Age is a key demographic in conservation where age classes show differences in important population metrics such as morbidity and mortality. Several traits, including reproductive potential, also show senescence with ageing. Thus, the ability to estimate age of individuals in a population is critical in understanding the current structure as well as their future fitness. Many methods exist to determine age in wildlife, with most using morphological features that show inherent variability with age. These methods require significant expertise and become less accurate in adult age classes, often the most critical groups to model. Molecular methods have been applied to measuring key population attributes, and more recently epigenetic attributes such as methylation have been explored as biomarkers for age. There are, however, several factors such as permits, sample sovereignty, and costs that may preclude the use of extant methods in a conservation context. This study explored the utility of measuring age-related changes in methylation in candidate genes using mass array technology. Novel methods are described for using gene orthologues to identify and assay regions for differential methylation. To illustrate the potential application, African cheetah was used as a case study. Correlation analyses identified six methylation sites with an age relationship, used to develop a model with sufficient predictive power for most conservation contexts. This model was more accurate than previous attempts using PCR and performed similarly to candidate gene studies in other mammal species. Mass array presents an accurate and cost-effective method for age estimation in wildlife of conservation concern.
Ouderdom is ‘n sluitsteen kenmerk in dier bewaring waar ouderdoms klasse verskille toon in belangrike populasie maatstawwe soos morbiditeit en mortaliteit. Verskeie eienskappe, soos voortplantings potensiaal, toon ook aansienlike verandering met veroudering. Dus is die vermoë om ouderdom te bepaal vir individue binne ‘n populasie krities tot die ontleding van huidige struktuur en toekomstige welstand. Verskeie metodes bestaan reeds vir ouderdom bepaling waarvan meeste gebruik maak van morfologiese kenmerke wat voorspelbare verandering toon met ouderdom. Hierdie metode vereis egter merkwaardige kennis en word minder akkuraat in volwassene klasse wat die mees krities is om te bepaal. Molekulêre metodes word reeds aangewend om kern populasie eienskappe te bepaal en meer onlangs is epigenetiese eienskappe soos metilering ondersoek as ‘n biologiese merker vir ouderdom. Daar is egter verskeie faktore soos permitte, monster eienaarskap, en kostes wat die toepassing van huidige metilering metodes in ‘n bewaring konteks mag inperk. Die huidige studie ondersoek die gebruiklikheid van ouderdom verwante verandering in metilering, soos gemeet deur massa‐skikking tegnologie, in kandidaat gene. Nuwe metodiek word beskryf vir die gebruik van ortologiese gene om teikens te identifiseer en skerm vir differensiële metilering. Om die spesifieke toepassing te illustreer is die Afrika jagluiperd as gevallestudie gebruik. Korrelasie analise het ses gemetileerde teikens ïdentifiseer met ‘n ouderdoms verwante profiel wat ingesluit is in die ontwikkeling van ‘n model vir ouderdom bepaling met genoegsame akkuraatheid vir meeste bewaring kontekste. Die model was meer akkuraat as voormalige proewe met PKR en het soortgelyk gevaar as ander kandidaat geen studies in verskeie soogdiere. Massa‐skikking bied dus ‘n akkurate en bekostigbare metode om ouderdom te bepaal in wilde diere van belang vir bewaring.

Unnatural diet composition and frequent feeding regimes may play an aetiological role in the multiple diseases prevalent in captive cheetahs. This study investigated the responses of captive-born (hand-reared) cheetahs (n = 6) to a reduced feeding frequency schedule distinguished by offering larger quantities of food less frequently. The study cheetahs were fed four once-daily meals per week during the 3-week treatment period, followed by a 3-week control period in which they were fed two daily rations six days a week. Total weekly food intake was maintained throughout the study. Variations in behaviour, faecal consistency score (FCS), and faecal glucocorticoid metabolite concentration were measured. Less frequent feeding resulted in higher FCS (p < 0.01) and locomotory behaviour (p < 0.05) among the studied cheetahs. Faecal glucocorticoid metabolite concentration demonstrated an initial acute stress response to the change in feeding frequency (p < 0.05) and subsequent adaptation. The results of the FCS analysis suggest that the more natural feeding pattern could have benefited the studied cheetahs\' gastrointestinal health without a significant behavioural or physiological stress response overall to the change in feeding frequency.

Herein, we describe the management of nasopharyngitis caused by Schizophyllum commune infection in a captive cheetah. Computed tomography revealed a nodule in the nasal cavity and pharynx, and an endoscopic biopsy was performed. As a result, the nodule was surgically resected because of a suspected carcinoma. However, the surgical specimen was histologically re-evaluated and a fungal granuloma was diagnosed. Sequence analysis of DNA from formalin-fixed, paraffin-embedded samples revealed S. commune infection. The cheetah was administered fluconazole orally for 73 days. However, the drug was ineffective and itraconazole was administered for 14 days. Symptoms such as nasal discharge and sneezing have completely resolved for 4 years.

Smaller predators may overcome body size restrictions on their prey base by selecting for juveniles of larger prey species. However, traditional prey selection models ignore demographic classes within prey species. We refined these models for two predators with contrasting body sizes and hunting strategies, by including seasonal consumption and availability of prey demographic classes. We predicted that cheetahs would select for smaller neonate and juvenile prey especially of larger species, while lions would select for larger, adult prey. We further predicted seasonal diet shifts in cheetah, but not lion. We recorded species-specific demographic class prey use (kills) via direct observation and GPS cluster of cheetahs and lions fitted with GPS collars. Species-specific demographic class prey availability was estimated from monthly driven transects, and species-specific demographic class prey preferences were estimated. The availability of prey demographic classes varied seasonally. Cheetahs preferred neonates, juveniles, and sub-adults during the wet season, but adults and juveniles during the dry season. Lions preferred adult prey irrespective of season, with sub-adults, juveniles, and neonates killed relative to their abundance. This confirms that traditional prey preference models do not adequately account for demographic-specific prey preference. This is particularly important for smaller predators, like cheetahs, that focus on smaller prey but can expand their prey base by killing juveniles of larger species. For these smaller predators, prey availability will vary strongly seasonally, making them more vulnerable to processes that influence prey reproduction, like global change.

In cheetahs, age at first parturition correlates negatively with reproductive lifespan (asymmetric reproductive aging); therefore, breeding cheetahs at a young age is essential to maximize reproductive performance in this species. However, younger females display a significantly reduced frequency of copulatory behaviour, which negatively affects breeding. Volatile organic compounds (VOCs) are known to regulate appropriate behavioural responses in various species, including reproductive behaviour; moreover, they have proven to play a role in captive breeding methods in cheetahs, as well as mate choice. Therefore, the objective of this study was to evaluate the effect of a synthetic scent (SS) on the frequency of the five oestrous behaviour(s) (sniff, rub, roll, spray, and meow-chirp) known to be indicative of oestrus in female cheetahs. Based on the results of a previous study from our research group, five VOCs, identified in the marking fluid of male cheetahs, and known to be pheromones involved in reproductive behaviour, were used to create the SS. This was accomplished by mixing benzaldehyde, acetophenone, indole, dimethyl disulphide and phenol with (99.9%) ethanol. Seven female cheetahs were then observed for one oestrus cycle without stimulation (control) and then once again while exposed to the SS (treatment), which was sprayed on foil trays placed around the outside of each enclosure. The occurrence of the five oestrous behaviours was recorded and tallied per day of observations. Although the SS did not have a significant effect on the frequency of oestrous behaviours displayed by the females used in this study, five of the seven (71%) did show an increase in their behaviour with the SS when oestrogen concentrations were at their highest (peak oestrus), including three of the four younger females. The SS also significantly increased the sniffing behaviour in general. Although the results of this study do indicate that VOCs influence cheetahs and their behaviour, firm conclusions cannot be drawn due to the low number of animals used, as well as the significant effect the observation methods had on the results. Nonetheless, this study represents the first of this kind in cheetahs, therefore representing an important step in determining the role of VOCs in aiding breeding in captivity.

A 3.5-year-old female cheetah (Acinonyx jubatus) died after a 10-day history of anorexia, regurgitation and diarrhoea despite symptomatic therapy. At gross post-mortem examination, the stomach was blood-filled with mucosal thickening and multifocal ulcerations. The intestinal mucosa was thickened and reddened, and the intestinal lumen was filled with dark red to black pasty content. Gastric histological lesions were compatible with gastritis due to Helicobacter infection, which was confirmed by polymerase chain reaction. Histology of the intestines revealed a severe necrotizing neutrophilic enterocolitis with abundant intralesional curved to spiral bacteria, corresponding to Campylobacter jejuni, which were subsequently isolated from both small and large intestinal contents. No other intestinal pathogens were detected despite thorough investigations. These findings suggest that C. jejuni may have played an aetiological role in the enterocolitis. Such an association has not been previously reported in non-domestic felids.

OBJECTIVE: To evaluate the agreement between high-definition oscillometry (HDO) used on the metatarsus or tail base with invasive arterial blood pressures measured in the dorsal pedal artery in anaesthetised cheetahs.
METHODS: Prospective clinical study.
METHODS: A group of 13 captive adult cheetahs.
METHODS: Cheetahs were immobilised with medetomidine (32-45 μg kg-1) and tiletamine/zolazepam (0.93-1.39 mg kg-1) administered intramuscularly, and anaesthesia was maintained with either isoflurane in oxygen or continuous propofol infusion. Invasive blood pressure was measured via a 20 gauge intra-arterial catheter in the dorsal pedal artery in the metatarsus and used as a reference method for pressures simultaneously estimated using HDO on the contralateral metatarsus and tail base. Bland-Altman plots (for repeated measurements) and criteria defined by the American College of Veterinary Internal Medicine (ACVIM) were used to compare agreement according to the anatomical location of the cuff, the anaesthetic maintenance agent and magnitude of the blood pressure.
RESULTS: A total of 147 paired measurements were obtained with HDO on the metatarsus and 135 on the tail. Agreement with invasive pressures was better when HDO was used on the tail (rather than on the metatarsus) with all ACVIM criteria being met. Mean bias (a positive bias meaning that HDO overestimated the invasively measured pressures) ± standard deviation of differences for systolic, diastolic and mean arterial pressures were -7.0 ± 13.9, 4.2 ±12.1 and 4.6 ±11.2 mmHg, respectively, for HDO on the tail, and -11.9 ±15.1, 2.8 ±16.5 and 2.1 ±13.2 mmHg, respectively, for HDO on the metatarsus. Agreement was better during isoflurane anaesthesia than propofol, and at lower blood pressures than at higher.
CONCLUSIONS: When used on the tail base of anaesthetised cheetahs, HDO met the ACVIM validation criteria for a noninvasive device, as compared to invasively measured pressures in the dorsal pedal artery.

Fluoroquinolones are a widely used class of chemotherapeutics within veterinary medicine, prized for their broad-spectrum bactericidal activity. These drugs present a known risk of retinal phototoxicity in domestic cats (Felis catus); therefore, using lower doses and alternative antibiotic classes is encouraged in this species. This adverse drug effect of fluoroquinolones, and enrofloxacin specifically, has been determined to be species-specific in domestic felids. Four feline-specific missense variants in ABCG2 result in four amino acid changes (E159M, S279L, H283Q, and T644I) that are unique to the domestic cat compared with multiple other nonfeline mammalian species. These changes alter the ABCG2 protein involved with the cellular transmembrane transport of drugs, including fluoroquinolones, making the protein functionally defective in domestic cats. The predisposition to fluoroquinolone-mediated phototoxicity in nondomestic felids was explored in this study. At least eight nondomestic felids share the four ABCG2 missense variants with domestic cats, and eleven other felids shared at least three of the four domestic cat variants. Taken together, these results suggest the genetic potential for nondomestic felids to also experience fluoroquinolone-induced retinal phototoxicity; therefore, cautions similar to those for domestic cats should be followed for these drugs in the entire feline taxon.

African large carnivores have undergone significant range and population declines over recent decades. Although conservation planning and the management of threatened species requires accurate assessments of population status and monitoring of trends, there is evidence that biodiversity monitoring may not be evenly distributed or occurring where most needed. Here, we provide the first systematic review of African large carnivore population assessments published over the last two decades (2000-2020), to investigate trends in research effort and identify knowledge gaps. We used generalised linear models (GLMs) and generalised linear mixed models (GLMMs) to identify taxonomic and geographical biases, and investigated biases associated with land use type and author nationality. Research effort was significantly biased towards lion (Panthera leo) and against striped hyaena (Hyaena hyaena), despite the latter being the species with the widest continental range. African wild dog (Lycaon pictus) also exhibited a negative bias in research attention, although this was partly explained by its relatively restricted distribution. The number of country assessments for a species was significantly positively associated with its geographic range in that country. Population assessments were biased towards southern and eastern Africa, particularly South Africa and Kenya. Northern, western, and central Africa were generally under-represented. Most studies were carried out in photographic tourism protected areas under government management, while non-protected and trophy hunting areas received less attention. Outside South Africa, almost half of studies (41%) did not include authors from the study country, suggesting that significant opportunities exist for capacity building in range states. Overall, large parts of Africa remain under-represented in the literature, and opportunities exist for further research on most species and in most countries. We develop recommendations for actions aimed at overcoming the identified biases and provide researchers, practitioners, and policymakers with priorities to help inform future research and monitoring agendas.

Fecal proteomics allows for the identification of proteins and peptides present in stools and is useful in finding possible new biomarkers for diagnosing and/or monitoring gastrointestinal (GI) disorders. In the present study, we investigated the fecal proteome in healthy and diseased cheetahs (Acinonyx jubatus). Captive individuals of this species frequently show gastrointestinal disorders characterized by recurrent episodes of diarrhea, rare episodes of vomiting and weight loss, associated with Helicobacter spp. infection. Fecal proteomic evaluation has been performed by two-dimensional electrophoresis followed by liquid chromatography-tandem mass spectrometry. In healthy cheetahs, the results showed the presence of the following proteins: collagen alpha-1 (II) chain, transthyretin, IgG Fc-binding protein, titin, dystonin, isopentenyl-diphosphate Delta-isomerase 1, sodium/potassium-transporting ATPase subunit alpha-1 and protein disulfide-isomerase A6. The presence of albumin isoforms was found only in diseased cheetahs. The present paper reports the study of the fecal proteome in the cheetah, evidences some differences between healthy and diseased patients and confirms, once again, the potential of fecal proteomics for the study of the GI environment, with promising developments regarding the identification of new diagnostic/monitoring markers.