Green rooibos extract (GRE), shown to improve hyperglycemia and HDL/LDL blood cholesterol, has potential as a nutraceutical beverage ingredient. The main bioactive compound of the extract is aspalathin, a C-glucosyl dihydrochalcone. The study aimed to determine the effect of common iced tea ingredients (citric acid, ascorbic acid, and xylitol) on the stability of GRE, microencapsulated with inulin for production of a powdered beverage. The stability of the powder mixtures stored in semi-permeable (5 months) and impermeable (12 months) single-serve packaging at 30 °C and 40 °C/65% relative humidity was assessed. More pronounced clumping and darkening of the powders, in combination with higher first order reaction rate constants for dihydrochalcone degradation, indicated the negative effect of higher storage temperature and an increase in moisture content when stored in the semi-permeable packaging. These changes were further increased by the addition of crystalline ingredients, especially citric acid monohydrate. The sensory profile of the powders (reconstituted to beverage strength iced tea solutions) changed with storage from a predominant green-vegetal aroma to a fruity-sweet aroma, especially when stored at 40 °C/65% RH in the semi-permeable packaging. The change in the sensory profile of the powder mixtures could be attributed to a decrease in volatile compounds such as 2-hexenal, (Z)-2-heptenal, (E)-2-octenal, (E)-2-nonenal, (E,Z)-2,6-nonadienal and (E)-2-decenal associated with “green-like” aromas, rather than an increase in fruity and sweet aroma-impact compounds. Green rooibos extract powders would require storage at temperatures ≤ 30 °C and protection against moisture uptake to be chemically and physically shelf-stable and maintain their sensory profiles.

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.

Scent is known to play an important role in the reproduction of cheetahs and other felids. In fact, the presence/odor of a male cheetah has been noted to trigger the estrous cycle in females. The objective of this study was to analyze the marking fluid (MF) of male cheetahs from different breeding groups to determine the composition of volatile organic compounds (VOCs) present, with the aim of identifying potential pheromones relating to sexual behavior/attraction in this species. Four breeding (B; age: 8.9 ± 1.3 years old) and four non-breeding (NB; age: 5.5 ± 0.8 years old) males were selected for this study. Samples were collected into a glass beaker, transferred immediately into a 20 mL glass screw-cap vial with a polytetrafluoroethylene (PTFE) coated silicone septum, and stored until analyzed by headspace solid-phase microextraction (HS-SPME) using gas chromatography–mass spectrometry. A contingency test with Fisher’s exact test, using the frequency (FREQ) procedure of SAS 9.4, was conducted to determine the difference between the number of VOCs identified per breeding group; furthermore, differences in relative concentration (RC) of the identified VOCs between breeding groups were analyzed using ANOVA for repeated measures with the GLIMMIX procedure. From the 13 MF samples analyzed, 53 VOCs were identified, and 12 were identified in all the samples. Five of these (dimethyl disulfide, benzaldehyde, acetophenone, phenol, and indole) are known to be involved in attraction/sexual behavior in mammals. Between the two groups, the RC of indole was significantly higher in the NB group, whereas the RC of dodecanoic acid was significantly higher in the B group. Although not significant, the RC of benzaldehyde was higher in the B versus the NB group. The results of this study do support the hypothesis of differences in VOCs’ between B and NB male cheetahs. However, the overlapping of age and breeding status and the diet differences could not be controlled. Still, the evidence of changes in MF composition in male cheetahs necessitates further studies on possible strategies to improve reproduction in captivity.