Skip to main content Accessibility help

Addressing the constraints of Tritrichomonas foetus sample collection in remote areas: lyophilized modified Diamond's media as a substitute for liquid medium

  • Gemma Rush (a1), Michael William Reynolds (a1), Nichola Eliza Davies Calvani (a1) and Jan Šlapeta (a1)


Bovine trichomoniasis is a notifiable, reproductive disease of cattle caused by the parasite Tritrichomonas foetus. Culturing with modified Diamond's medium (MDM) is required to increase the low number of organisms received from a preputial sample, but is limited in application to remote areas as it requires continuous cold chain storage. This study utilized lyophilization to sustain the viability of MDM during transport in lieu of a continuous cold chain. All lyophilized MDM was able to sustain T. foetus after storage for 42 days at 24 °C, and the results demonstrated that lyophilized MDM was equally as viable as refrigerated liquid MDM. Storage of lyophilized MDM at room temperature for 1 and 7 days did not impact T. foetus yield, both with and without exposure to light. A limitation of the lyophilized MDM was demonstrated with a significant decrease in T. foetus yield when the media was stored at 37 and 58 °C. The lyophilization of MDM provides a robust method of transporting and storing medium prior to reconstitution and inoculation, for use in T. foetus diagnosis and surveillance in remote areas.


Corresponding author

Author for correspondence: Jan Šlapeta, E-mail:


Hide All
Balczun, C and Scheid, PL (2018) Lyophilisation as a simple and safe method for long-term storage of free-living amoebae at ambient temperature. Parasitology Research 117, 33333336.
Bernasconi, Ch, Bodmer, M, Doherr, MG, Janett, F, Thomann, A, Spycher, C, Iten, C, Hentrich, B, Gottstein, B, Müller, N and Frey, CF (2014) Tritrichomonas foetus: prevalence study in naturally mating bulls in Switzerland. Veterinary Parasitology 200, 289294.
Biomed Diagnostics (n.d.) InPouchTF (Tritrichomonas foetus Bovine). Retrieved from Biomed Diagnostics: (accessed 28 August 2018).
Bjelošević, M, Selja, KB, Trstenjak, U, Logar, M, Brus, B and Grabnar, PA (2018) Aggressive conditions during primary drying as a contemporary approach to optimise freeze-drying cycles of biopharmaceuticals. European Journal of Pharmaceutical Sciences 122, 292302.
Buller, N and Corney, B (2013) Bovine Trichomoniasis: Australian and New Zealand Standard Diagnostic Procedure. Canberra, Australia: Australian Government, Department of Agriculture and Water Resources.
Cobo, ER, Favetto, PH, Lane, VM, Friend, A, VanHooser, K, Mitchell, J and BonDurant, RH (2007) Sensitivity and specificity of culture and PCR of smegma samples of bulls experimentally infected with Tritrichomonas foetus. Theriogenology 68, 853860.
Collantes-Fernandez, E, Mendoza-Ibarra, JA, Pedraza-Diaz, S, Rojo-Montejo, S, Navarro-Lozano, V, Sanchez-Sanchez, R, Ruiz-Santa-Quiteria, JA, Ortega-Mora, LM and Osoro, K (2014) Efficacy of a control program for bovine trichomonosis based on testing and culling infected bulls in beef cattle managed under mountain pastoral systems of Northern Spain. The Veterinary Journal 200, 140145.
Duralliu, A, Matejtschuk, P and Williams, DR (2018) Humidity induced collapse in freeze-dried cakes: a direct visualisation study using DVS. European Journal of Pharmaceutics and Biopharmaceutics 127, 2936.
Eccles, J (2016) Trichomoniasis in the Northern Territory: AgNote. Retrieved from Northern Territory Government: (accessed 10 September 2018).
Gookin, JL, Hanrahan, K and Levy, MG (2017) The conundrum of feline trichomonosis – the more we learn the ‘trickier’ it gets. Journal of Feline Medicine and Surgery 19, 261274.
Holzer, M, Vogel, V, Mantele, W, Schwartz, D, Haase, W and Langer, K (2009) Physico-chemical characterisation of PLGA nanoparticles after freeze-drying and storage. European Journal of Pharmaceutics and Biopharmaceutics 72, 428437.
Jensen, JB (1983) In Vitro Cultivation of Protozoan Parasites. Boca Raton, FL: CRC Press.
Levine, ND (1973) Protozoan Parasites of Domestic Animals and of Man, 2nd Edn. Minneapolis, MN: Burgess Publishing Company.
Lun, ZR, Parker, S and Gajadhar, AA (2000) Comparison of growth rates of Tritrichomonas foetus isolates from various geographic regions using three different culture media. Veterinary Parasitology 89, 199208.
Marco-Jimenez, F, Garzon, DL, Penaranda, DS, Perez, L, Viudes-de-Castro, MP, Vicente, JS, Jover, M and Asturiano, JF (2006) Cryopreservation of European eel (Anguilla anguilla) spermatozoa: effect of dilution ratio, foetal bovine serum supplementation, and cryoprotectants. Cryobiology 53, 5157.
McCool, CJ, Townsend, MP, Wolfe, SG, Simpson, MA, Olm, TC, Jayawardhana, GA and Carney, JV (1988) Prevalence of bovine venereal disease in the Victoria River District of the Northern Territory: likely economic effects and practicable control measures. Australian Veterinary Journal 65, 153156.
Mueller, K, Morin-Adeline, V, Gilchrist, K, Brown, G and Šlapeta, J (2015) High prevalence of Tritrichomonas foetus ‘bovine genotype’ in faecal samples from domestic pigs at a farm where bovine trichomonosis has not been reported for over 30 years. Veterinary Parasitology 212, 105110.
Mukhufhi, N, Irons, PC, Michel, A and Peta, F (2003) Evaluation of a PCR test for the diagnosis of Tritrichomonas foetus infection in bulls: effects of sample collection method, storage and transport medium on the test. Theriogenology 60, 12691278.
National Farmer's Federation (2017) Food, Fibre and Forestry Facts: A Summary of Australia's Agricultural Sector. Retrieved from National Farmer's Federation, Australia: (accessed 2 September 2018).
Nims, RW and Harbell, JW (2017) Best practices for the use and evaluation of animal serum as a component of cell culture medium. In Vitro Cellular & Developmental Biology – Animal 53, 682690.
OIE (2018) OIE-Listed diseases, infections and infestations in force in 2018. Retrieved from OIE: (accessed 20 August 2018).
OIE Terrestrial Manual (2012) Trichomonosis. Retrieved from OIE Terrestrial Manual 2012, Chapter 2.4.16: (accessed 20 August 2018).
Šlapeta, J, Craig, S, McDonell, D and Emery, D (2010) Tritrichomonas foetus from domestic cats and cattle are genetically distinct. Experimental Parasitology 126, 209213.
Šlapeta, J, Müller, N, Stack, CM, Walker, G, Lew-Tabor, A, Tachezy, J and Frey, CF (2012) Comparative analysis of Tritrichomonas foetus (Riedmüller, 1928) cat genotype, T. foetus (Riedmüller, 1928) cattle genotype and Tritrichomonas suis (Davaine, 1875) at 10 DNA loci. International Journal for Parasitology 42, 11431149.
Tang, Z and Pikal, MJ (2004) Design of freeze-drying processes for pharmaceuticals: practice advice. Pharmaceutical Research 21, 191200.



Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed