Evaluation of radiometric faecal culture and direct PCR on pooled faeces for detection of Mycobacterium avium subsp. paratuberculosis in cattle

Dilution rates for pooled faecal culture (PFC) and direct IS900 polymerase chain reaction (D-PCR) tests were evaluated on faecal samples from infected cows mixed with uninfected faeces in dilutions from 1 in 5 to 1 in 50. PFC was performed by radiometric culture, with confirmation by IS900 PCR and restriction endonuclease analysis (PCR/REA) on growth, and by mycobactin dependency testing on solid medium. Using 37 culture positive faecal samples from 12 subclinical cows, 83.8% and 94.6% of samples were confirmed positive in the PFC assay at dilutions of 1 in 50 and 1 in 30, respectively. Lower dilutions (1 in 5 to 1 in 20) provided only marginally better sensitivity, and confirmation of PFC growth by PCR/REA was significantly more sensitive than mycobactin dependency. D-PCR had significantly lower sensitivity than PFC confirmed by PCR/REA, with pools of 1 in 50, 30, 10 and 5 yielding positive results in 64.9%, 70.3%, 78.4% and 83.8% of samples, respectively. Cattle considered to be shedding 1.5 x 10(6) viable M. avium subsp. paratuberculosis (Map)/g faeces (on the basis of estimated losses in processing and growth rates in radiometric broth) were positive at dilutions up to 1 in 50 in the PFC and D-PCR. Those shedding 5 x 10(5) viable Map/g were positive in the PFC at dilutions up to 1 in 40, but required a 1 in 10 dilution or less for D-PCR. The results suggest that for cattle shedding relatively high concentrations of Map in faeces (>2 x 10(5) viable Map/g), maximal dilutions of 1 in 30 for PFC and 1 in 10 for D-PCR would be applicable.     

Sampling and repeatability of radiometric faecal culture in bovine Johne's disease

The repeatability of detection of Mycobacterium avium subsp. paratuberculosis (Map) within and between samplings from 16 paratuberculous dairy cows (13 subclinical; 3 clinical) was investigated by radiometric culture of quadrants of faecal dung pats collected on four to seven occasions over a 10-16-day period. Results were compared to serological status and to pathological and bacteriological findings in multiple tissues obtained at slaughter from 15 of the animals 2-6 weeks after the faecal samplings. From faecal samples taken on 77 occasions over the 2-week period, 296/308 (96%) quadrants were culture positive, with samples from all cattle showing evidence of faecal shedding of Map. Histological lesions typical of paratuberculosis were present in 14 of the 15 cows examined at slaughter, varying in severity from mild (two animals) to moderate (4) and advanced (8), and all predilection tissue sites yielded Map. The negative faecal samples were derived from a single animal that was culture positive in two quadrants on each of the first two (of four) sampling occasions (i.e. culture positive in only 4 of 16 collected quadrants). This animal was found to be histologically negative at slaughter, and culture positive from three of five predilection tissue sites. Faecal samples from cows with subclinical and clinical paratuberculosis, with lesion severity ranging from mild to severe at multiple predilection sites, produced faeces with relatively consistent concentrations of Map within samples. There was significant variation in concentrations of Map between samples in individual animals over a period of 2 weeks, but this did not affect the dichotomous positive-negative culture status for 15 of the 16 cattle. A faecal sample collected non-randomly per rectum thus provides a representative specimen for detection of Map by radiometric culture on a single sampling occasion.     

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Historical influences dominate the composition of regenerating plant communities in abandoned citrus groves in north-central Florida

The question of what determines plant community composition is fundamental to the study of plant community ecology. We examined the relative roles of historical land use, landscape context, and the biophysical environment as determinants of plant community composition in regenerating citrus groves in north-central Florida. Results were interpreted in light of plant functional traits. Herbaceous and woody plants responded differently to broad-scale variables; herbs correlated most strongly with surrounding land cover at a scale of 8 km, while the only significant determinant of woody species distributions was local land use history. There were significant correlations between herbaceous species and spatial context, habitat isolation, environmental variables, and historical variables. Partial Mantel tests indicated that each variable provided a unique contribution in explaining some of the variation in the herbaceous dataset. The correlation between woody plants and local historical variables remained significant even with other effects corrected for. In the herbaceous community, species composition was linked to functional traits much as expected from classical theory. While spatial influences in our study system are important for both woody and herbaceous plants, the primary determinant of plant community composition in regenerating citrus groves is historical land use. Our results suggest that the fine-scale mechanisms of local competition, tolerance and facilitation invoked by many classical studies may ultimately be less important than land use history in understanding current plant community composition in regenerating agricultural areas.     

Reflectance of Alaskan black spruce and white spruce foliage in relation to elevation and latitude

Leaf reflectance at visible and near-infrared wavelengths (400-1000 nm) is related primarily to pigmentation, leaf structure and water content, and is an important tool for studying stress physiology and relationships between plants and their growth environment. We studied reflectance of two co-occurring Alaskan conifers, black spruce (Picea mariana (Mill.) BSP) and white spruce (Picea glauca (Moench) Voss), at elevations from 60 to 930 m a.s.l. along a latitudinal gradient from 61 degrees to 68 degrees N. Black spruce samples were collected from 24 sites and white spruce from 30 sites. Overall, reflectance spectra of the two species were similar, but from 400 to 700 nm, needle reflectance was consistently higher in black spruce than in white spruce (all P 相似文献