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Reports of the vaginal flora of wild cats such as lions or leopards are scarce. The micro-organisms most commonly found in the vagina of clinically healthy cats are aerobic bacteria such as coagulase-negative Staphylococcus, Streptococcus canis, and Escherichia coli. Simonsiella spp are large Gram-negative bacteria belonging to the Neisseriaceae family, typically found in the oral cavity and upper respiratory tract of many species. To date, there are no reports of the detection of Simonsiella spp in the vaginal flora of any felid. For a period of six months, daily behaviour monitoring was performed on six captive lionesses at a South African conservation centre, in parallel with the collection of vaginal swabs and interpretation of the resultant vaginal cytologies every other day. Oestrus was identified by typical female reproductive behaviours, as well as by enlarged and separated vulvar lips, and a predominant proportion of superficial cornified cells, clearing of the background, and high bacterial presence in the vaginal smear. Simonsiella spp were identified by their characteristic morphology in 58% (60 of 103) of the vaginal samples collected during oestrus. They were also found in oral swabs of three out of three lions tested. Additionally, Simonsiella spp were opportunistically found in a vaginal smear from a zoo housed female Sri Lankan leopard in oestrus, during a routine reproduction assessment. The finding of Simonsiella spp may be more common than previously suspected, transitory, and without detectable clinical relevance. A connection between occurrence of these bacteria and oestrus was apparent.

Vaginal cytology evaluation is an economic, non-invasive technique for indirect monitoring of fluctuations in estrogen concentrations, and thus progression of the estrous cycle. This technique is widely used in domestic dogs for determining timing of artificial insemination. There, however, are only a few reports on the vaginal cytology of non-domestic felids, including lions. This study was conducted, therefore, to describe the vaginal epithelial changes throughout the reproductive cycle of African lions, and to investigate the efficacy of vaginal cytology assessments for predicting reproductive stages. During a 12-month period, reproductive behavioral data and vaginal swabs were collected daily from five lionesses. In total, 541 vaginal smears were evaluated for the proportion of mucosal epithelial cells, neutrophils, bacterial cells, and amount of mucous, cellular debris. One single swab with a large proportion of superficial cells, absence of neutrophils, large number of bacteria, without cellular debris was sufficient for detecting lionesses in estrus. Likewise, one cytology sample with a large proportion of parabasal and intermediate cells, few neutrophils, few bacteria, and large amount of mucous, cellular debris enabled detection of females in advanced diestrus or gestation. To distinguish lionesses in early diestrus from those in an inter-estrous period, at least two consecutive swabs were necessary for satisfactory classification. Overall, evaluation of vaginal cytology samples was an effective technique for differentiation among different stages of the reproductive cycle, confirmation of estrus, and pregnancy diagnosis in lionesses. This technique, therefore, has the potential for application in classifying different stages of the reproductive cycle in other feline species.

Despite postulated potential for wildlife conservation, success of assisted reproduction techniques (ART) in ex-situ feline breeding remains25%. The aim of this project was to develop a simplified, non-surgical artificial insemination (AI) protocol for African lions (Panthera leo), using an exogenous GnRH analogue to induce ovulation in females presenting natural oestrus, and minimizing manipulation of the animals. Four protocols were tested in five trained lionesses (3.5-8 years), for a total of 14 inseminations (2-4 per lioness). These protocols differed in the time lapse between GnRH injection and insemination, on days 4, 5, or 6 from onset of natural oestrus, determined by daily behavioural observation and vaginal cytology. Semen was collected from 8 different males by urethral catheterization and electro-ejaculation, during full anaesthesia. Females were immediately immobilized for AI after semen collection. After transrectal ultrasound examination of the reproductive tract, insemination was performed either intravaginal or transcervical using a commercial dog urinary catheter (2.0 × 500 mm, Buster