GxG epistasis in growth and condition and the maintenance of genetic polymorphism in Gambusia holbrooki.

Theory on indirect genetic effects (IGEs) indicates that variation in the genetic composition of social groups can generate GxG epistasis that may promote the evolution of stable polymorphisms. Using a livebearing fish with a genetic polymorphism in coloration and associated behavioral differences, we tested whether genotypes of social partners interacted with focal individual genotypes to influence growth and condition over 16 weeks of development. We found that IGEs had a significant influence on patterns of feeding, regardless of focal fish genotype. There was no influence of social environment on juvenile length, but there was significant GxG epistasis for body condition. Each focal juvenile was in better condition when its own genotype was not present in adult social partners. These data are consistent with negative frequency-dependent selection in which each morph performs better when it is rare. Neither variation in feeding nor activity-related behaviors explained variation in body condition, suggesting that GxG epistasis for condition was caused by physiological differences between the two genotypes. These findings indicate that GxG epistasis in a given polymorphism can generate fitness landscapes that contribute to the maintenance of that polymorphism and to maintenance of genetic variation for additional fitness-related traits.

Contamination rates and antimicrobial resistance in Enterococcus spp., Escherichia coli, and Salmonella isolated from "no antibiotics added"-labeled chicken products.

In the United States, products from chickens that were not administered antimicrobial medications during growout can contain labels stating "no antibiotics added." Here we compared microbial profiles of chicken products labeled as coming from birds raised without antimicrobial medications (N=201; NON) with chicken products carrying conventional labels (N=201; CONV). There were no differences in percentages of samples positive for Enterococcus spp. (CONV: 17.4%; NON: 21.3%) or Escherichia coli (CONV: 25.9%; NON: 22.3%). The number of samples positive for Salmonella was low in both groups, but statistically higher in the NON samples (5.0%) versus CONV samples (1.5%; p<0.05). Conversely, CONV samples contained higher concentrations of coliforms (CONV: 3.0 log(10)CFU/mL; NON: 2.5 log(10)CFU/mL; p<0.05). E. coli (N=190) and Enterococcus spp. isolates (N=113) were tested for resistance to common antimicrobials. E. coli isolates from CONV samples were more frequently resistant to at least one antimicrobial (CONV: 61.3%; NON: 41.2%; p<0.05). Enterococcus spp. isolates from both groups were equally likely to be resistant to at least one antimicrobial, but Enterococcus spp. isolates from CONV samples were more likely to be resistant to erythromycin, kanamycin, and gentamicin (p<0.05). Taken together, these data suggest that NON samples may more frequently carry Salmonella; however, E. coli and Enterococcus spp. found on CONV are more likely to be resistant to some antimicrobials.

Interaction between troponin and myosin enhances contractile activity of myosin in cardiac muscle.

Ca(2+) signaling in striated muscle cells is critically dependent upon thin filament proteins tropomyosin (Tm) and troponin (Tn) to regulate mechanical output. Using in vitro measurements of contractility, we demonstrate that even in the absence of actin and Tm, human cardiac Tn (cTn) enhances heavy meromyosin MgATPase activity by up to 2.5-fold in solution. In addition, cTn without Tm significantly increases, or superactivates sliding speed of filamentous actin (F-actin) in skeletal motility assays by at least 12%, depending upon [cTn]. cTn alone enhances skeletal heavy meromyosin's MgATPase in a concentration-dependent manner and with sub-micromolar affinity. cTn-mediated increases in myosin ATPase may be the cause of superactivation of maximum Ca(2+)-activated regulated thin filament sliding speed in motility assays relative to unregulated skeletal F-actin. To specifically relate this classical superactivation to cardiac muscle, we demonstrate the same response using motility assays where only cardiac proteins were used, where regulated cardiac thin filament sliding speeds with cardiac myosin are >50% faster than unregulated cardiac F-actin. We additionally demonstrate that the COOH-terminal mobile domain of cTnI is not required for this interaction or functional enhancement of myosin activity. Our results provide strong evidence that the interaction between cTn and myosin is responsible for enhancement of cross-bridge kinetics when myosin binds in the vicinity of Tn on thin filaments. These data imply a novel and functionally significant molecular interaction that may provide new insights into Ca(2+) activation in cardiac muscle cells.

Development of an anti-Salmonella phage cocktail with increased host range.

Salmonella shedding in many livestock species can increase significantly after transport and lairage. Preprocessing increases in shedding can amplify the amount of Salmonella that enters the processing facility and the likelihood of end-product contamination. We previously produced an anti-Salmonella phage cocktail that reduced colonization in swine when the pigs were exposed to an environment heavily contaminated with Salmonella, similar to what might be seen in a transport trailer or processing facility holding pen. The aim of this study was to increase the efficacy of the phage treatment by (1) expanding the host-range of the cocktail and (2) developing a more cost-effective microencapsulation technique. We collected samples from wastewater treatment facilities and isolated 20 distinct phages belonging to either the Siphoviridae or Myoviridae families. From this library we identified 10 phages that together lysed a mixed culture of Salmonella enterica Typhimurium, Enteriditis, and Kentucky--three serovars commonly associated with meat and poultry products. The phages were microencapsulated using two sodium-alginate-based methods that only reduced the cocktail titer by 1.0-1.5 logs (premicroencapsulation: 10.4 log(10) PFU/mL; postmicroencapsulation method one: 9.2 log(10) PFU/mL; postmicroencapsulation method two: 8.9 log(10) PFU/mL). Microencapsulated phages remained stable at both 4°C and 22°C for up to 14 days with no appreciable drop in titer (mean titer: 8.9 log(10) PFU/mL). These data indicate that phage cocktails with wider host ranges are possible and a cost-effective microencapsulation process can protect the phages over an extended period, making simultaneous treatment of large numbers of animals with feed- or water-based delivery possible.