Proposal to reclassify Paenibacillus larvae subsp. pulvifaciens DSM 3615 (ATCC 49843) as Paenibacillus larvae subsp. larvae. Results of a comparative biochemical and genetic study

The bacterial pathogen Paenibacillus larvae subsp. larvae (P. l. larvae), is the etiological agent of American foulbrood, an extremely contagious and disastrous disease of honeybee brood. In case of American foulbrood the destruction of infected colonies is often considered the only workable control measure. Therefore, the ability to diagnose this disease properly is important to prevent unnecessary economic loss to beekeepers. The development of suitable methods for the early and reliable detection of P. l. larvae is hampered by the fact that the two subspecies of Paenibacillus larvae, P. l. larvae and Paenibacillus larvae subsp. pulvifaciens (P. l. pulvifaciens), seem to be indistinguishable by cultural characteristics as well as by PCR protocols. Here we present an extensive analysis of several P. larvae reference strains. We employed conventional culture techniques, morphological and biochemical identification, PCR-based methods and sequencing of the 16S rDNA. We found indeed that P. l. pulvifaciens strain DSM 3615 is indistinguishable from P. l. larvae (DSM 7030). We did not face any problems to discriminate between P. l. larvae and P. l. pulvifaciens strains DSM 8442 and DSM 8443. Therefore, classification of DSM 3615 as type strain of P. l. pulvifaciens seems not to be justified. We propose to reclassify this strain as P. l. larvae. Former problems in differentiating the two subspecies might have arisen from this misclassification. PCR-based methods as well as appropriate biochemical identification systems provide a reliable means for the discrimination between the two subspecies P. l. larvae and P. l. pulvifaciens.     

Emergency presentations of 4 dogs with suspected neurologic toxoplasmosis

Objective: To review the signalment, clinical signs, abnormal laboratory data, therapeutics, and response to therapy of dogs with clinical signs consistent with toxoplasmosis infection. Series summary: A retrospective review was performed on the records of 4 dogs presented to the Animal Emergency Center between January 1998 and February 2000 exhibiting neurologic signs and having elevated titers for Toxoplasma gondii. A tentative diagnosis of toxoplasmosis was based upon one of the following criteria: (1) a serial 4‐fold or greater change in serum T. gondii IgG titers; 2) serially decreasing serum T. gondii IgM titers with concurrent increasing serum T. gondii IgG titers; or 3) positive cerebrospinal fluid (CSF) T. gondii titers. In addition, inclusion of cases was limited to dogs that showed improvement of neurologic signs following treatment with antiprotozoal drugs. Trimethoprim–sulfamethoxazole treatment was associated with successful elimination of clinical signs in all of the dogs. Two of the dogs developed side effects potentially attributed to the trimethoprim–sulfamethoxazole (TMS), and antiprotozoal treatment was continued using clindamycin. Unique information presented: Toxoplasmosis is an important differential diagnosis in any dog that presents as an emergency with central or peripheral neurologic signs. Affected dogs need not be immunocompromised for clinical signs of toxoplasmosis to occur. Appropriate treatment with TMS or clindamycin can lead to resolution of clinical signs.     

Comparison of Two Methods For Determining the Discoloration Potential of Potato Tubers Based on Their Chemical and Biochemical Properties

The aim of this study was to compare the homogenization and visual methods that are used to determine the discoloration potential of potato tubers. Two German cultivars, Agria and Satina, and two Polish cultivars, Lena and Mors, were planted during 1998 and 1999 in Goettingen, Germany, and in Bydgoszcz, Poland. The discoloration potential of the tubers was measured after harvesting and after 6 months of storage at 4 C and 8 C. Both methods correlated significantly at harvest (0.709,P < 0.001). After storage the correlation was significant but low (r = 0.403,P < 0.001) probably because of physical changes in the tuber during storage. The visual method can be used as a fast method, especially at harvest. However, the homogenization and visual methods determine, to a certain degree, different tuber properties leading to the discoloration potential. Therefore, a complete comparison is not possible. The methods studied in this paper can complement but not substitute each other. Reliable comparisons between institutes require use of the same method to determine the discoloration potential of the tuber.     

Quantifying rice yield gaps and their causes in Eastern and Southern Africa

The demand for rice in Eastern and Southern Africa is rapidly increasing because of changes in consumer preferences and urbanization. However, local rice production lags behind consumption, mainly due to low yield levels. In order to set priorities for research and development aimed at improving rice productivity, there is a need to characterize the rice production environments, to quantify rice yield gaps—that is, the difference between average on-farm yield and the best farmers’ yield—and to identify causes of yield gaps. Such information will help identifying and targeting technologies to alleviate the main constraints, and consequently to reduce existing yield gaps. Yield gap surveys were conducted on 357 rice farms at eight sites (19–50 farmers per site) across five rice-producing countries in Eastern and Southern Africa—that is Ethiopia, Madagascar, Rwanda, Tanzania and Uganda—for one or two years (2012–13) to collect both quantitative and qualitative data at field and farm level. Average farm yields measured at the eight sites ranged from 1.8 to 4.3 t/ha and the average yield gap ranged from 0.8 to 3.4 t/ha. Across rice-growing environments, major causes for yield variability were straw management, weeding frequency, growth duration of the variety, weed cover, fertilizer (mineral and organic) application frequency, levelling and iron toxicity. Land levelling increased the yield by 0.74 t/ha, bird control increased the yield by 1.44 t/ha, and sub-optimal management of weeds reduced the yield by 3.6 to 4.4 t/ha. There is great potential to reduce the current rice yield gap in ESA, by focusing on improvements of those crop management practices that address the main site-specific causes for sub-optimal yields.     

Von dauerbrache und fruchtfolge auf die n‐bilanzen einer löss‐schwarzerde unter berücksichtigung extremer düngungsvarianten

In einem zweifaktoriellen Dauerversuch (Anlage 1983/84) auf Löß‐Schwarzerde im Mitteldeutschen Trockengebiet wurde der Einfluß steigender Stalldunggaben bis zu 200t/ha.a auf die N‐Bilanz und die Verlagerung mineralischen Stickstoffs in tiefere Bodenschichten (bis 5 m Tiefe) in einer Fruchtfolge und unter Dauerschwarzbrache untersucht. Ein beträchtlicher Teil des Stickstoffs wurde in 0–60 cm Tiefe in der organischen Substanz des Bodens akkumuliert. Der mineralische Stickstoff ist mit Pflanzenbewuchs bis in eine Tiefe von 0–100 cm, unter Schwarzbrache bis ca. 3m Tiefe zu finden. Die Gefahr hoher gasförmiger N‐Verluste (im vorliegenden Versuch ca. 20%) bzw. von Auswaschungsverlusten beim Brachlegen von Ackerflächen wird durch diese Untersuchungen unterstrichen.     

Purification and thermal and high-pressure inactivation of pectinmethylesterase isoenzymes from tomatoes (Lycopersicon esculentum): a novel pressure labile isoenzyme

Tomato pectinmethylesterase (PME) was successfully purified by a two-step method consisting of affinity chromatography followed by cation exchange chromatography. According to this procedure, four different isoenzymes were identified representing molar masses around 34.5-35.0 kDa. Thermal and high-pressure inactivation kinetics of the two major isoenzymes of tomato PME were studied. A striking difference between their process stability was found. The thermostable isoenzyme was completely inactivated after 5.0 min at 70 degrees C, whereas for the thermolabile isoenzyme, temperatures at around 60 degrees C were sufficient for complete inactivation. The thermostable isoenzyme was also found to be pressure stable since no inactivation was observed after 5.0 min of treatment at 800 MPa and 20 or 40 degrees C. The thermolabile isoenzyme appeared to be pressure labile since it could be completely inactivated after 5.0 min of treatment at 700 MPa and 20 degrees C or 650 MPa and 40 degrees C. Inactivation kinetics at pH 6.0 could be accurately described by a first-order model.     

Monocyclic Aromatic Hydrocarbons in Kathmandu During the Winter Season

Mixing ratios of seven monocyclic aromatic hydrocarbons, as well as NO₂, SO₂ and O₃, were measured by long path differential optical absorption spectroscopy (DOAS) at a suburban site in Kathmandu, Nepal, during Jan.–Feb. 2003. The results showed average benzene (3.9?±?1.8 ppbv), toluene (13.3?±?7.1 ppbv), and sum of xylene isomers (42.2?±?15.7 ppbv) mixing ratios in Kathmandu. The xylenes concentrations were higher than in the large cities that have been studied. The observed ratio of toluene to benzene (2.9?±?1.8) reflected the small fraction of vehicles with catalytic converters in the Kathamndu. Analysis of the late afternoon time series of aromatics, NO _x , and wind data indicated that road traffic was one of the main sources of aromatics in the urban air. In addition, the correlations between aromatics, SO₂, NO _x , and PM₁₀ during the night strongly suggested that fossil and biomass fuel burning made an important contribution to air pollution in the Kathmandu valley. Aromatic pollution was further strengthened by daily recurring stable meteorological conditions and the surrounding topography. The chemical reaction of aromatics with free radicals during the daytime could also be deduced. High ratios of phenol/benzene and para-cresol/toluene could not be explained by chemical processes, and suggested direct emission of phenol and para-cresol in the Kathmandu atmosphere.     

Analysis of Genetic Variation Among Accessions of <Emphasis Type="Italic">Pueraria montana</Emphasis> (Lour.) Merr. var. <Emphasis Type="Italic">lobata</Emphasis> and <Emphasis Type="Italic">Pueraria phaseoloides</Emphasis> (Roxb.) Benth. based on RAPD Markers

Pueraria montana var. lobata and P. phaseoloides originating from tropical Asia and parts of Oceania are ecologically and economically important legumes that are used as green manure, cover crop or forage plants. Conservation and use of plant genetic resources require an understanding of the extent and distribution of genetic diversity in any given region. In this study, genetic variation of five P. montana var. lobata and 16 P. phaseoloides accessions was analysed developing a random amplified polymorphic DNA (RAPD) marker methodology for Pueraria species and thereby creating basic data for follow-up research and the development of conservation strategies. Seeds were collected from native populations in Bac Kan Province, a mountainous region in Northeast Vietnam. P. montana var. lobata presented a high level of variation with 54.3% of the detected markers being polymorphic, whereas P. phaseoloides exhibited an intermediate to high level of variation (45.5%). The P. montana var. lobata accessions clustered in congruence with their eco-geographical origin. For P. phaseoloides no correspondence between sampling sites and genetic differentiation was found. Inter-population differentiation was measured as Jaccard's similarity coefficient (JSC). Mean JSC amounted to 0.35 in P. montana var. lobata and 0.52 in P. phaseoloides. Results are compared to other genetic studies of herbaceous legumes and conservation strategies are suggested.     

Modifications of strawberry fruit antioxidant pools and fruit quality under NaCl stress

The responses of fruit antioxidants in two strawberry cultivars differing in their sensitivity to NaCl stress were studied. The sensitive cv. Elsanta and the less sensitive cv. Korona were treated with NaCl solutions of 40 and 80 mmol/L in the nutrient solution. In general, moderate salinity resulted in increases of antioxidant capacity. In cv. Korona, salt stress increased the activity of superoxide dismutase and the contents of glutathione, phenols, and anthocyanins, while that of ascorbic acid decreased. In cv. Elsanta, changes of superoxid dismutase activity and of ascorbic acid concentration were comparable to those of cv. Korona, but the decrease of ascorbic acid was more distinct. The contents of anthocyanins decreased, and those of phenols remained similar. The glutathione content was reduced at the highest NaCl level. The results indicate that less salt-sensitive strawberry cultivars may be grown under moderate salinity stress to optimize fruit quality.