The aim of the research was to evaluate two chemical tests for non-invasive pregnancy diagnosis from urine, the Cuboni reaction and the barium chloride test, in donkeys (Equus asinus) and alpacas (Vicugna pacos). The research was carried out from April 2013 to September 2014. Urine samples were collected on five private Czech farms from 18 jennies and 12 alpaca females. Urine was collected non-invasively into plastic cups fastened on a telescopic rod, at 6-9 week intervals. In total, 60 and 54 urine samples from alpacas and jennies, respectively, were collected. The Cuboni reaction was performed by the State Veterinary Institute Prague. The barium chloride test was done with 5 ml of urine mixed together with 5 ml of 1% barium chloride solution. Results of the Cuboni reaction were strongly influenced by the reproductive status of jennies; the test was 100% successful throughout the second half of pregnancy. However, no relationship was found between the real reproductive status of alpaca females and results of the Cuboni reaction. It was concluded that the barium chloride test is not suitable for pregnancy diagnosis either in donkeys, due to significant influence of season on the results, or in alpacas, because no relationship between results of the test and the reproductive status of alpaca females was found. In conclusion, the Cuboni reaction has potential to become a standard pregnancy diagnostic method in donkeys.

Breeding management of small populations may have a critical influence on the development of population characteristics in terms of levels of genetic diversity and inbreeding. Two populations of antelope sister species – Critically Endangered Western Derby eland (Tauroragus derbianus derbianus) and non-native Least Concern Cape eland (Taurotragus oryx oryx) bred under different management strategies were studied in Senegal, Western Africa. The aims of the study were to compare the population genetic parameters of the two species and to test for the presence of interspecific hybrids. In total, blood and tissue samples from 76 Western Derby elands and 26 Cape elands were investigated, using 12 microsatellite markers. No hybrid individuals were detected in the sampled animals within the multispecies enclosure in Bandia Reserve, Senegal. The parameters of genetic polymorphism indicated much lower genetic diversity in Western Derby elands compared to Cape elands. On the other hand, the coefficient of inbreeding was low in both species. It is hypothesized that this could be a positive effect of strict population management of Western Derby elands, which, despite the loss of genetic diversity, minimizes inbreeding.

Experimental non-invasive pregnancy diagnostic techniques, such as seed germination testing in diluted urine, may serve as an alternative to standardized pregnancy diagnostic methods, but scientific validation of these methods is required. This study aimed to use Czech Fleckvieh heifers to investigate the influence of urinary reproductive hormones (oestrone sulphate, 17β-oestradiol, and pregnanediol3-glucuronide) on the germination success of seeds placed within their diluted urine, and further to verify the reliability of seed germination test for pregnancy diagnosis in this species. Mung bean and wheat seeds were germinated in two urine-water dilutions (1:4 and 1:14) for three days, using urine samples obtained from either pregnant or non-pregnant heifers. Germinated seeds were counted daily, and the shoot lengths were measured three days after placement in the urine. Levels of urinary reproductive hormones were determined using competitive heterogeneous enzyme immunoassays (EIAs). Despite the dilution rate used, final results indicated that the urine from pregnant heifers inhibited germination and growth in mung beans only. On the contrary, wheat germination rate was higher within the urine from pregnant females on days 1 and 2, when using a 1:14 dilution rate. For both seed species, correlations between urinary hormone profiles and seed germination parameters were found, depending on the day of the experiment and the rate of dilution used. The shoot lengths of mung bean seeds were significantly (P < 0.05) and negatively correlated with the profiles of all hormones analysed from the urine samples. Thus, urinary reproductive hormones influence seed germination and growth rate of wheat and mung beans, indicating the potential for this technique to be used for pregnancy determination in heifers.

Saliva crystallization was previously studied in both humans and animals with various results. The study aimed to confirm of the presence of saliva crystallization in female Bornean orangutans (Pongo pygmaeus), to evaluate the quality of samples which were collected from animals and processed by keepers, and to test preliminarily if the saliva crystallization could be connected with menstrual cycle and could serve as a cheap, quick and simple method for the basic monitoring of their reproductive status. The research was carried out from September 2014 to January 2015. Sampling of saliva was done in three female orangutans from three zoological gardens (Dvur Kralove, Usti nad Labem, Bojnice) daily, mostly by tongue prints on glass slides with ground edges or by sampling directly from the mouth using plastic spoons from which the saliva was transferred onto glass slides. Samples were evaluated by light microscopy with ×400 magnification. The quality of the sample and type of crystallization was assessed for two different approaches. In total, 246 samples were evaluated. We confirmed the presence of saliva crystallization in orangutans. The quality of samples was variable however acceptable. Unfortunately, it was impossible to detect exact fertile period in two females. However in one orangutan female, when the crystallization was evaluated by the approach typically used in humans, we discovered that saliva crystallization during the fertile period significantly differed from saliva crystallization in the non-fertile period. This points out the possibility of using saliva crystallization for detection of the fertile period in orangutans. However, further research was recommended.

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