Role of biotechnology in forest disease research and management in Canada: synthesis1

This article describes the most important biotechnological research in major tree diseases of Canadian forests and discusses the role of biotechnology in the management of forest diseases. A brief information on Canada's forest resources and the damage caused by these diseases is also included.     

Armillaria root rot: Artificial inoculation and development of the disease in greenhouse1

Inoculation of five softwood provenances have revealed that they vary considerably with regard to their percent infection, percent mortality, severity of disease symptoms and progress of infection. Sitka spruce was the most susceptible species and served as a better substrate for further development of mycelium and rhizomorphs as compared to Norway and black spruces and Scots pine. The fungus always entered through uninjured surface of root collar and/or roots, the latter with more than 2 mm diameter. There was a conspicuous reduction m the total and annual growth in height in A. mellea-infected seedlings.     

Influence of high dietary α-tocopherol intakes on specific immune response, nonspecific resistance factors and disease resistance of healthy and aflatoxin B1-induced immunocompromised Indian major carp, Labeo rohita (Hamilton)

The effect of aflatoxin treatment and/or feeding of a high level of α‐tocopherol on immune response and disease resistance was investigated in Indian major carp, Labeo rohita. Group A served as a healthy control, group B was treated with aflatoxin, group C was fed a high level of α‐tocopherol whereas group D was exposed both to aflatoxin and a high level of dietary α‐tocopherol for 60 days. Aflatoxin B₁ (AFB₁) was injected once intraperitoneally into fish on the first day of the experiment (groups B & D). High levels of DL‐α‐tocopherol (1000 mg kg^–1 feed) were provided to healthy as well as AFB₁‐treated immunocompromised fish for 60 days (groups C & D). At the end of the experiment blood samples were assayed for changes in nonspecific immunity and humoral protein levels. Disease resistance against two common bacterial pathogens viz., Aeromonas hydrophila and Edwardsiella tarda were evaluated in all groups. Significant (P < 0.05) suppression of specific immunity as measured through haemagglutination (HA) titre against sheep red blood cells (SRBCs) as well as bacterial (formalin‐killed E. tarda) agglutination titre; nonspecific resistance factors viz., globulin level, serum bactericidal and lysozyme activities, neutrophil activities, and disease resistance against two bacterial pathogens only in aflatoxin‐treated fish with respect to the control group, clearly indicated the immunosuppressive nature of aflatoxin. Feeding of a high level of α‐tocopherol to AFB₁‐treated immunocompromised fish significantly (P < 0.05) raised specific immunity, nonspecific resistance factors and disease resistance capacity when compared with aflatoxin‐exposed fish. Disease resistance and enhancement of immune status through feeding of high levels of α‐tocopherol to healthy as well as AFB₁‐treated immunocompromised fish confirmed the potential of α‐tocopherol in carp feed for prevention of disease and for combating natural/environmental immunosuppressants.     

Pharmacokinetics following intravenous administration and pharmacodynamics of cefquinome in buffalo calves

Disposition following single intravenous injection (2 mg/kg) and pharmacodynamics of cefquinome were investigated in buffalo calves 6–8 months of age. Drug levels in plasma were estimated by high-performance liquid chromatography. The plasma concentration–time profile following intravenous administration was best described by a two-compartment open model. Rapid distribution of cefquinome was evident from the short distribution half-life (t _½α ?=?0.36?±?0.01 h), and small apparent volume of distribution (Vd_area?=?0.31?±?0.008 L/kg) indicated limited drug distribution in buffalo calves. The values of area under plasma concentration–time curve, elimination half-life (t _½β ), total body clearance (Cl_B), and mean residence time were 32.9?±?0.56 μg·h/mL, 3.56?±?0.05 h, 60.9?±?1.09 mL/h/kg, and 4.24?±?0.09 h, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration of cefquinome were 0.035–0.07 and 0.05–0.09 μg/mL, respectively. A single intravenous injection of 2 mg/kg may be effective to maintain the MIC up to 12 h in buffalo calves against the pathogens for which cefquinome is indicated.     

Assessment of Genetic Diversity of Drought Tolerant and Susceptible Rice Genotypes Using Microsatellite Markers

The introgression of wild chromosomal segments into popular rice varieties is one of the potential approaches for developing varieties for drought stress condition. Sixteen genotypes, including nine indica, two tropical japonica and five chromosome segments substitution lines(CSSLs) with different levels of tolerance/susceptibility to drought stress, were selected for diversity study. Sixty-three microsatellite markers were utilized for assessing genetic diversity. A total of 95 alleles were amplified, and out of them, 60 were polymorphic. Six unique alleles, amplified by the microsatellite loci RM276, RM472,RM488, RM537, RM541 and RM28089, were identified in six genotypes, namely FR13A, Brahamanakhi,RUF44, Swarna-sub1, Brahamanakhi and Satyabhama. The highest genetic similarity was found among CSSLs. Polymorphism information content(PIC) value varied from 0 to 1.00 with an average of 0.66 per locus. Twenty-eight microsatellites were found to be polymorphic, which could be used in marker-assisted selection programme. All the sixteen genotypes were grouped into two major clusters at genetic similarity of 0.64. In the cluster I, five CSSLs identified as diverse genotypes had wild ancestor segments responsible for drought tolerance, and hence they could be utilized as potential donors. The popular Indian varieties, Swarna-sub1 and IR64-sub1, could be used as recurrent parents in the future breeding program for developing varieties for abiotic stresses such as submergence and drought.     

Remediation of Fly Ash Dumpsites Through Bioenergy Crop Plantation and Generation: A Review

Utilization of fly ash, a byproduct of coal combustion in thermal power plants, is a sustainable use of waste for power generation. Discarding fly ash as waste in landfills/ash ponds may not only be regarded as a loss of valuable land and essential nutrients, but also pose a significant health hazard due to fine air-borne particles and leaching of heavy metals. The presence of essential macro- and micronutrients and its porosity make fly ash an excellent soil amendment for plant growth as an organic nitrogen (N) and carbon (C) supplementation. As harmful heavy metals make fly ash unsafe for agronomy, bioenergy crop plantation and energy generation from different thermochemical conversions of the biomass would be an ideal method for coal fly ash utilization through which carbon-neutral fuel can be generated from fossil fuel, thus reducing climate change impact. This review summarizes the development of bioenergy plantation and silviculture at fly ash dumpsites with an integrated phyto-bio-rhizo-mycoremediation approach and assesses utilization of the valuable biomass for thermal energy, electricity, and biofuel generation with inclusion of a SWOT analysis (a strategic technique typically used to help identify the strength, weakness, opportunities, and threat). Bioenergy crop production through integrated phytomanagement can generate billions of dollars of wealth from waste and provides a sustainable solution for fly ash management, with environmental, economic, and social benefits.     

Molecular characterization and variability analysis of <Emphasis Type="Italic">Apple scar skin viroid</Emphasis> in India

Apple scar skin viroid (ASSVd) infection is a major limitation to apple fruit quality and causes huge economic losses. In surveys of apple orchards in the northern Indian state of Himachal Pradesh, fruits with dappling symptoms were noticed. ASSVd was detected from these fruits and molecularly characterized. Ten clones from three isolates were sequenced, of which seven were new sequence variants of ASSVd. The clones had significant sequence variability (94–100%) with each other. Variability was more common in the pathogenic domain of the viroid genome. Four of the clones were 330 nucleotides (nt) long, and the other six had an additional nucleotide. Phylogenetic analysis showed close affinity of the present isolates with some Chinese and Korean isolates. The study reports seven new variants of ASSVd and also provides the first molecular evidence of viroid infection (ASSVd) in apple in India.     

Pharmacokinetics of transdermal flunixin in sows

The objective of this study was to describe the pharmacokinetics (PK) of flunixin in 12 nonlactating sows following transdermal (TD) flunixin (3.33 mg/kg) and intravenous (IV; 2.20 mg/kg) flunixin meglumine (FM) administration using a crossover design with a 10‐day washout period. Blood samples were collected postadministration from sows receiving IV FM (3, 6, 10, 20, 40 min and 1, 3, 6, 12, 16, 24, 36, and 48 hr) and from sows receiving TD flunixin (10, 20, 40 min and 1, 2, 3, 4, 6, 8, 12, 16, 24, 36, 48, 60, and 72 hr). Liquid chromatography and mass spectrometry were used to determine plasma flunixin concentrations, and noncompartmental methods were used for PK analysis. The geometric mean ± SD area under the plasma concentration–time curve (AUC) following IV injection was 26,820.59 ± 9,033.88 and 511.83 ± 213.98 hr ng/ml for TD route. Mean initial plasma concentration (C₀) was 26,279.70 ± 3,610.00 ng/ml, and peak concentration (C_max) was 14.61 ± 7.85 ng/ml for IV and TD administration, respectively. The percent mean bioavailability of TD flunixin was 1.55 ± 1.00. Our results demonstrate that topical administration is not an efficient route for delivering flunixin in mature sows.     

Identification and Pathogenicity of Pseudomonas aeruginosa DJ1990 on Tail and Fin Rot Disease in Spotted Snakehead

Tail and fin rot disease (TFRD) is a big issue in the production of spotted snakehead, Channa punctata Bloch. The aims of the present study were to isolate and identify the bacterial pathogen causing TFRD, to detect histopathological changes in tissues (fin, tail, liver, and kidney), and to ascertain the antibiotic sensitivity pattern of the isolate. Out of six bacterial isolates, only the isolate DJ1990 was found to be the causal candidate of TFRD in C. punctata. Identical histopathological changes were detected in tail, fin, liver, and kidney under light and scanning electron microscope in both collected diseased fish and artificially infected fish. The isolate was identified as Pseudomonas aeruginosa strain DJ1990 (National Center for Biotechnology Information Ace. No. KX709967) based on the biochemical characterization tests and 16S rDNA sequence‐based phylogeny analysis. Artificial challenge test demonstrated that the strain DJ1990 was highly virulent (100% mortality at 48 h of postinjection period at the concentration of 1.5 × 10⁷ CFU/g of body weight) for C. punctata. The isolate exhibited sensitivity to the broad‐spectrum antibiotics but was resistant against aztreonum. To the best of our knowledge, this is the first report of P. aeruginosa as a TFRD‐causing candidate in C. punctata.