Effects of temperature on the survival of Tritrichomonas foetus in transport, Diamond's and InPouch TF media

Abstract

The abilities of two isolates of Tritrichomonas foetus to survive and replicate in transport and Diamond's medium or in the InPouch TF system (Bio-Med Diagnostics) when exposed to different temperatures for different periods were determined in a series of experiments. Tubes containing thioglycollate transport medium or pouches were inoculated with 4000 to 5000 organisms and kept for up to seven days at 37 degrees C, 22 degrees C, 4 degrees C, or -20 degrees C. When the holding time had elapsed, the numbers of motile T foetus were counted. Samples in transport medium were transferred to Diamond's medium, and both the pouches and tubes containing Diamond's medium were incubated at 37 degrees C. The cultures were examined and counted four or five times during the 10 to 14 day culture period. The sensitivity of the test under the different conditions, expressed as the number of positive cultures/the total number of samples x 100, varied from zero to 100 per cent depending upon the combination of variables considered. In each medium, with both isolates of T foetus, all samples kept for up to four days at 22 degrees C or 37 degrees C were positive. All cultures of samples kept more than five days at 4 degrees C were negative. No positive cultures were detected when samples were kept more than three hours at -20 degrees C. The day on which the cultures reached mean peak concentration varied with the temperature at which the samples had been kept before they were cultured.

A Review of Some Protozoan Parasites Causing Infertility in Farm Animals

“…Other diagnostic procedures include culture of the organism, polymerase chain reaction (PCR), and immunological tests [58][59][60].…”

Application of a PCR Assay to Enhance the Detection and Identification of Tritrichomonas Foetus in Cultured Preputial Samples

“…Multiple samples were collected from 8 of these bulls for use in this study. Preputial samples were collected by scraping the prepuce with an artificial insemination pipette while applying suction and were inoculated into a thioglycollate transport medium 4 and transported to the laboratory within 1 hour. The samples were centrifuged at 850 ϫ g for 10 minutes and the supernatant removed to leave approximately 0.5 ml or twice the pellet size, whichever was greater.…”

“…The T. foetus organisms (field isolate CAPTF 122) were propagated in modified Diamond's medium 4 to provide flagellates for the creation of spiked samples. Samples for extraction from culture medium were prepared by removing 0.1 ml from serial dilutions of a stock solution of T. foetus organisms in culture.…”

The influence of temperature and simulated transport conditions of diagnostic samples on real-time polymerase chain reaction for the detection ofTritrichomonas foetusDNA

“…Similar culture results were reported in a previous study. 2 The same study reported that cultures samples kept more than 5 days at 4°C gave negative culture results, stressing the assertion that practitioners and diagnosticians must reduce the period in which samples are maintained at 4°C to less than 3 days. Another study found that pre-storage of samples for up to 5 days at 4°C before qPCR would reduce the sensitivity up to 31%.…”

“…5,10 However, accurate PCR results correlate with the quality of the sample, which can be affected by transport condition parameters such as temperature and time of transport to the laboratory. The effects of cold temperatures during transport on the survival of T. foetus using different transport media has been investigated previously by light microscope examination 2 and PCR 4 ; however, previous reports did not consider some of the more extreme high temperature conditions present in tropical and subtropical areas of the country or evaluate the effect on PCR.…”