Double-edged effects of the cryogenic technique for virus eradication and preservation in shallot shoot tips

Plant virus eradication is a prerequisite for the use of virus-free propagules for sustainable crop production. In contrast, virus preservation is required for all types of applied and basic research of viruses. Shoot tip cryopreservation can act as a double-edged strategy, facilitating either virus eradication or virus preservation in cryoderived plants. Here, we tested the efficacies of shoot tip cryopreservation for virus eradication and preservation in shallot (Allium cepa var. aggregatum). In vitro stock shallot shoots infected with onion yellow dwarf virus (OYDV) and shallot latent virus were thermotreated for 0, 2, and 4 weeks at a constant temperature of 36℃ before shoot tip cryopreservation. Results showed that viruses were preserved in recovered shoots when thermotherapy was not applied. Although thermotherapy lowered the regrowth levels of cryotreated shoot tips, the efficiency of virus eradication increased from 5% to 54%. Immunolocalization of OYDV and histological observation of cryotreated shoot tips showed the high frequency of virus preservation was due to the viral invasion of cells close to the apical meristem and the high proportion of cells surviving. Four weeks of thermotherapy drastically decreased the distribution of OYDV, as well as the percentage of surviving cells within the shoot tips, thereby promoting virus eradication. Virus-free plants obtained from combining thermotherapy with cryotherapy showed significantly improved vegetative growth and bulb production. The present study reports how thermotherapy can act as a trigger to facilitate either the safe preservation of Allium viruses or the production of virus-free shallot plants.     

Comparison of chlorophyll fluorescence and whole-plant bioassays of isoproturon

Summary The response to aqueous solutions of isoproturon was examined on intact plants of Matricaria perforata , Apera spica-venti and Sinapis alba in greenhouse tests, comparing visual estimates of shoot phytotoxicity at 30 d after application (DAA) with measurements of fast chlorophyll fluorescence induction at 10 DAA on leaves of the same treated plants. The sensitivity was greatest from pre-emergence application to intact plants of M. perforata and A. spica-venti , whereas post-emergence sprays were relatively more active on S. alba . The use of fluorescence induction was investigated further in Petri-dish experiments on excised M. perforata leaf segments. The Petri-dish bioassay enabled detection of isoproturon at concentrations above 9 × 10⁻⁷ M within 96 h after herbicide exposure. The potential for using chlorophyll fluorescence assays for the determination of isoproturon content in soil extracts is discussed.     

The role of organic vs. conventional farming practice,soil management and preceding crop on the incidence of stem-base pathogens on wheat

Journal of Plant Diseases and Protection - A stem-base disease complex of winter wheat includes several causal agents, such as eyespot (Oculimacula yallundae and O. acuformis), sharp eyespot...     

Growth,microtuber production and physiological metabolism in virus-free and virus-infected potato in vitro plantlets grown under NaCl-induced salt stress

Viral diseases (a biotic stress) and salinity (an abiotic stress) have been/are the two major constraints for sustainable development of the world’s agricultural production including potato. Crops grown in field are often exposed simultaneously to abiotic and biotic stress, and responses of plants to co-stress by two or more factors may differ from those to each of the multiple stresses. Using in vitro cultures, we demonstrated that virus infection (singly and in combination) or salt, and co-stress by virus infection (singly and in combination) and salt significantly reduced growth and microtuber production, and caused severely oxidative cell damage determined by levels of O₂·^? and methane dicarboxylic aldehyde, and H₂O₂ localization in situ. Alterations in physiological metabolism by increasing total soluble sugar and free proline, and by decreasing chlorophyll content are responses of potato plantlets to virus infection (singly and in combination) or salt stress and co-stress by virus infection (singly and in combination) and salt. Oxidative cell damage and reduced chlorophyll content caused by virus and/or salt are believed to be responsible for the reduced growth, eventually resulting in decreased tuber yield. Results reported here would help us to better understand possible mechanism of reduced tuber yield by virus infection and/or salt stress.     

Hematologic and biochemical reference intervals for Norwegian crossbreed grower pigs

Background: Hematologic and biochemical reference intervals depend on many factors, including environment and age. Reference intervals for Norwegian grower pigs are lacking, and previously published reference intervals for similar pigs from other countries are now outdated due to significant changes in management and breeding on the pig farms. Objectives: The aim of this study was to determine updated reference intervals for hematologic and biochemical analytes in healthy crossbred grower pigs, and to compare the results among 3 different farms. Methods: Blood samples were collected from 104 clinically healthy pigs of the most common Norwegian crossbreed (Landrace Yorkshire sow × Landrace Duroc boar). The pigs were 12–16 weeks old, weighed 30–50 kg, of both sexes, and lived on 3 farms in eastern Norway. Automated hematologic and biochemical analysis were performed using ADVIA 2120 and ADVIA 1650 analyzers. Results: Five samples were excluded because of hemolysis (1) or outliers (4). Reference intervals were calculated using parametric or nonparametric methods, depending on data distribution. Mean, median, minimum, and maximum values were tabulated. Conclusions: The reference intervals calculated in this study will be useful for the diagnosis and monitoring of disease in this widespread crossbreed pig. Compared with previously published reference values, reference intervals for total WBC count, creatine kinase and alanine aminotransferase activities, and albumin, bilirubin, and urea concentrations in this study differed notably.     

Preliminary results of intraoperative photodynamic therapy with 5-aminolevulinic acid in dogs with prostate carcinoma

Six client-owned dogs with prostate carcinoma were treated with a combination of (1) partial subcapsular prostatectomy using an Nd:YAG laser, (2) intraoperative photodynamic therapy using a halogen broad band lamp after local administration of a photosensitiser, and (3) systemic treatment with meloxicam. Median survival time was 41days (range 10-68days), which compared negatively with previous reports of subtotal laser prostatectomy combined with topical interleukin-2 administration, and photodynamic therapy alone. Despite treatment, the disease progressed locally, causing signs of stranguria to recur, and in the form of distant metastases. The recurrence of clinical signs due to the primary tumour despite photodynamic therapy is probably largely explained by insufficient penetration of light into the tissue. Better results may be obtained using other light sources (e.g. laser) and alternative techniques of light delivery, such as fibres or catheters allowing interstitial diffusion of light.     

Integration of species persistence, costs and conflicts: An evaluation of tree conservation strategies in Cambodia

Cambodia forms part of the Indo-Burma hotspot. Its extent of biodiversity, however, is subject to considerable uncertainty, as there has been little systematic collection of flora and fauna. During the Khmer Rouge regime institutions were banned, academics were prosecuted and written documentation systematically destroyed. Compared with neighbouring countries Cambodia has a low population density and relatively large natural areas that are still intact. However, deforestation is expanding rapidly and a significant but un-estimated area of forest has been degraded by development of agro-industries, encroachment, illegal logging, over-harvesting and forest fire as well as the use of chemicals during war. The purpose of the paper is to: (i) apply reserve selection methods to design more robust conservation networks when knowledge of species occurrence is incomplete and habitat is threatened, and (ii) evaluate the usefulness of systematic conservation planning in a developing country where data are limited and institutions for implementation are weak. This study investigates the performance of four non-probabilistic strategies: (i) a so-called ‘rule of thumb’, (ii) hotspot, (iii) minimum cost representation, and (iv) maximum coverage; and one probabilistic design strategy, i.e. maximum expected coverage. The maximum expected coverage approach is between 15% and 24% more efficient than the non-probabilistic strategies. Finally, the relevance of such tools to real-world conservation planning in Cambodia is investigated. By incorporating experts in the generation of data, running the models and setting up premises, they acknowledge that it is possible to contribute to more systematic conservation planning in developing countries.     

Seasonal reproduction of male Gambusia holbrooki (eastern mosquitofish) from two Florida lakes

Sixteen monthly collections of adult male Gambusia holbrooki (eastern mosquitofish) were obtained from two lakes in central Florida, USA. Lake Woodruff and Lake Apopka are 36 miles apart, but differ in several environmental parameters. Compared with Lake Woodruff, Lake Apopka is warmer, more shallow in sampling areas (particularly during drought conditions; approximately 15–90 cm in Lake Apopka versus 60–120 cm in Lake Woodruff), more turbid, and more heavily contaminated with nutrients and industrial and agricultural chemicals. Here, we present detailed information on seasonal reproduction patterns in mosquitofish in their native range and compare patterns between fish from the two lakes. Male mosquitofish were reproductively active from spring through fall. Spermatogenesis, which is regulated in part by 11-ketotestosterone, ceased in October, and fish stored spermatozoa through the winter for immediate fertilization of offspring in the spring. Compared with Lake Woodruff, fish from Lake Apopka tended to be larger and have longer gonopodia and greater gonado- and hepato-somatic indices (GSI and HSI). High GSI in Apopka fish correlated with greater spermatid production, but fewer mature spermatozoa and either the same or lower sperm counts and sperm viability. Taken together, these observations suggest that differentiation of spermatids to spermatozoa is disrupted in Apopka fish, leading to reductions in fertility in some months. Delivery of sperm to females could also be affected in Apopka fish, which exhibit lower prevalence of efferent duct tissue in the testes during the summer.     

Adverse effects of polymyxin B administration to healthy horses

BackgroundPolymyxin B (PolyB) is used to treat endotoxemia in horses; neurologic and nephrogenic adverse effects occur in humans.ObjectivesTo describe PolyB adverse effects in horses.AnimalsFive healthy horses (ataxia 0/5), 1 horse with cervical osteoarthritis (ataxia 1/5).MethodsProspective blinded randomized cross‐over trial; 3‐weeks wash out. Horses received PolyB (PolyB 6000 IU/kg IV, 7 doses q12h, n = 6) and PolyB/gentamicin (PolyB 6000 IU/kg IV, q12h 7 doses; gentamicin 10 mg/kg IV q24h 4 doses n = 4, or q12‐24 h 5 doses because of an additional erroneous dose, n = 2). Daily neurological examinations were video recorded, and ataxia graded by 3 observers. Urine status, urinary GGT/creatinine ratio, plasma creatinine, and urea were assessed every other day, EMG daily. Mixed model analysis was used to evaluate factors associated with ataxia grade and [PolyB].ResultsMedian ataxia score increased from 0/5 (range 0‐2/5) to 2/5 (range 1‐3/5) during administration and declined to 0.5/5 (range 0‐2/5) after cessation. Gentamicin co‐administration (P < .01, effect size: .8), number of PolyB doses (P < .001, effect size: .6), and time since last PolyB dose (P < .001, effect size: .5) had a significant effect on ataxia grades, while horse, day, [Genta], [PolyB], and [PolyB]_CSF did not. Gentamicin co‐administration and [Genta] C_peak had no effect on median [PolyB] C_peak (4.67 and 4.89 μg/ml for PolyB and PolyB/gentamicin, respectively). Urinary GGT/creatinine ratio was elevated in 3/6 horses receiving PolyB/gentamicin. The EMG remained unchanged.Conclusions and Clinical ImportancePolyB caused transient ataxia, worsening with cumulative PolyB doses and gentamicin co‐administration. Nephrotoxicity of PolyB was only evident when gentamicin was co‐administered.