHINOCOCCUS MULTILOCULARIS IN A CANADIAN INSTITUTION.

A 12-yr-old female Goeldi's monkey (Callimico goeldii) in British Columbia, Canada was diagnosed with alveolar echinococcosis (AE) on postmortem examination. Echinococcus multilocularis has been identified in several species of nonhuman primates, most frequently Old World primates, in zoos and research facilities in Europe and Asia. The strain affecting this monkey was identified as a European haplotype, indistinguishable from E. multilocularis recently identified in several canids in British Columbia. The animal is suspected to have been exposed while living in a zoological institution in Alberta, where E. multilocularis has also been reported in urban coyotes. Alveolar echinococcosis is a zoonotic disease of increasing concern in the United States and Canada, and this disease should be considered on the differential list of any nonhuman primate exhibiting signs of abdominal pain or distension, along with diagnostic imaging consistent with cystic structures of the liver or other organs.

RETROSPECTIVE REVIEW OF THE PREVALENCE OF MYELOLIPOMA IN GOELDI'S MONKEYS (CALLIMICO GOELDII).

Myelolipomas are mesenchymal tumors composed of adipose and hematopoietic tissue. They have been reported in many species, including domestic dogs and cats, birds, mustelids, wild felids, and nonhuman primates. Myelolipomas in Callitrichidae have been reported at postmortem examination and rarely antemortem. Multiple cases of hepatic myelolipomas associated with morbidity in Chicago Zoological Society's collection of Goeldi's monkeys (Callimico goeldii) prompted a global retrospective study to determine the prevalence and investigate factors associated with this condition. A total of 842 postmortem examination reports (1965-2013) collected from 133 captive collections were reviewed. Myelolipomas were reported in 17.2% (n=145) of animals, with significantly more female cases than male (P=0.023). There was a significantly older mean age at death in affected animals (134 months) compared to unaffected animals (79 months) (P<0.0001). Myelolipomas were diagnosed in the liver in all affected animals, and rarely in the adrenal or spleen. One in five (22.1%) affected Callimico had notable morbidity associated with this disease, which is markedly different from the primarily benign reports in other species. To identify disease, prevent morbidity from hepatic myelolipoma, and improve care of Callimico species in zoological collections, new Species Survival Plan (SSP) medical management recommendations include incorporating abdominal ultrasonographic examination during routine physical examinations in addition to previously recommended hematological and serum biochemical evaluations. Future studies are merited to examine the prevalence, risk factors, health effects, and treatment options of myelolipoma in living collections.

Lost and found: The third molars of Callimico goeldii and the evolution of the callitrichine postcanine dentition.

This study tests the hypothesis that the third molars of Callimico goeldii represent a reversal in evolutionary tooth loss within the Callitrichinae. Loss of third molars is part of a suite of unusual characters that has been used to unite marmosets and tamarins in a clade to the exclusion of Callimico. However, molecular phylogenetic studies provide consistent support for the hypothesis that marmosets are more closely related to Callimico than to tamarins, raising the possibility that some or all of the features shared by marmosets and tamarins are homoplastic. Here, I use the binary-state speciation and extinction (BiSSE) model and a sample of 249 extant primate species to demonstrate that, given the shape of the primate phylogenetic tree and the distribution of character states in extant taxa, models in which M3 loss is constrained to be irreversible are much less likely than models in which reversals are allowed to occur. This result provides support for the idea that the last common ancestor of Callimico and marmosets was characterized by the two-molared phenotype. The M3s of Callimico therefore appear to be secondarily derived rather than plesiomorphic. This conclusion may also apply to the other apparently plesiomorphic traits found in Callimico. Hypotheses regarding the re-evolution of M3 in the callitrichine clade and the origin and maintenance of the two-molared phenotype are discussed.

Determination of vertebral heart score in Goeldi's monkeys (Callimico goeldii).

BACKGROUND: Goeldi's monkeys (Callimico goeldii) are callitrichid primates commonly kept in zoological collections, and to date, no cardiac parameters have been recorded. The vertebral heart score (VHS) is an objective method of evaluation of cardiac size well documented in domestic mammals, and the aim of this study was to determine the VHS in Goeldi's monkeys. METHODS: In this retrospective study, right lateral radiographs of thirteen clinically well animals were reviewed and vertebral heart score determined. RESULTS: The vertebral heart score was found to be 9.35 ± 0.31. CONCLUSIONS: The observed value appears consistent within the study population and with values for other primate species. The value determined may be of benefit in objectively evaluating cardiac size in this species.

Isolation and characterization of chitin-degrading micro-organisms from the faeces of Goeldi's monkey, Callimico goeldii.

AIMS: The objective of this study was the isolation and characterization of chitin-degrading micro-organisms from the faeces of the insectivorous Goeldi's monkey, Callimico goeldii. METHODS AND RESULTS: Faeces samples were screened for chitin-degrading bacteria using basal medium in which chitin was included as the carbon and energy source. Of fifteen bacterial isolates with chitin-degrading activity, fourteen were also capable of degrading cellulose. All isolates were either aerobes or facultative anaerobes. CONCLUSIONS: Phylogenetic analyses of those isolates exhibiting strongest activity, as determined by the most distinctive zones of clearing in chitin-supplemented medium, were identified as Cellulosimicrobium spp., Arthrobacter spp., Staphylococcus spp. and Enterobacteriaceae. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports on the isolation of chitin-degrading microflora from nonhuman primates. Considering that chitin and cellulose are the most abundant naturally occurring polymers, it is of interest to note that the majority of isolates are capable of digesting both substrates. This may be of significance given that omnivorous primates live in seasonal environments, where the availability of food items varies with the seasons. Furthermore, given the presence of a chitin-degrading microflora, this may have implications, in terms of the inclusion of fungi and/or insects in the diets of these animals in captivity, whether as part of medical research or conservation programmes.