Analysis of pesticide gas cartridges

Charcoal and sodium nitrate, the active ingredients in pesticide gas cartridges, are quantified via carbon analysis and ion chromatography, respectively. Linearity was excellent (R(2) > 0.995) over a range consisting of 50-150% of the target concentration for both ingredients. The coefficient of variation for the replicate analyses of gas cartridges over multiple days was <4% for both analytes. Using the results from the analysis of two batches of gas cartridges, theoretical populations were modeled and used to determine practical sampling strategies to support a quality control program for a gas cartridge manufacturing operation. This modeling indicates that the analysis of three cartridges from each of five different lots would produce mean values for both active ingredients that are within 5% of the true mean >99% of the time.     

A real-time polymerase chain reaction assay for quantification of Edwardsiella ictaluri in catfish pond water and genetic homogeneity of diagnostic case isolates from Mississippi

A quantitative polymerase chain reaction (qPCR) assay was developed for the detection and quantification of Edwardsiella ictaluri in channel catfish Ictalurus punctatus pond water using modifications to a published E. ictaluri-specific qPCR assay and previously established protocols for the molecular detection of myxozoan parasites in catfish ponds. Genomic DNA equivalents indicative of the number of bacteria in a sample were determined and standard curves correlating to bacterial numbers were established. The assay was found to be highly repeatable and reproducible, with a linear dynamic range of five orders of magnitude. There was no interference of the assay from the presence of large quantities of nontarget DNA. Known quantities of bacteria were added to sample volumes of 40 or 500 mL of pond water collected from several different ponds. The minimum level of detection was approximately 100 cell equivalents (CE) in 40 (2.5 CE/mL) or 500 mL of pond water (0.2 CE/mL). Sample volumes of 40 mL yielded the most consistent results, which were not significantly different from those obtained from broth culture alone. Cell equivalents determined by qPCR in 40-mL pond water samples spiked with known quantities of bacteria were within one order of magnitude of the actual number of cells added. Repetitive element-based polymerase chain reaction analysis of archived isolates demonstrated the genetic homogeneity of E. ictaluri, and consistent amplification of these isolates by qPCR analysis demonstrated the stability of the PCR target. The assay described here provides a reliable method for the detection and quantification of E. ictaluri in pond water and will be an invaluable tool in epidemiological studies. Additionally, the assay provides a way to evaluate the effects that vaccination, antibiotic treatments, and restricted feeding practices have on E. ictaluri populations during an outbreak. Information obtained with these tools will aid in optimizing disease management practices designed to maximize productivity while minimizing losses.     

Status of thousand cankers disease on eastern black walnut in the eastern United States at two locations over 3 years

Thousand cankers disease (TCD), a lethal fungal dieback of eastern black walnut (Juglans nigra), caused by Geosmithia morbida, and spread by the walnut twig beetle, Pityophthorus juglandis, was documented in 2009 to be very destructive in the western United States and was identified in the native range of J. nigra at Knoxville, Tennessee, in 2010, and in 2011 at Richmond, Virginia. Beginning late 2010, we studied branch dieback levels (per cent live crown) and new TCD symptom development at these two quarantined locations monthly for 3 years. Of the 106 trees studied (53 at each location), 31 trees had low live crown ratings of 70 to 0% with little change over the 3 years of the study. One per cent of the trees developed new symptoms on a per‐year basis. Thus, a moderate level of TCD (mean = 76% live crown) was present in these two locations, and most trees were in a quiescent or dormant TCD condition for 3 years, an important finding not previously reported. We found new TCD symptoms developed in Richmond in 2011 and 2012 when precipitation from January 1 to the end of August was low (60–64 cm), and not when the precipitation in Richmond was higher (99 cm). In late 2012, in Richmond, soil water potential assays indicated that some black walnut trees were under severe physiological stress (?15 bars). In contrast, in 2013, high precipitation levels (99 and 130 cm) and high soil water potentials (?0.1 to ?3 bars) at both locations were associated with extensive new foliage and stem growth and recovery from TCD. Further research is needed on water relationships in regard to TCD and black walnut health.     

Responses of leaf respiration to temperature and leaf characteristics in three deciduous tree species vary with site water availability

We measured responses of leaf respiration to temperature and leaf characteristics in three deciduous tree species (Quercus rubra L., Quercus prinus L. and Acer rubrum L.) at two sites differing in water availability within a single catchment in the Black Rock Forest, New York. The response of respiration to temperature differed significantly among the species. Acer rubrum displayed the smallest increase in respiration with increasing temperature. Corresponding Q(10) values ranged from 1.5 in A. rubrum to 2.1 in Q. prinus. Dark respiration at ambient air temperatures, expressed on a leaf area basis (Rarea), did not differ significantly between species, but it was significantly lower (P < 0.01) in trees at the wetter (lower) site than at the drier (upper) site (Q. rubra: 0.8 versus 1.1 micromol m(-2) s(-1); Q. prinus: 0.95 versus 1.2 micromol m(-2) s(-1)). In contrast, when expressed on a leaf mass basis (R(mass)), respiration rates were significantly higher (P < 0.01) in A. rubrum (12.5-14.6 micromol CO(2) kg(-1) s(-1)) than in Q. rubra (8.6-9.9 micromol CO(2) kg(-1) s(-1)) and Q. prinus (9.2-10.6 micromol CO(2) kg(-1) s(-1)) at both the lower and upper sites. Respiration on a nitrogen basis (R(N)) displayed a similar response to R(mass). The consistency in R(mass) and R(N) between sites indicates a strong coupling between factors influencing respiration and those affecting leaf characteristics. Finally, the relationships between dark respiration and A(max) differed between sites. Trees at the upper site had higher rates of leaf respiration and lower A(max) than trees at the lower site. This shift in the balance of carbon gain and loss clearly limits carbon acquisition by trees at sites of low water availability, particularly in the case of A. rubrum.     

Improving controlled pollination methodology for breeding <Emphasis Type="Italic">Acacia mangium</Emphasis> Willd.

Acacia mangium is a major plantation species for the pulp and paper industry in south-east Asia and there are a number of active breeding programs. The species is predominantly outcrossing, but with a demonstrated capacity to set selfed seed where outcross pollen is limited, with consequent inbreeding depression in the progeny. Current controlled pollination methods therefore include a time-consuming emasculation step. We used microsatellite genotyping of seedlings to determine the consequences of outcross pollination with and without emasculation. Only 1 of 3 mother trees set a small amount (5%) of selfed seed. Using whole inflorescences from the male parent as the pollen applicator rather than sieved pollen reduced outcross contamination rates from 19.1 to 8.7% and substantially increased worker productivity. Application of sugar solution to the female flowers immediately prior to pollination increased yield of sound seeds per spike. Additional improvements to the pollination protocols are discussed.     

Evict or infect? Managing populations of the large pine weevil, Hylobius abietis, using a bottom–up and top–down approach

Hylobius abietis is the most important pest of replanted coniferous sites in Northern Europe, where feeding by adult weevils can result in up to 100% mortality of seedlings. Field trials were conducted with the aim of reducing H. abietis populations developing in Sitka spruce stumps by increasing pressure from natural enemies (top–down pressure), and reducing the quality of stumps for development (bottom–up pressure). Top–down pressure was applied through inundative treatment of stumps with entomopathogenic nematodes (Heterorhabditis downesi or Steinernema carpocapsae). Bottom–up pressure was applied by treating stumps with the wood colonising fungus Trichodermakoningii. Natural levels of parasitism of H. abietis by the parasitoid Bracon hylobii and the effect of applied agents on B. hylobii were also investigated. Heterorhabditis downesi parasitised more immature weevils than S. carpocapsae, and significantly reduced numbers of adults emerging from stumps compared to controls. Entomopathogenic nematodes did not significantly impact on populations of B. hylobii, and over three sites the effects of both agents were additive. Stumps modified by application of the fungus (bottom–up pressure) did not have fewer H. abietis developing in, or emerging from them; however, development of H. abietis was more advanced in these stumps, and the success of natural enemies was differentially affected. T.koningii facilitated B. hylobii while having the opposite effect on entomopathogenic nematodes, suggesting that it affected the outcome of competition between the nematodes and the parasitoid.     

Incomplete movement of Cryphonectria hypovirus 1 within a vegetative compatibility type of Cryphonectria parasitica in natural cankers on grafted American chestnut trees

American chestnut trees, grafted in 1980 from large survivors, were inoculated in 1982 and 1983 with four white (European) hypovirulent strains of Cryphonectria parasitica, infected with C. hypovirus 1 (CHV1); this hypovirus has been shown to be capable of moving rapidly within the mycelium of a vegetative compatibility (vc) type of C. parasitica in blight cankers. Using a 49‐cell lattice plot, 17.8×17.8 cm, the spatial patterns and frequencies of white and pigmented isolates and white and pigmented vc types were investigated within superficial cankers on the grafts located outside the hypovirulent‐strain‐inoculated zone. Four of six cankers assayed contained white isolates, and three of the four had random spatial patterns of white isolates, based on join‐count statistics. Vc tests, using pigmented isolates and pigmented single‐spore colonies of white isolates, indicated that the majority of white and pigmented isolates recovered from each of two cankers assayed were in one vc type. White and pigmented lattice‐plot cells of the same vc type were frequently in contact with each other, indicating incomplete movement of CHV1 within a vc type. Nine and 10 vc types were found in the two cankers; it is hypothesized that small, white vc type areas in each canker may be a source of CHV1 transmission to the major vc types. Based on join‐count statistics, the spatial pattern of the single, major vc type in one canker was non‐random (aggregated), whereas the other canker had a random major vc type pattern. White and pigmented in vitro variants (sectors) of C. parasitica, that resemble white and pigmented in vivo variants in spatial contact and vc compatibility, were intermediate hypovirulent and virulent on forest American chestnuts, and dsRNA positive and negative, respectively. Incomplete movement of CHV1 within a vc type could be a major cause of the prevalence of pigmented isolates in superficial cankers on chestnut trees.