ABSTRACT:Mannheimia haemolytica consistently causes severe bronchopneumonia and rapid death of bighorn sheep (Ovis canadensis) under experimental conditions. However, Bibersteinia trehalosi and Pasteurella multocida have been isolated from pneumonic bighorn lung tissues more frequently than M. haemolytica by culture-based methods. We hypothesized that assays more sensitive than culture would detect M. haemolytica in pneumonic lung tissues more accurately. Therefore, our first objective was to develop a PCR assay specific for M. haemolytica and use it to determine if this organism was present in the pneumonic lungs of bighorns during the 2009-2010 outbreaks in Montana, Nevada, and Washington, USA. Mannheimia haemolytica was detected by the species-specific PCR assay in 77% of archived pneumonic lung tissues that were negative by culture. Leukotoxin-negative M. haemolytica does not cause fatal pneumonia in bighorns. Therefore, our second objective was to determine if the leukotoxin gene was also present in the lung tissues as a means of determining the leukotoxicity of M. haemolytica that were present in the lungs. The leukotoxin-specific PCR assay detected leukotoxin gene in 91% of lung tissues that were negative for M. haemolytica by culture. Mycoplasma ovipneumoniae, an organism associated with bighorn pneumonia, was detected in 65% of pneumonic bighorn lung tissues by PCR or culture. A PCR assessment of distribution of these pathogens in the nasopharynx of healthy bighorns from populations that did not experience an all-age die-off in the past 20 yr revealed that M. ovipneumoniae was present in 31% of the animals whereas leukotoxin-positive M. haemolytica was present in only 4%. Taken together, these results indicate that culture-based methods are not reliable for detection of M. haemolytica and that leukotoxin-positive M. haemolytica was a predominant etiologic agent of the pneumonia outbreaks of 2009-2010.
Studies addressing prehistoric mobility in animals typically use isotopic analyses of sequentially formed tissues, such as the growth layers in teeth, to infer physical movement on the landscape. Strontium isotope ratios (87Sr/86Sr values), which vary geographically, are particularly useful for this purpose, especially when paired with stable oxygen isotope ratios (δ18O), which vary seasonally. Together, these two isotope systems can provide information about past animal movement patterns on a seasonal scale. However, while many studies have used 87Sr/86Sr and δ18O values from analyses of sequentially formed tissues for this purpose, there have been limited analyses on modern animals of known movement patterns across high-latitude regions. In this pilot study, we sequentially sampled and analyzed one second molar (M2) and two third molars (M3) from two bison (Bison bison bison) from the Delta bison herd, which resides in interior Alaska and has known and documented seasonal mobility patterns. The resulting 87Sr/86Sr values from the teeth were compared to a high-resolution 87Sr/86Sr isoscape for the region and were paired with δ18O analyses to determine whether the seasonal 87Sr/86Sr values matched the predicted values for each of the seasonal bison habitat areas. The results indicate that the 87Sr/86Sr and δ18O values reliably reflected the known seasonal mobility patterns of bison and suggest that this approach could be used to investigate the mobility patterns of prehistoric bison in Alaska and surrounding high-latitude regions.
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