A review of antimicrobial peptides: defensins and related cationic peptides

Cationic antimicrobial peptides are present throughout the plant and animal kingdoms and bear striking structural and functional similarities across species lines. They provide primitive, nonspecific means of combating a variety of bacteria, fungi, enveloped viruses, and protozoa. Some are also cytotoxic against host cells, including neoplastic cells. Cationic antimicrobial peptides may play various roles in inflammation and tissue repair. Antimicrobial peptides are found in epithelial tissues regularly exposed to microbial attack as well as in cells whose primary function is defense against potential pathogens. They constitute an important part of the nonoxidative antimicrobial arsenal of leukocytes. They are preformed and/or readily synthesized when the cells are stimulated by exposure to pathogens. They exert their effects directly by inserting into membranes of target cells and forming ion channels which increase membrane permeability; however, antimicrobial peptides can also act as opsonins to facilitate phagocytosis. Resistance to defensins is a virulence factor for organisms such as Salmonella sp. The study of cationic antimicrobial peptides is increasing our understanding of innate immunity, inflammation, and the pathogenesis of genetic diseases such as specific granule disease in humans. Therapeutic applications of antimicrobial peptides are currently under investigation.     

Evaluation of low-ash poultry meal as a protein source in canine foods

Eight mature female dogs (18.0 +/- 0.2 kg) were used in a replicated 4 x 4 Latin square experiment to determine the feeding value of low-ash poultry meal (PM) in a complete food fed to dogs. All foods contained graded concentrations of PM (10.4 to 32.5% DM), resulting in foods that were 10, 15, 20, and 25% CP. Daily DMI averaged 284 +/- 14 g/d. An increase in PM resulted in an increase in fecal moisture from 44.7 to 55.1% (linear; P < 0.01), and fecal DM output increased from 24.8 to 31.6 g/d (linear; P < 0.05). Ileal DM flow increased from 27.1 to 40.7 g/d (linear; P < 0.01). Small intestinal DM digestibility decreased from 90.4 to 86.1% (linear; P < 0.01) and total-tract DM digestibility decreased from 91.2 to 89.4% (linear; P < 0.01) as PM increased. Large intestinal DM digestibility increased from 8.4 to 21.1% with increasing PM (linear; P < 0.05). Fecal excretion of CP increased from 5.6 to 10.0 g/d (linear; P < 0.01) and ileal flow of CP increased from 6.9 to 15.6 g/d (linear; P < 0.01) as PM increased. Small intestinal CP digestibility was unaffected with treatment (P > 0.05). Large intestinal CP digestibility increased from 21.6 to 37.1% (linear; P < 0.05) with increasing PM. Total-tract CP digestibility increased from 81.0 to 86.6% (linear; P < 0.01) as PM increased. Arginine had the highest overall digestibility ranging from 88.5 to 91.3%, whereas cysteine had the lowest digestibility, ranging from 67.1 to 71.4%. These data indicate that PM is a highly digestible protein source for canine foods with inclusions of 10.4 to 32.5% of DM.     

Compositional Shifts in Forb and Butterfly Communities Associated with Kentucky Bluegrass Invasions

Rangeland management strategies impact biodiversity, the quality and quantity of ecosystem services, and overall rangeland resiliency. Previous management strategies, coupled with climate change, have led to widespread invasion by Kentucky bluegrass (Poa pratensis; bluegrass) in the Northern Great Plains, United States. Bluegrass invasions are expected to have detrimental impacts on biodiversity conservation and ecosystem services provided by rangelands. Yet none have investigated how bluegrass invasions influence pollinator populations, which are a prominent conservation concern and provide ecosystem services. We measured the impact of bluegrass invasion on mixed-grass prairie forb and butterfly communities. Obligate grassland butterflies, those that rely on grasslands, decreased as bluegrass cover increased, including the threatened Hesperia dacotae. Conversely, the abundance of facultative grassland butterflies, those found in grasslands but not fully dependent on them for their life history, increased as bluegrass increased. Moreover, plant species diversity and flowering forb species richness decreased as bluegrass cover increased. Overall, bluegrass invasion led to butterfly and plant community simplification, signaling a loss of biodiversity and potentially ecosystem services. Our research is the first to quantify how grassland butterflies and the floral resources they depend on are negatively impacted by bluegrass invasion. Resource managers should adopt management strategies that reduce bluegrass cover and improve nectar and host resources for obligate grassland butterflies. Management choices that removed disturbance regimes inherent to the Northern Great Plains (i.e., fire and grazing) led to bluegrass dominance in the region. Therefore, restoring disturbance regimes may be one way to reduce bluegrass and benefit pollinator populations.     

Influence of slow-release urea on nitrogen balance and portal-drained visceral nutrient flux in beef steers

Two experiments were conducted to evaluate the effects of slow-release urea (SRU) versus feed-grade urea on portal-drained visceral (PDV) nutrient flux, nutrient digestibility, and total N balance in beef steers. Multi-catheterized steers were used to determine effects of intraruminal dosing (Exp. 1; n = 4; 319 +/- 5 kg of BW) or feeding (Exp. 2; n = 10; 4 Holstein steers 236 +/- 43 kg of BW and 6 Angus steers 367 +/- 46 kg of BW) SRU or urea on PDV nutrient flux and blood variables for 10 h after dosing. Intraruminal dosing of SRU (Exp. 1) prevented the rapid increase in ruminal ammonia concentrations that occurred with urea dosing (treatment x time P = 0.001). Although apparent total tract digestibilities of DM, OM, NDF, and ADF were not affected by treatment (P > 0.53, Exp. 2), SRU increased fecal N excretion (49.6 vs. 45.6 g/d; P = 0.04) and reduced apparent total tract N digestibility (61.7 vs. 66.0%; P = 0.003). Transfer of urea from the blood to the gastrointestinal tract occurred for both treatments in Exp. 1 and 2 at all time points with the exception for 0.5 h after dosing of urea in Exp. 1, when urea was actually transferred from the gastrointestinal tract to the blood. In both Exp. 1 and 2, both urea and SRU treatments increased arterial urea concentrations from 0.5 to 6 h after feeding, but arterial urea concentrations were consistently less with SRU (treatment x time P < 0.001, Exp. 1; P = 0.007, Exp. 2). Net portal ammonia release remained relatively consistent across the entire sampling period with SRU treatment, whereas urea treatment increased portal ammonia release in Exp. 1 and tended to have a similar effect in Exp. 2 (treatment x time P = 0.003 and P = 0.11, respectively). Urea treatment also increased hepatic ammonia uptake within 0.5 h (treatment x time P = 0.02, Exp. 1); however, increased total splanchnic release of ammonia for the 2 h after urea treatment dosing suggests that PDV ammonia flux may have exceeded hepatic capacity for removal. Slow-release urea reduces the rapidity of ammonia-N release and may reduce shifts in N metabolism associated with disposal of ammonia. However, SRU increased fecal N excretion and increased urea transfer to the gastrointestinal tract, possibly by reduced SRU hydrolysis or effects on digestion patterns. Despite this, the ability of SRU to protect against the negative effects of urea feeding may be efficacious in some feeding applications.     

Improved methods for isolating DNA from Ostertagia ostertagi eggs in cattle feces

A multiplex PCR assay for differentiating strongyle eggs from cattle has recently been described; however, the egg disruption and DNA extraction procedures, though effective, are inadequate for large studies or clinical application. The purpose of this research was to evaluate methods for disrupting trichostrongyle eggs, then assess commercial kits for extracting egg DNA using Ostertagia ostertagi as a model species. Egg disruption procedures tested included probe sonication, bath sonication, bead beating, boiling, microwaving, proteinase K/SDS digestion, freezing, and various combinations of the above with the incorporation of sodium dodecyl sulfate. These procedures were evaluated in conjunction with four commercial DNA extraction kits: DNA Stool mini kit and DNeasy Plant kit (Qiagen), Fast DNA kit (QBiogene), and the MAP extraction kit (Tetracore). Results showed that egg disruption was best accomplished with the bead beater and ceramic beads, resulting in 100% disruption within 1min. When DNA extraction was preceded by the isolation of eggs from feces, all procedures except the Fast DNA kit produced PCR-ready DNA from at least two eggs. The DNeasy Plant kit allowed consistent detection of DNA released from one egg. Due to the morphological similarities among trichostrongyle eggs in ruminants, strongyle eggs in equids, and hookworm eggs, the methods described herein may have broad application to other nematodes.     

Detecting defects in conifers with ground penetrating radar: applications and challenges

Our objective was to test ground penetrating radar (GPR) to non‐destructively estimate decay volumes in living coniferous trees. GPR is geophysical tool which uses an antenna to propagate short bursts of electromagnetic energy in solid materials and measure the two‐way travel time and amplitude of reflected signals. We compared estimates of bole decay from data collected with a SIR 3000 GPR system equipped with a 900 MHz antenna to measurements of decay from stem cross sections and increment cores for three conifer species (Pseudotsuga menziesii, Thuja plicata and Tsuga heterophylla). We found that near‐surface decay, air‐filled voids and desiccated boles had unique electromagnetic signatures, which could be separated from other defects. GPR successfully estimated the percent area of air‐filled cavities and was not significantly different than results from destructive sampling. However, separation of incipient to severe decay from benign reflectors (e.g. moisture gradient between sapwood and heartwood) in conifers was much less diagnostic than with angiosperms. A limited assessment of Acer saccharum showed that GPR has potential to detect defects in angiosperms; however, more research is needed to outline the full range of detectable defects. Based on the trees in this study, the potential for GPR to detect decay‐related defects in conifers seems limited. Despite problems detecting decay, reflections originating from the sapwood : heartwood boundary may prove useful to determine thickness of functional sapwood in conifers, but accurate quantification will require further technical development.     

Long‐term Treatment with Methylene Blue in a Dog with Hereditary Methemoglobinemia Caused by Cytochrome b5 Reductase Deficiency

A juvenile male mixed breed dog was presented for lethargy, exercise intolerance, and aggression when touched on the head. Cyanosis, tachycardia, and tachypnea were observed and persisted during oxygen supplementation. Arterial blood gas analysis by co‐oximetry identified an increased methemoglobin concentration (27%; normal, <2%) with normal arterial oxygen tension. The methemoglobinemia and associated clinical signs resolved after administration of methylene blue (1 mg/kg) IV, and the dog was discharged. The affected dog's whole‐genome sequence contained 2 potentially causal heterozygous CYB5R3 missense mutations suggesting that cytochrome b5 reductase deficiency was responsible for the methemoglobinemia. This hypothesis was confirmed by enzyme analysis that identified cytochrome b5 reductase activity in the affected dog's erythrocytes to only approximately 6% of that in a control sample. Clinical signs recurred 11 days after discharge but normalized and the methemoglobin concentration decreased with methylene blue administration PO (1.5 mg/kg, initially daily and then every other day).