A method for noninvasive individual genotyping of black-footed cat (Felis nigripes).

The black-footed cat (Felis nigripes) is endemic to the arid regions of southern Africa. One of the world's smallest wild felids, the species occurs at low densities and is secretive and elusive, which makes ecological studies difficult. Genetic data could provide key information such as estimates on population size, sex ratios, and genetic diversity. In this study, we test if microsatellite loci can be successfully amplified from scat samples that could be noninvasively collected from the field. Using 21 blood and scat samples collected from the same individuals, we statistically tested whether nine microsatellites previously designed for use in domestic cats can be used to identify individual black-footed cats. Genotypes recovered from blood and scat samples were compared to assess loss of heterozygosity, allele dropout, and false alleles resulting from DNA degradation or PCR inhibitors present in scat samples. The microsatellite markers were also used to identify individuals from scats collected in the field that were not linked to any blood samples. All nine microsatellites used in this study were amplified successfully and were polymorphic. Microsatellite loci were found to have sufficient discriminatory power to distinguish individuals and identify clones. In conclusion, these molecular markers can be used to monitor populations of wild black-footed cats noninvasively. The genetic data will be able to contribute important information that may be used to guide future conservation initiatives.

The potential and shortcomings of mitochondrial DNA analysis for cheetah conservation management.

There are only about 7,100 adolescent and adult cheetahs (Acinonyx jubatus) remaining in the wild. With the majority occurring outside protected areas, their numbers are rapidly declining. Evidence-based conservation measures are essential for the survival of this species. Genetic data is routinely used to inform conservation strategies, e.g., by establishing conservation units (CU). A commonly used marker in conservation genetics is mitochondrial DNA (mtDNA). Here, we investigated the cheetah's phylogeography using a large-scale mtDNA data set to refine subspecies distributions and better assign individuals to CUs. Our dataset mostly consisted of historic samples to cover the cheetah's whole range as the species has been extinct in most of its former distribution. While our genetic data largely agree with geography-based subspecies assignments, several geographic regions show conflicting mtDNA signals. Our analyses support previous findings that evolutionary forces such as incomplete lineage sorting or mitochondrial capture likely confound the mitochondrial phylogeography of this species, especially in East and, to some extent, in Northeast Africa. We caution that subspecies assignments solely based on mtDNA should be treated carefully and argue for an additional standardized nuclear single nucleotide polymorphism (SNP) marker set for subspecies identification and monitoring. However, the detection of the A. j. soemmeringii specific haplogroup by a newly designed Amplification-Refractory Mutation System (ARMS) can already provide support for conservation measures. Supplementary Information: The online version contains supplementary material available at 10.1007/s10592-022-01483-1.

TREATMENT OF MYCOBACTERIOSIS CAUSED BY MYCOBACTERIUM AVIUM SSP. HOMINISSUIS IN A GROUP OF CAPTIVE LOWLAND TAPIRS (TAPIRUS TERRESTRIS).

Tapirs are a taxonomic group with a high susceptibility to mycobacterial diseases. However, successful therapy has only been documented sporadically. Here treatment of mycobacteriosis diagnosed in three, one male and two female, lowland tapirs (Tapirus terrestris) in a zoo in Germany is reported. Two of the animals showed chronic mild respiratory signs, and conventional therapy did not improve the condition. Culture of broncho-alveolar lavage (BAL) samples was positive for Mycobacterium avium ssp. hominissuis. Upon airway endoscopy, bronchial edema and increased mucus production were visible. Initially, all three infected tapirs received oral antimycobacterial therapy consisting of 5 mg/kg body weight isoniazid, 10 mg/kg rifampicin, and 10 mg/kg clarithromycin q24h. Based on therapeutic drug level monitoring, the doses of rifampicin were adjusted to 12 and 15 mg/kg in the females and the male, respectively. The treatment with all three drugs was continued for 11 mon. Six months into treatment, the clinical condition resolved, and repeated BAL samples of all three tapirs tested negative for mycobacteria by culture. Here the approach for a treatment protocol with minimal side effects suitable to control infections with nontuberculous mycobacteria in lowland tapirs is reported.

Sex-specific actuarial and reproductive senescence in zoo-housed tiger (Panthera tigris): The importance of sub-species for conservation.

A fifth of all known species are currently classified as threatened in the wild: the rate of biodiversity loss is rapid, continuous, and mostly due to anthropogenic activities. To slow down this decline, the accurate estimation of demographic parameters for threatened species is critical. With this aim, zoo institutions play an important role, giving access to data on zoo-housed animals, which aids researchers working on species life-history traits and intrinsic factors influencing the fitness of both sexes, such as age. While tigers (Panthera tigris) are particularly threatened in their natural environment, few of their demographic parameters have been determined because of their solitary and elusive nature as well as low population density. Using individual-based information for more than 9200 tigers (from 1938 to 2018) recorded in the International Tiger Studbook 2018, we aimed to determine sub-species and sex-specific variability of survival and reproductive parameters with age. No significant sex-difference in actuarial senescence (i.e., decline of survival probabilities with age) was observed but males tended to have a higher juvenile mortality and a faster senescence than females. Reproductive senescence (i.e., decline of reproductive parameters with age) was more pronounced in females than males. Moreover, we observed sub-species-specific variation in mortality and reproductive patterns, pointing out the necessity to consider them independently for conservation goals. Our findings can provide meaningful improvements to the husbandry of zoo-housed tigers, emphasizing the importance of adult breeding females of 7-9 years-old to control zoo-housed population size, but also providing accurate demographic estimates, crucial to set up effective conservation plans.

Lifespan decreases with proportion of sons in males but not females of zoo-housed tigers and lemurs.

Several studies have shown higher costs of rearing sons than daughters in mammals where males are larger than females. These studies typically focus on females by examining how the offspring sex ratio during a single reproductive event affected mothers' subsequent reproduction or survival probability. Here, we examine relationships between offspring sex ratio during single or multiple reproductive events and several survival metrics in mothers and fathers, using data from zoo-housed tigers (Panthera tigris) and ruffed lemurs (Varecia sp.). Our analyses failed to reveal an overall cost of reproduction or a higher cost of sons to mothers. In male ruffed lemurs, the proportion of sons produced during early life (before 10 years old) was negatively correlated with lifespan later in life. In tigers, males with a higher proportion of sons during their lifetime had shorter lifespans. One likely mechanism is the difference in testosterone levels between males: a high concentration of testosterone can increase the proportion of sons and compromise immune function. Our results suggest studies in wild populations should address the outstanding challenge of understanding consequences of sex allocation for males, and open an opportunity to predict lifespan in an applied conservation context.

Clinical outcome and diagnostic methods of atypical mycobacteriosis due to Mycobacterium avium ssp. hominissuis in a group of captive lowland tapirs (Tapirus terrestris).

Tapirs seem particularly susceptible to mycobacterial infections, especially to tuberculosis caused by M. tuberculosis or M. bovis. In this case series, we report an infection with the non-tuberculous mycobacteria (NTM) species M. avium ssp. hominissuis (MAH) in a group of four (2.2) captive lowland tapirs (Tapirus terrestris). Two female tapirs showed mild respiratory signs such as coughing and mucous sputum production for several years, one juvenile male tapir had to be euthanized due to severe dyspnoea, and the adult male only showed mild respiratory signs in 2010. Post-mortem histopathology of the euthanized animal revealed a chronic bronchopneumonia, and MAH was detected via culture. Subsequently, the three remaining tapirs were tested further: serologically, the tapirs had high antibody titres against M. avium, but they showed no reaction in the comparative skin test (TST). At several time points, the animals were tested for the presence of mycobacteria in different sample matrices including sputum samples, pooled faecal samples as well as swabs from the tapir enclosure to identify potential environmental niches of the pathogen. Moreover, animals were directly sampled using nasal swabs, endoscopic broncho-alveolar (BAL) and gastric lavages. MAH was detected by culture in the sputum samples, in the BAL of the breeding pair, as well as in the swimming pool water and walls, and in swabs taken from the tapir's sleeping beds. We conclude that the TST is not a useful diagnostic tool to detect MAC infections in tapirs, whereas antibody ELISA and culture from BAL appear more sensitive.

EVALUATION OF TWO DOSES OF BUTORPHANOL-MEDETOMIDINE-MIDAZOLAM FOR THE IMMOBILIZATION OF WILD VERSUS CAPTIVE BLACK-FOOTED CATS (FELIS NIGRIPES).

The efficacy, safety, physiologic effects, and reversibility of butorphanol-medetomidine-midazolam (BMM) immobilization were evaluated in black-footed cats (Felis nigripes) and compared between captive and wild animals. Nine captive and 14 wild black-footed cats were hand injected into an accessible hind limb muscle group with the BMM combination. The captive cats (captive group) received a lower dose of the combination (butorphanol, 0.25 ± 0.03 mg/kg; medetomidine, 0.06 ± 0.01 mg/kg; midazolam, 0.13 ± 0.02 mg/kg), whereas the wild cats received a higher dose (butorphanol, 0.53 ± 0.11 mg/kg, medetomidine, 0.13 ± 0.03 mg/kg, midazolam, 0.27 ± 0.05 mg/kg). Two capture methods were required to restrain the wild cats; previously collared cats were tracked and excavated out of their burrows during daylight hours (excavated group), whereas uncollared cats were randomly located using spotlights and pursued by a vehicle at night (pursued group). Inductions were rapid and no spontaneous arousals occurred. Mean arterial blood pressure in all cats was within normal limits for domestic cats. Initial rectal temperatures varied greatly among the groups, but decreased in all groups as the immobilization progressed. In the pursued animals, heart rates and respiratory rates were initially elevated. All cats had moderate hypoxemia, hypocapnia, and metabolic acidosis. Intramuscular administration of naltrexone, atipamezole, and flumazenil resulted in rapid, uncomplicated recoveries. BMM is thus a safe, effective immobilizing drug combination for both captive and wild black-footed cats, but higher doses are required in wild animals. The capture methods exerted a greater influence on the physiology of the immobilized animals than did the doses of the drugs used. Although this drug combination can be used safely to immobilize black-footed cats, supplemental oxygen should always be available for use, especially in pursued animals due to hypoxia.

Metabarcoding of eukaryotic parasite communities describes diverse parasite assemblages spanning the primate phylogeny.

Despite their ubiquity, in most cases little is known about the impact of eukaryotic parasites on their mammalian hosts. Comparative approaches provide a powerful method to investigate the impact of parasites on host ecology and evolution, though two issues are critical for such efforts: controlling for variation in methods of identifying parasites and incorporating heterogeneity in sampling effort across host species. To address these issues, there is a need for standardized methods to catalogue eukaryotic parasite diversity across broad phylogenetic host ranges. We demonstrate the feasibility of a metabarcoding approach for describing parasite communities by analysing faecal samples from 11 nonhuman primate species representing divergent lineages of the primate phylogeny and the full range of sampling effort (i.e. from no parasites reported in the literature to the best-studied primates). We detected a number of parasite families and regardless of prior sampling effort, metabarcoding of only ten faecal samples identified parasite families previously undescribed in each host (x̅ = 8.5 new families per species). We found more overlap between parasite families detected with metabarcoding and published literature when more research effort-measured as the number of publications-had been conducted on the host species' parasites. More closely related primates and those from the same continent had more similar parasite communities, highlighting the biological relevance of sampling even a small number of hosts. Collectively, results demonstrate that metabarcoding methods are sensitive and powerful enough to standardize studies of eukaryotic parasite communities across host species, providing essential new tools for macroecological studies of parasitism.

Morphological and physiological aspects of digestive processes in the graminivorous primate Theropithecus gelada-a preliminary study.

Hindgut fermentation has been suggested to contribute significantly to the digestive process in the gelada (Theropithecus gelada). We therefore hypothesized that in an in vitro fermentation test (Hohenheim gas test, using gas production as measure of microbial digestion) inoculum based on fresh gelada feces would degrade grass to a similar degree as zebra (Equus burchelli chapmani) feces and to a higher degree than that of hamadryas baboons (Papio hamadryas). Additionally, morphology of gelada tongue, salivary glands, stomach, and intestine were examined in this study. Gas production was measured between 4 and 96 hr using animal feces incubated with 200 mg of air-dry hay or mixed concentrate sample. For grass hay, 12-hr gas production was as follows: T. gelada (19.9 ml)>Papio (18.4 ml)>Equus (15.7 ml). After 24 hr, gas production changed: Papio (35.1 ml)>T. gelada (31.9 ml)>Equus (27.9 ml). At 96 hr, Papio was unexpectedly the most effective species with the highest gas production (53.1 ml)>zebra (51.2 ml)>gelada (49.4 ml). With a concentrate standard, 12-hr gas production was as follows: T. gelada (38.5 ml)>Equus (36.8 ml) = Papio (36.4 ml). At 24 hr, gas production differed: Papio (51.7 ml)>Equus (47.0 ml) = T. gelada (46.8 ml). At 96 hr, zebra was the most effective species with the highest gas production (63.9 ml)>Papio (60 ml) = T. gelada (59.9 ml). In conclusion, the results show that the microbial population present in gelada feces is able to ferment forage and concentrate substrates in vitro, although this fermentation did not occur with the expected effectiveness. Future studies should therefore focus also on the bacteria species involved.

Saliva of the graminivorous Theropithecus gelada lacks proline-rich proteins and tannin-binding capacity.

Gelada baboons are the sole survivors of the genus Theropithecus and the only known graminivorous primates. They developed special adaptations to their diet such as high-crowned teeth for processing hard and abrasive feed. The fine-tuning of salivary protein composition might be another key mechanism that is used by species for adapting to the environment and competing with rivals for exploiting new ecological niches. In order to test whether gelada (graminivorous) and hamadryas baboons (omnivorous) differ in their salivary protein composition, we compared whole saliva samples of captive Theropithecus gelada and Papio hamadryas using gel electrophoresis and tannin-binding assay. We hypothesized that the amount of proline-rich salivary proteins with tannin-binding capacity is higher in baboons consuming a feed with high dicot/monocot rations. Dicots produce tannins as a chemical defense system, discouraging animals from eating them. In contrast to dicots, monocots do not synthesize tannins. The presence of tannin-binding proteins in saliva should effectively inactivate the dicot tannin-based defense mechanism and increase the dietary breadth and/or the capability to switch between monocots and dicot leaves. The lack of such tannin-binding proteins in saliva would indicate a narrow dietary spectrum more restricted to monocots. We found T. gelada to completely lack proline-rich proteins (PRPs) and tannin-binding capacity similar to a great variety of other grazing mammals. In contrast, P. hamadryas does possess PRPs with tannin-binding activity. The findings support a growing body of evidence suggesting a high-level specialization of T. gelada to grass diets. However, it remains unclear, whether loss of salivary tannin-binding capacity drove the gelada into its narrow feeding niche, or whether this loss is the result of a long process of increased specialization. Thus, from an ecological point of view, T. gelada appears to be more vulnerable to environmental changes than other baboon species owing to its narrow dietary traits.