Pyrazinamide inhibits trans-translation in Mycobacterium tuberculosis

Pyrazinamide (PZA) is a first-line tuberculosis drug that plays a unique role in shortening the duration of tuberculosis chemotherapy. PZA is hydrolyzed intracellularly to pyrazinoic acid (POA) by pyrazinamidase (PZase, encoded by pncA), an enzyme frequently lost in PZA-resistant strains, but the target of POA in Mycobacterium tuberculosis has remained elusive. Here, we identify a previously unknown target of POA as the ribosomal protein S1 (RpsA), a vital protein involved in protein translation and the ribosome-sparing process of trans-translation. Three PZA-resistant clinical isolates without pncA mutation harbored RpsA mutations. RpsA overexpression conferred increased PZA resistance, and we confirmed that POA bound to RpsA (but not a clinically identified ΔAla mutant) and subsequently inhibited trans-translation rather than canonical translation. Trans-translation is essential for freeing scarce ribosomes in nonreplicating organisms, and its inhibition may explain the ability of PZA to eradicate persisting organisms.     

Sensitivity of Direct Culture,Enrichment and PCR for Detection of Campylobacter jejuni and C. coli in Broiler Flocks at Slaughter

Broiler chicken flocks are a significant source of Campylobacter jejuni and Campylobacter coli that result in the major public health problem of campylobacteriosis. Accurate estimates of the prevalence of both C. coli and C. jejuni in flocks would enhance epidemiological understanding, risk assessment and control options. This study combined results from a panel of 10 detection tests (direct culture, enrichment and PCR) on caecal samples from flocks at slaughter. A parallel interpretation approach was used to determine the presence of Campylobacter spp. and for C. jejuni and C. coli individually. The sample was considered positive if at least one method detected the target and this interpretation was taken to represent a ‘proxy gold standard’ for detection in the absence of a gold standard reference test. The sensitivity of each individual method to detect Campylobacter spp., C. jejuni and C. coli was then estimated relative to the proxy gold standard. Enrichment in adapted Exeter broth (deficient in polymyxin B) with a resuscitation step was 100% sensitive, whilst direct culture on modified charcoal cefoperazone deoxycholate agar (mCCDA) was highly sensitive (97.9%). Enrichment methods using Preston broth and Bolton broth were significantly less sensitive. Enrichment in Exeter broth promoted the recovery of C. jejuni, whilst enrichment in Bolton broth favoured C. coli. A RT‐PCR detection test could identify 80% of flocks that were co‐colonised with both species. This study found that 76.3% (n = 127) of flocks were colonised with Campylobacter spp. The majority (95.9%) of Campylobacter‐positive flocks were colonised with C. jejuni; however, approximately one‐third of positive flocks were simultaneously colonised with both C. jejuni and C. coli. The findings highlight the impact of different detection methodologies on the accuracy of the estimated incidence of both C. jejuni and C. coli entering the abattoir within broiler flocks and the associated public health risks.     

Incidence rate of musculoskeletal injuries and determinants of time to recovery in young Australian Thoroughbred racehorses

Objective To describe the incidence rate of musculoskeletal (MS) injuries and investigate the factors that influence recovery from MS injury in young Thoroughbred horses in Australia. Methods Training and injury data were collected at fortnightly intervals from 14 trainers in New South Wales. Incidence rates were calculated for initial injuries stratified by age‐class and sex. Survival analysis methods were used to investigate the time to recovery after the first MS injury (measured as a start in a race or barrier trial). Results For the 248 racehorses enrolled, there were 428 injuries recorded; 2‐year‐olds were 2.99‐fold more likely than 3‐year‐olds to sustain an MS injury. The cumulative percentage of horses that had recovered within 6 months was 55%. The time to recovery was longest in horses whose fast gallop speed prior to injury was < 890 m/min. Conclusion This study supports the finding that shin soreness is the most common MS injury in young Australian Thoroughbreds. There was an increased rate of MS injury in 2‐year‐olds compared with 3‐year‐olds. However, it was impossible to determine if the increased risk in 2‐year‐olds was related to age, the start of training or a combination of both. The study also found that horses performing at higher exercise speeds prior to injury were more likely to recover. There was a significant effect of trainer, suggesting that other factors related to trainer may influence the time to the recovery. Additional research is required to understand the determinants of time to recovery.     

Developmental decline in height growth in Douglas-fir

The characteristic decline in height growth that occurs over a tree's lifespan is often called "age-related decline." But is the reduction in height growth in aging trees a function of age or of size? We grafted shoot tips across different ages and sizes of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees to determine whether the decline in height growth is mediated by tree size or by the age of the apical meristem. We also evaluated whether reduced carbon assimilation plays an important role in height growth decline. In one experiment we cut shoot tips from old-growth, young-mature and seedling trees and grafted them onto 2-year-old graft-compatible rootstock in a seed orchard in Lebanon, Oregon. In another experiment we performed reciprocal grafts between lateral branches of old-growth trees accessible from the canopy crane at Wind River, Washington and young-mature trees in a nearby plantation. We measured growth (diameter and elongation of the dominant new stem) and mortality annually for three years in the Seed Orchard experiment and for two years in the Reciprocal Graft experiment. In the Seed Orchard experiment we also measured photosynthetic capacity (determined from the response of net carbon assimilation to the intercellular CO(2) concentration of the leaf, or A/C(i) curves), leaf mass per area (LMA) and carbon isotope composition (delta(13)C) of cellulose in 1-year-old foliage. Grafting caused changes in both growth and physiology of the grafted stems. Within two years after grafting, growth and physiology of all combinations of scions and rootstock exhibited characteristics of the rootstock. In some cases, the change in growth was dramatic-cuttings from old-growth trees showed a 10-fold increase in stem elongation rate within 2 years of grafting onto seedling rootstock. Similarly, carbon isotope discrimination of new foliage on shoots from old-growth trees increased by nearly 3 per thousand and 2 per thousand after grafting onto young-mature and seedling rootstock, respectively, whereas discrimination decreased by a similar magnitude in scions from young-mature trees after grafting on old- growth trees. Furthermore, differences in carbon assimilation estimated from carbon isotope discrimination and A/C(i )relationships were small relative to growth differences. Our results confirm that size, not age, drives developmental changes in height growth in Douglas-fir. Reduced carbon assimilation does not play an important role in height growth decline.     

Controlling nitrogen release from farm ponds with a subsurface outflow device: Implications for improved water quality in receiving streams

The retention of nutrients in farm ponds has many potential benefits, including reduction of nitrogen and phosphorus (promoters of eutrophication) in receiving streams. The aim of this study was to evaluate the efficacy of a commercial subsurface pond outflow control device (Pond Management System™) on nutrient retention in farm ponds. Four ponds of similar size and water chemistry in the upper Tar River basin of North Carolina, USA were studied; three were equipped with the pond outflow control device and one was retained without a device (normal surface outflow) that served as a reference site. Water samples were collected monthly from each pond at 0.3 m intervals from the surface to 2.1 m at a fixed station adjacent to the pond standpipe and from the pond outflow pipe from March to October 2005. The water samples were analyzed for total Kjeldahl nitrogen (N), total phosphorus (P), chlorophyll a, and a suite of other physicochemical variables. In ponds with the subsurface outflow device, the mean N concentrations in the outflow were substantially less (6.2–20.7%) than concentrations at the pond surface. Concentrations of N in the outflow were similar to N concentrations at intermediate pond depths (0.9–1.5 m), the depth of the outflow devices, indicating water was drawn from these depths and that N was being retained in the surface layers of the pond. Also, mean water temperatures were 1.1–1.9 °C cooler at intermediate depths compared to the surface, suggesting potential application of the outflow device for minimizing warm water outflows to receiving streams. These results provide evidence that under these conditions a subsurface pond outflow device can reduce nutrient release to receiving streams, thereby increasing overall stream water quality.