Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

Suppression of feed intake after parenteral administration of D-beta-hydroxybutyrate in pygmy goats

The effect of intraperitoneal injection of D,L- or D-beta-hydroxybutyrate on feed intake and plasma metabolites was investigated in pygmy goats. The combined intraperitoneal injection of D,L-beta-hydroxybutyrate or D-beta-hydroxybutyrate (15 mmol/kg0.75) and 1,3-butanediol (6.6 mmol/kg0.75), a ketogenic substrate, decreased cumulative food intake while the same dose of 1,3-butanediol or DL-beta-hydroxybutyrate alone had no effect. The decrease in food intake after combined injection of D-beta-hydroxybutyrate and 1,3-butanediol was characterized by a significant decrease in meal frequency and a prolongation of the latency to eat. The hypophagic effect of the combined injection of D-beta-hydroxybutyrate and 1,3-butanediol was significant for 8 h, while the hypophagia after D,L-beta-hydroxybutyrate and 1,3-butanediol was significant for only 2 h after injection. Injection of D,L-beta-hydroxybutyrate increased plasma D-beta-hydroxybutyrate levels to 0.55 mmol/l and decreased plasma free fatty acids. Addition of 1,3-butanediol (6.6 mmol/kg0.75) to the injection increased plasma D-beta-hydroxybutyrate level up to 0.75 mmol/l. The results show that parenteral administration of D-hydroxybutyrate reduces feed intake in pygmy goats and that the hypophagia appears to be related to the amount of D-beta-hydroxybutyrate injected. The hypophagia seems to be related to elevated plasma D-beta-hydroxybutyrate concentration, and the threshold concentration appeared to be about 0.7 mmol/l under the experimental conditions of this study.     

Forecasting Infections of the Leaf Curl Disease on Peaches Caused by Taphrina deformans

An Israeli model forecasting leaf curl disease on peaches caused by Taphrina deformans was validated in the Emilia-Romagna region of northern Italy, during a three-year period (1996–1998), in 13 cases (year × location × cultivar). When the peach trees are susceptible to infection, the model uses mathematical functions to calculate the risk of infection on the basis of weather conditions (daily rainfall greater than 10mm, and maximum air temperature greater than 5°C), and it forecasts periods of possible symptom appearance based on the length of incubation. Peach trees became susceptible to infection between the end of January and mid March, when the first leaf buds attained phenological stage C, i.e. appearance of leaf apex. The trees remained susceptible for at least 9 weeks: the last infection occurred in mid-May.Since most of the leaf curl onsets observed in the orchards fell within the range of model forecasts, the model proved to be accurate in signalling both the first seasonal infection and repeated infections during the primary inoculum season. Few errors occurred, caused either by conditions of rainfall and temperature lower than the thresholds fixed in the model, or by discrepancies between forecast and actual length of incubation. Infection occurred also at 3.1–3.5°C, and with 9.6mm rainfall. Thus, thresholds should not be accepted too rigorously, and perhaps temperature should not be considered as a limiting factor for infection under the conditions of the present work. The length of incubation showed high variability: it was 23 days long on average, with a 95% confidence interval ranging from 20 to 27 days, and extreme values of 9 and 33 days.     

Detection of hepatitis E virus shedding in feces of pigs at different stages of production using reverse transcription-polymerase chain reaction.

The aim of this study was to determine at which production stages hepatitis E virus (HEV) is shed by the highest number of pigs and to estimate the relative risk associated with each stage. For this purpose, 146 fecal samples of pigs from 21 farms were studied. In addition, 1 sample from the manure ditch and another sample of drinking water, collected directly from the trough located in the pen, were taken from 16 farms. HEV RNA was detected in fecal samples from 34 pigs (23.29%). The production stages in which most pigs excreted HEV were weaners (41.7%) and pigs in the first month of feeding (60%). The results of the statistical analysis showed that the principal significant risk stage in HEV shedding was the first month of feeding (odds ratio [OR] 19.5, 95% CI 3.59-106.07, P = 0.001) followed by the weaners stage (OR 9.3, 95% CI .78-48.42, P = 0.008). In 8 out of 16 farms tested (50%) HEV RNA was detected in raw manure and in the water trough of only 1. Detection of HEV in manure ditches raises the concern of how to deal with manure of swine origin, because it is used as soil fertilizer.     

Electro-acupuncture and Chinese herbs for treatment of cervical intervertebral disk disease in a dog

A non-ambulatory dog with tetraparesis following a pain episode that had evolved over 2 months was submitted for medical treatment and diagnosed with intervertebral disk disease at C3-C4 and dorsal extradural compression at C1-C2 and C3-C4 using myelography and computed tomography. The dog experienced ambulation recovery after 15 days of treatment with only electroacupuncture and Chinese herbal medicine, with marked improvement occurring after only 10 treatments. Six months of follow-up demonstrated that the dog was stable and had no recurrence of symptoms. Therefore, it was concluded that the combination of electroacupuncture and Chinese herbal medicine was responsible for motor rehabilitation.     

Note: Pyrethroid resistance and possible involvement of glutathione<Emphasis Type="Italic">S</Emphasis>-transferases in<Emphasis Type="Italic">Helicoverpa armigera</Emphasis> from Turkey

An introductory study was conducted to investigate the pyrethroid resistance ofHelicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) strains in Turkey, collected from cotton fields in the Adana and Antalya provinces, through two different synthetic pyrethroid insecticides: lambda-cyhalothrin and esfenvalerate. In addition, the roles of glutathioneS-transferases (GSTs) in this resistance mechanism were analyzed. It was found that whereas resistance ratios for lambda-cyhalothrin (LD₅₀ levels) were 3- and 98-fold increased in the Adana and Antalya strains, respectively, esfenvalerate ratios were 3.3- and 92.3-fold increased in the Adana and Antalya strains, respectively, with respect to the susceptible strain. Furthermore, Adana and Antalya strains showed 2.4- and 2.9-fold higher GST activities than the susceptible strain, respectively. In the Antalya field strain, the minor increase in GST activity compared with the resistance levels implies that GSTs may be not greatly involved in this resistance. It also provides evidence that they could not be the only metabolic mechanism responsible for resistance to lambda-cyhalothrin and esfenvalerate inH. armigera from Turkey. http://www.phytoparasitica.org posting Nov. 16, 2006.     

Critical Use Exemption for methyl bromide for soil disinfestation: Italy’s experience with the European union process

Methyl bromide (MB) was phased out on January 1, 2005, and a Critical Use Exemption (CUE) process has been established in order to define the rules for allocating the needed amounts of the fumigant. This paper deals with the effects of the CUE process on Italian horticulture. The actual usage of the amounts of MB allocated to Italy for the year 2005 in the different areas and on the economically most important crops was monitored and critically evaluated. The usage pattern monitored shows that already in the first year after MB phaseout, Italy was able to replace it completely on crops such as zucchini, salad greens, basil and watermelon. However, on crops such as tomato, pepper, melon, eggplant, strawberry, and cut flowers, further work is required in order fully to replace MB. The emerging problems and research needs are discussed briefly.     

Towards identifying pathogenic determinants of the chickpea pathogen <Emphasis Type="Italic">Ascochyta rabiei</Emphasis>

Ascochyta blight is a serious disease of cool-season grain legumes (chickpea, faba bean, lentil and pea) caused by fungal species of the anamorphic genus Ascochyta and related genera. Despite extensive studies on the biology, ecology, epidemiology and management of the disease, little is known about the pathogenic determinants of these pathogens. This research aims at using Ascochyta rabiei as a model for the genus in investigating genetic factors of pathogenicity, with the ultimate goal of elucidating pathogenic mechanisms. Three advances were made: (1) insertional mutants with altered pathogenicity were identified through in vivo screening, and genomic regions adjacent to the insertion sites in selected mutants were determined; (2) a phage library of A. rabiei genomic DNA was constructed, and the library was estimated to provide complete coverage of the A. rabiei genome. This library was used successfully to recover clones with DNA adjacent to insertional mutation sites and to isolate specific genes; (3) DNA probes specific for an acyl-CoA ligase (cps1) and a polyketide synthase gene (pks1) were developed and library clones containing the corresponding genomic regions were identified from the phage library. These advances provide the foundation and necessary tools for experimentation of ectopic complementation assays and targeted mutagenesis to elucidate the genetic mechanisms of pathogenicity of A. rabiei.