This study is part of a European project focused on understanding the biotic and abiotic mechanisms involved in the retention and dissemination of transmissible spongiform encephalopathies (TSE) infectivity in soil in order to propose practical recommendations to limit environmental contamination. A 1-year field experiment was conducted with lamb carcasses buried in a pasture soil at three depths (25, 45, and 105 cm). Microbial community response to carcasses was monitored through the potential proteolytic activity and substrate induced respiration (SIR). Soil above carcasses and control soil exhibited low proteolytic capacity, whatever the depth of burial. Contrastingly, in soil beneath the carcasses, proteolysis was stimulated. Decomposing carcasses also stimulated SIR, i.e., microbial biomass, suggesting that proteolytic populations specifically developed on lixiviates from animal tissues. Decomposition of soft tissues occurred within 2 months at subsurface while it lasted at least 1 year at deeper depth where proteolytic activities were season-dependent. The ability of soil proteases to degrade the beta form of prion protein was shown in vitro and conditions of burial relevant to minimize the risk of prion protein dissemination are discussed.
cDairy cows have been identified as common carriers of Campylobacter jejuni, which causes many of the human gastroenteritis cases reported worldwide. To design on-farm management practices that control the human infection sourced from dairy cows, the first step is to acquire an understanding of the excretion patterns of the cow reservoir. We monitored the same 35 cows from two dairy farms for C. jejuni excretion fortnightly for up to 12 months. The objective was to examine the concentration of C. jejuni and assess the genetic relationship of the C. jejuni populations excreted by individual cows. Significant differences (P < 0.01) in C. jejuni fecal concentration were observed among the 35 cows, with median concentrations that varied by up to 3.6 log 10 · g ؊1 feces. A total of 36 different genotypes were identified from the 514 positive samples by using enterobacterial repetitive intergenic consensus (ERIC)-PCR. Although 22 of these genotypes were excreted by more than one cow, the analysis of frequencies and distribution of the genotypes by model-based statistics revealed a high degree of individuality in the C. jejuni population in each cow. The observed variation in the frequency of excretion of a genotype among cows and the analysis by multilocus sequence typing (MLST) of these genotypes suggest that excretion of C. jejuni in high numbers is due to a successful adaptation of a particular genotype to a particular cow's gut environment, but that animal-related factors render some individual cows resistant to colonization by particular genotypes. The reasons for differences in C. jejuni colonization of animals warrant further investigation.
Summary There is an increasing interest in the detection and enumeration of micro‐organisms pathogenic for human and present in bovine faeces. This interest is because pollution of the environment by animal faeces may affect the safety of food and of drinking or recreational water. Detection and quantification of microbial pathogens carried out using DNA extracted from the faecal matrix are affected by the quality and the quantity of the DNA extracts, which are critical factors that limit the accuracy and sensitivity of molecular studies. This review compares published methods on DNA extraction from bovine faeces, focusing on the extent to which the success of DNA amplification is affected by issues related to the faeces. Following a general discussion on the DNA extraction methods used for faeces, we focus particularly on issues related to the faecal environment itself. The objective is to identify information that can be used to improve the sensitivity of those PCR methods used after direct DNA extraction.
The objective of this study was to compare the health, physiology, and behavior of group-housed calves reared on wood shavings with those reared on alternative surfaces. At 1 wk of age, 80 calves were moved into 1 of 20 experimental pens (n = 4 calves/pen) where they remained until 6 wk of age. Pens had floors covered with pea gravel (PG), rubber chip (RC), sand (SA), or wood shavings (WS; n = 5 pens/substrate). Body weight, cleanliness, health, and skin surface and vaginal temperature were recorded at 1, 3, and 6 wk of age. Escherichia coli numbers were assessed on the skin surface of the shoulder and in the feces of calves at 3 and 6 wk of age. Blood samples were taken at 1, 3, and 6 wk of age to measure hematological values and cortisol, IgG, and lactate concentrations. Behaviors (lying, running, and self-grooming) were recorded in the home pen at 1, 3, and 6 wk of age using video recorders and accelerometer data loggers. At 6 wk of age, calves were tested individually in an arena test and behavior was recorded continuously for 20 min. Body weight did not differ among calves reared on PG, RC, SA, or WS, regardless of age. All calves were clean and no calves displayed any signs of lameness, leg lesions, or injuries at wk 1, 3, or 6, regardless of substrate. The number of E. coli recovered from a surface area of 100 cm on the shoulder of each calf was affected by rearing substrate, with more E. coli recovered from calves reared on WS than PG, RC, or SA at 3 and 6 wk of age. Fecal E. coli counts were not affected by rearing substrate at 3 or 6 wk of age. Over the entire study period, calves reared on PG and SA had lower skin temperatures than calves reared on RC or WS, but skin temperature was similar between calves reared on PG and SA. However, vaginal temperature did not differ among calves reared on different substrates at 1, 3, or 6 wk of age. Hematology values and cortisol, IgG, and lactate concentrations of calves were similar among rearing substrates over the 6-wk study period. In the home pen, rearing substrate did not influence time spent lying; however, calves reared on WS performed more lying bouts than calves reared on PG or SA. In addition, rearing substrate did not influence the time calves spent running; however, calves reared on WS spent more time self-grooming than calves reared on PG, RC, and SA. During a 20-min arena test, running, bucks, jumps, and kicks performed by calves was not affected by rearing substrate. In conclusion, the physiology and behavior of calves reared on PG, RC, and SA was similar to WS, which is considered the preferred rearing substrate to use when rearing calves. Therefore, PG, RC, and SA may be acceptable substrate options when rearing group-housed dairy calves.
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