Prolonged Survival ofCampylobacterSpecies in Bovine Manure Compost

Abstract

The persistence of naturally occurring campylobacteria in aerobic compost constructed of manure from beef cattle that were administered chlortetracycline and sulfamethazine (AS700) or from cattle not administered antibiotics (control) was examined. Although there were no differences in population sizes of heterotrophic bacteria, the temperature of AS700 compost was more variable and did not become as high as that of control compost. There were significant differences in water content, total carbon (C), total nitrogen (N), and electrical conductivity but not in the C/N ratio or pH between the two compost treatments. Campylobacteria were readily isolated from pen manure, for up to day 15 from control compost, and throughout the active phase of AS700 compost. Campylobacter DNA (including Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis, and Campylobacter jejuni) was detected over the ca. 10-month composting period, and no reductions in quantities of C. jejuni DNA were observed over the duration of the active phase. The utilization of centrifugation in combination with ethidium monoazide (EMA) significantly reduced (>90%) the amplification of C. jejuni DNA that did not originate from cells with intact cell membranes. No differences were observed in the frequency of Campylobacter DNA detection between EMA-and non-EMA-treated samples, suggesting that Campylobacter DNA amplified from compost was extracted from cells with intact cell membranes (i.e., from viable cells). The findings of this study indicate that campylobacteria excreted in cattle feces persist for long periods in compost and call into question the common belief that these bacteria do not persist in manure.

Effects of plant species richness and evenness on soil microbial community diversity and function

“…The soil was first incubated using ethidium monoazide bromide (EMA) to prevent the amplification of DNA from nonviable cells (Pisz et al 2007). EMA has been used in a variety of studies to differentiate between viable and non-viable organisms in a variety of environments (Cenciarini-Borde et al 2009;Delgado-Viscogliosi et al 2009;Inglis et al 2010). EMA can intercalate double-stranded DNA and prevent its replication during PCR.…”

The Family Campylobacteraceae

“…Campylobacter spp. are capable of surviving in bovine manure for a 10-month period (Inglis et al 2010). …”

Progress in understanding preferential detection of live cells using viability dyes in combination with DNA amplification

“…The approach is made possible by EMA's efficiency to penetrate dead cells even at relatively low dye concentrations. Whereas early studies employing EMA typically used final dye concentrations of 100 μg/ml (238 μM) (Douglas Inglis et al, 2010;Flekna et al, 2007;Graiver et al, 2010;Marouani-Gradi et al, 2010;Rudi et al, 2005a), concentrations in more recent studies were chosen substantially lower. Cawthorn and Witthuhn (2008) have found that the use of three different EMA concentrations with viable Enterobacter sakazakii cells − 100 (238 μM), 50 (119 μM), and 10 μg/ml (23.8 μM) -resulted in very different PCR product yields of 74, 82 and 92%, respectively (compared with 95, 96, and 97% after PMA treatment).…”