Fate and Survival of Campylobacter coli in Swine Manure at Various Temperatures

Campylobacter coli is the most common Campylobacter species found in pig (95%), but the ability of this bacterium to survive in swine manure as well as the potential for causing human illness are poorly understood. We present here laboratory-scale experiments to investigate the effect of temperature on the survival of C. coli in spiked swine manure samples at temperatures from 4 to 52°C. The survival of C. coli during storage for 30 days was studied by three different methods: bacterial culture (plate counting), DNA qPCR, and mRNA RT-qPCR. The results indicate that C. coli could survive in swine manure up to 24 days at 4°C. At higher temperatures, this bacterium survived only 7 days (15°C) or 6 days (22°C) of storage. The survival of C. coli was extremely short (few hours) in samples incubated at 42 and 52°C. The results from the RT-qPCR method were consistent with the data from the bacterial culture method, indicating that it detected only viable C. coli cells, thus eliminating false-positive resulting from DNA from dead C. coli cells

Effect of environmental stress factors on the uptake and survival of Campylobacter jejuni in Acanthamoeba castellanii

<p>Abstract</p> <p>Background</p> <p><it>Campylobacter jejuni</it> is a major cause of bacterial food-borne illness in Europe and North America. The mechanisms allowing survival in the environment and transmission to new hosts are not well understood. Environmental free-living protozoa may facilitate both processes. Pre-exposure to heat, starvation, oxidative or osmotic stresses encountered in the environment may affect the subsequent interaction of <it>C. jejuni</it> with free-living protozoa. To test this hypothesis, we examined the impact of environmental stress on expression of virulence-associated genes (<it>ciaB, dnaJ,</it> and <it>htrA</it>) of <it>C. jejuni</it> and on its uptake by and intracellular survival within <it>Acanthamoeba castellanii</it>.</p> <p>Results</p> <p>Heat, starvation and osmotic stress reduced the survival of <it>C. jejuni</it> significantly, whereas oxidative stress had no effect. Quantitative RT-PCR experiments showed that the transcription of virulence genes was slightly up-regulated under heat and oxidative stresses but down-regulated under starvation and osmotic stresses, the <it>htrA</it> gene showing the largest down-regulation in response to osmotic stress. Pre-exposure of bacteria to low nutrient or osmotic stress reduced bacterial uptake by amoeba, but no effect of heat or oxidative stress was observed. Finally, <it>C. jejuni</it> rapidly lost viability within amoeba cells and pre-exposure to oxidative stress had no significant effect on intracellular survival. However, the numbers of intracellular bacteria recovered 5 h post-gentamicin treatment were lower with starved, heat treated or osmotically stressed bacteria than with control bacteria. Also, while ~1.5 × 10³ colony forming unit/ml internalized bacteria could typically be recovered 24 h post-gentamicin treatment with control bacteria, no starved, heat treated or osmotically stressed bacteria could be recovered at this time point. Overall, pre-exposure of <it>C. jejuni</it> to environmental stresses did not promote intracellular survival in <it>A. castellanii</it>.</p> <p>Conclusions</p> <p>Together, these findings suggest that the stress response in <it>C. jejuni</it> and its interaction with <it>A. castellanii</it> are complex and multifactorial, but that pre-exposure to various stresses does not prime <it>C. jejuni</it> for survival within <it>A. castellanii</it>.</p

From Lab on a Chip to Point of Care Devices: The Role of Open Source Microcontrollers

Microcontrollers are programmable, integrated circuit chips. In the last two decades, their applications to industrial instruments, vehicles, and household appliances have reached the extent that microcontrollers are now the number-one selling electronic chip of all kinds. Simultaneously, the field of lab-on-a-chip research and technology has seen major technological leaps towards sample handling, sample preparation, and sensing for use in molecular diagnostic devices. Yet, the transformation from a laboratory based lab-on-a-chip technology to actual point-of-care device products has largely been limited to a fraction of the foreseen potential. We believe that increased knowledge of the vast possibilities that becomes available with open source microcontrollers, especially when embedded in easy-to-use development environments, such as the Arduino or Raspberry Pi, could potentially solve and even bridge the gap between lab-on-a-chip technology and real-life point of care applications. The profuse availability and extraordinary capabilities of microcontrollers, namely within computation, communication, and networking, combined with easy-to-use development environments, as well as a very active and fast moving community of makers, who are eager to share their knowledge, could potentially be the difference between a dreadful &ldquo;chip-in-a-lab&rdquo;-situation, and the next successful start-up. Here follows a brief insight into how open source microcontrollers could potentially have a transformative effect on the field of lab-on-a-chip research and technology. Details in some specific areas of application are briefly treated before addressing challenges and future perspectives