Efficacy of an Anaerobic Swab Transport System to Maintain Aerobic and Anaerobic Microorganism Viability after Storage at −80°C

Musser, Jeffrey M.B.; Gonzalez, R. Gilberto

doi:10.1177/104063871102300115

Cited by 11 publications

(7 citation statements)

References 20 publications

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“…We assume that the lack of cooling during the transport to laboratory C might have affected agreement. It is known that temperature, time, and type of swab (Roelofsen et al, 1999), media (Yrios et al, 1975), and transport system (Rishmawi et al, 2007;Stoner et al, 2008;Musser and Gonzalez, 2011) affect viability and stability of microorganisms. Effects of our transport medium on our 4 target pathogens have not been examined yet.…”

Section: Methodsmentioning

confidence: 99%

Relationship between bacteriological findings in the second and fourth weeks postpartum and uterine infection in dairy cows considering bacteriological results

Werner

Suthar

Plöntzke

et al. 2012

Journal of Dairy Science

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The uterine lumen in early postpartum dairy cows is contaminated with different bacteria. The most relevant uterine pathogens are Escherichia coli and Trueperella pyogenes. Prevalence of α-hemolytic streptococci and coagulase-negative staphylococci (CNS) is also high; however, these pathogens are considered opportunistic. The overall objective of this study was to investigate effects of the intrauterine presence of E. coli, T. pyogenes, α-hemolytic streptococci, or CNS at 10±1 d in milk (DIM) on the type of bacteria 2 wk later and their influence on uterine infections and subsequent reproductive performance. Furthermore, we set out to quantify 2 relevant methodological factors (i.e., laboratory and sampling instrument). Bacteriological samples were collected at 10±1 and 24±1 DIM from the uterine lumen using a cytobrush (CB). Vaginal mucus was classified by vaginoscopy. In a subsample, bacteriological results of 3 different laboratories and of CB and cotton swabs (CS) were compared. Samples of uterine discharge were collected at 10±1 DIM and bacteriological samples were taken using CB and CS. Bacteria were identified and bacterial growth quantified on a 4-point scale. Animals infected with E. coli or T. pyogenes at 10±1 DIM had a higher risk for an infection with the same bacterial species at 24±1 DIM [E. coli relative risk (RR)=3.7 and T. pyogenes RR=2.9]. Moreover, the risk of being diagnosed with abnormal vaginal discharge at 24±1 DIM increased in cows with E. coli (RR=1.7) or T. pyogenes (RR=1.7) at 10±1 DIM. Uterine infection with α-hemolytic streptococci or CNS did not increase the risk of an infection with T. pyogenes or E. coli or abnormal vaginal discharge 2 wk later. Cows with E. coli at 10±1 DIM or T. pyogenes at 24±1 DIM had greater days to first artificial insemination than cows positive for the 3 remaining bacterial species. Cows with T. pyogenes at 10±1 DIM had more days to pregnancy and more cows were culled when positive for E. coli at 10±1 DIM. Agreement of bacteriological results of 3 different laboratories were significant for laboratory A + B and A + C for the CB and CS. The highest agreement considering the bacterial species was for E. coli. All results for laboratory A + B and A + C for the CB and CS were significant. The results generated from samples collected with CB agreed nicely with those from CS from each laboratory (laboratory A: 250/272; laboratory B: 264/272; laboratory C: 253/272).

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Section: Methodsmentioning

confidence: 99%

Relationship between bacteriological findings in the second and fourth weeks postpartum and uterine infection in dairy cows considering bacteriological results

Werner

Suthar

Plöntzke

et al. 2012

Journal of Dairy Science

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“…However, this is a logistic challenge in large cohort studies which usually rely on fecal swabs [54]. Fecal swabs are commonly stored for short-term at room temperature in specific media in which obligate anaerobes can survive [55].…”

Section: How To Study and Interpret Microbiotamentioning

confidence: 99%

The gut microbiota of nonalcoholic fatty liver disease: current methods and their interpretation

2015

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The role of intestinal bacteria in the pathogenesis of nonalcoholic fatty liver disease is increasingly acknowledged. Recently developed microbial profiling techniques are beginning to shed light on the nature of gut microbiota alterations in nonalcoholic fatty liver disease. In this review, we summarize the gut microbiota composition changes that have been reported during different stages of human nonalcoholic fatty liver disease, and highlight the relation between bile acids and gut bacteria in this context. In addition, we discuss the different methodologies used in microbiota analyses as well as the interpretation of microbiota data. Whereas the currently available studies have provided useful information, future large-scale prospective studies with carefully phenotyped subjects and sequential sampling will be required to demonstrate a causal role of gut microbiota changes in the etiology of nonalcoholic fatty liver disease.

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“…Along with storage temperature, other user friendly sampling methods should be investigated, which are relatively easy to implement in large cohorts or when including consecutive patients from daily clinical practice. Fecal swabs are routinely used in clinical settings to detect enteropathogens and multidrug resistant Enterobacteriaceae, proving the feasibility of this method [ 10 , 11 ]. Since it is claimed that some of these swabs are also suitable for molecular analysis, it is conceivable that they might also be usable to study the fecal microbiota composition by means of next generation sequencing.…”

Section: Introductionmentioning

confidence: 99%

The Effect of Sampling and Storage on the Fecal Microbiota Composition in Healthy and Diseased Subjects

et al. 2015

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Large-scale cohort studies are currently being designed to investigate the human microbiome in health and disease. Adequate sampling strategies are required to limit bias due to shifts in microbial communities during sampling and storage. Therefore, we examined the impact of different sampling and storage conditions on the stability of fecal microbial communities in healthy and diseased subjects. Fecal samples from 10 healthy controls, 10 irritable bowel syndrome and 8 inflammatory bowel disease patients were collected on site, aliquoted immediately after defecation and stored at -80°C, -20°C for 1 week, at +4°C or room temperature for 24 hours. Fecal transport swabs (FecalSwab, Copan) were collected and stored for 48-72 hours at room temperature. We used pyrosequencing of the 16S gene to investigate the stability of microbial communities. Alpha diversity did not differ between all storage methods and -80°C, except for the fecal swabs. UPGMA clustering and principal coordinate analysis showed significant clustering by test subject (p<0.001) but not by storage method. Bray-Curtis dissimilarity and (un)weighted UniFrac showed a significant higher distance between fecal swabs and -80°C versus the other methods and -80°C samples (p<0.009). The relative abundance of Ruminococcus and Enterobacteriaceae did not differ between the storage methods versus -80°C, but was higher in fecal swabs (p<0.05). Storage up to 24 hours (at +4°C or room temperature) or freezing at -20°C did not significantly alter the fecal microbial community structure compared to direct freezing of samples from healthy subjects and patients with gastrointestinal disorders.

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Efficacy of an Anaerobic Swab Transport System to Maintain Aerobic and Anaerobic Microorganism Viability after Storage at −80°C

Cited by 11 publications

References 20 publications

Relationship between bacteriological findings in the second and fourth weeks postpartum and uterine infection in dairy cows considering bacteriological results

Relationship between bacteriological findings in the second and fourth weeks postpartum and uterine infection in dairy cows considering bacteriological results

The gut microbiota of nonalcoholic fatty liver disease: current methods and their interpretation

The Effect of Sampling and Storage on the Fecal Microbiota Composition in Healthy and Diseased Subjects

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