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Expression and localization of BCRP,MRP1 and MRP2 in intestines,liver and kidney in horse

Tydén, E., Bj?rnstr?m, H., Tjälve, H., Larsson, P. Expression and localization of BCRP, MRP1 and MRP2 in intestines, liver and kidney in horse. J. vet. Pharmacol. Therap. doi: 10.1111/j.1365‐2885.2009.01140.x. The gene and protein expression and the cellular localization of the ABC transport proteins breast cancer resistance protein (BCRP), multidrug resistance‐associated protein 1 (MRP1) and multidrug resistance‐associated protein 2 (MRP2) have been examined in the intestines, liver and kidney in horse. High gene and protein expression of BCRP and MRP2 were found in the small intestines, with cellular localization in the apical membranes of the enterocytes. In the liver, MRP2 was present in the bile canalicular membranes of the hepatocytes, whereas BCRP was localized in the cytoplasm of hepatocytes in the peripheral parts of the liver lobuli. In the kidney both BCRP and MRP2 were predominantly present in the distal tubuli and in the loops of Henle. In most tissues, the gene and protein expression of MRP1 were much lower than for BCRP and MRP2. Immunostaining of MRP1 was detectable only in the intestines and with localization in the cytoplasm of enterocytes in the caecum and colon and in the cells of serous acini of Brunner’s glands in the duodenum and the upper jejunum. The latter cells were also stained for BCRP, but not for MRP2. Many drugs used in horse are substrates for one or more of the ABC transport proteins. These transporters may therefore have important functions for oral bioavailability, distribution and excretion of substrate compounds in horse.     

Resistance to pyraclostrobin, boscalid and multiple resistance to Pristine® (pyraclostrobin + boscalid) fungicide in Alternaria alternata causing alternaria late blight of pistachios in California

Pristine® (pyraclostrobin + boscalid) is a fungicide registered for the control of alternaria late blight in pistachio. A total of 95 isolates of Alternaria alternata collected from orchards with and without a prior history of Pristine® sprays were tested for their sensitivity towards pyraclostrobin, boscalid and Pristine® in conidial germination assays. The EC₅₀ values for 35 isolates from orchards without Pristine® sprays ranged from 0·09 to 3·14 µg mL^?1 and < 0·01 to 2·04 µg mL^?1 for boscalid and Pristine®, respectively. For pyraclostrobin, 27 isolates had EC₅₀ < 0·01 µg mL^?1 and six had low resistance (mean EC₅₀ value = 4·71 µg mL^?1). Only one isolate was resistant to all three fungicides tested, with EC₅₀ > 100 µg mL^?1. Among 59 isolates from the orchard with a history of Pristine® sprays, 56 were resistant to pyraclostrobin; only two were sensitive (EC₅₀ < 0·01 µg mL^?1) and one was weakly resistant (EC₅₀ = 10 µg mL^?1). For the majority of these isolates EC₅₀ values ranged from 0·06 to 4·22 µg mL^?1 for boscalid and from 0·22 to 7·74 µg mL^?1 for Pristine®. However, seven isolates resistant to pyraclostrobin were also highly resistant to boscalid and Pristine® and remained pathogenic on pistachio treated with Pristine®. Whereas strobilurin resistance is a common occurrence in Alternaria of pistachio, this is the first report of resistance to boscalid in field isolates of phytopathogenic fungi. No cross resistance between pyraclostrobin and boscalid was detected, suggesting that Pristine® resistance appears as a case of multiple resistance.     

Risk Analysis for Latent Infection of Prune by Monilinia fructicola in California

ABSTRACT The quantitative relationships between incidence of latent infection (ILI) of prune by Monilinia fructicola and wetness duration (WD) for different bloom and fruit developmental stages and different inoculum concentrations were obtained. Three levels of ILI were considered as criteria for low, moderate, and high risks of latent infection, respectively. Seasonal patterns of WD leading to different risk levels of latent infection were obtained for low (IP(L)) and high (IP(H)) inoculum potential conditions in orchards. Longer WD was needed at a resistant than at a susceptible fruit developmental stage to induce similar levels of latent infection. The curves of WD leading to different levels of ILI over the growing season (risky WD curves) were used in risk analysis for latent infection. Multi-year historical WD data from 10 prune-growing locations in California were compared with risky WD curves. The percentage of days (P) with WD leading to a certain risk level of latent infection was calculated for each month from historical weather data. Under the IP(L) condition, the P distributions for low risk of latent infection were higher in March and April than in May and were the lowest in June for most locations. Under the IP(H) condition, the number of days that WD leading to low risk of latent infection in May increased compared with those under the IP(L) condition. The risk analysis approach was evaluated by using separate experimental data as incidence of fruit brown rot obtained from different prune orchards over years. Consistency between predicted overall risk levels of latent infection and observed incidence of fruit brown rot was obtained. The results demonstrated the usefulness of the risk analysis in decision support system for disease management.     

Factors Affecting Latent Infection of Prune Fruit by Monilinia fructicola

ABSTRACT Experiments were conducted in three prune orchards in California. In each orchard, inoculations with Monilinia fructicola, the causal agent of brown rot of stone fruits, were performed on branches of trees at bloom and fruit developmental stages. Five inoculum concentrations were used in each inoculation. Six and four wetness durations were created for each inoculum concentration at bloom and fruit developmental stages, respectively. Fruit were harvested 3 weeks before commercial harvest. The overnight freezing incubation technique was used to promote sporulation and to determine incidence of latent infection (ILI) of fruit brown rot. No differences in ILI among locations were found. A seasonal pattern of bloom and fruit susceptibility to latent infection was determined. Susceptibility to latent infection at bloom stage was at a moderate level and increased to reach the highest level at pit hardening stage. Subsequently, fruit susceptibility to latent infection decreased, reaching the lowest level in early June at embryo growth stage. Thereafter, the susceptibility increased again with fruit development and maturity until harvest. Linear relationships between ILI and inoculum concentration were obtained for most combinations of growth stage and wetness duration. Incidence of latent infection increased linearly with increased wetness duration at bloom stage and increased exponentially with increased wetness duration at early and late fruit developmental stages. The optimum temperatures for latent infection at pit hardening stage ranged from 14 to 18 degrees C, but the effect of temperature on latent infection was reduced at resistant stages. The temperature range favorable to latent infection varied for different wetness durations.     

Quantification of airborne spores of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> in stone fruit orchards of California using real-time PCR

The fungal pathogen Monilinia fructicola causes blossom blight and fruit brown rot of stone fruits in California. In this study, spore densities in the air were monitored in six orchard/year combinations with Burkard spore traps. A real-time PCR assay was developed to efficiently quantify the dynamics of spore density in these orchards during the growing season. Different patterns of dynamics of spore density were observed in these orchards. A linear relationship between numbers of spores counted with a compound microscope and those determined with the real-time PCR assay was obtained, using the same samples of spore traps. Spore density in five of six orchard/year combinations ranged from 0.0 to 0.05 spores l⁻¹, except for that in orchard 4, which showed much higher values of spore density in the air, as well as higher values and wider range of incidences of blossom infection and fruit rot than those in the other orchards. The results demonstrated a potential method to quantitatively determine spore inoculum potential in orchards by using a real-time PCR assay.     

Quantification of allele E198A in beta-tubulin conferring benzimidazole resistance in Monilinia fructicola using real-time PCR

BACKGROUND: Thiophanate-methyl, a member of the benzimidazole class of fungicides, is used in California to control brown rot of stone fruit caused by Monilinia fructicola (G. Wint.) Honey. The goal of this study was to develop a real-time polymerase chain reaction (PCR) assay as an efficient method to quantify the E198A allele of beta-tubulin that confers benzimidazole resistance. RESULTS: Using the real-time PCR assay, the frequency of allele E198A (FEA) in a population was determined from the quantities of DNA amplified with the E198A allele-specific primer pair HRF/HRR and the M. fructicola-specific primer pair MfF6/MfR6. The average proportions of highly resistant isolates determined with the conventional fungicide sensitivity method were within the range of average FEA values determined with the real-time PCR assay. We also determined the FEAs of M. fructicola populations sampled from 21 stone fruit orchards in California. Only one orchard showed a high FEA over 0.20, seven orchards had values between 0.01 and 0.1, and 13 orchards had values less than 0.01. CONCLUSION: The real-time PCR assay developed in this study provides a potentially useful tool to efficiently quantify benzimidazole resistance for large M. fructicola populations.     

Personal and family backgrounds of first-year veterinary science students at The University of Queensland

Objective To provide information on changes in the social and educational backgrounds of veterinary students over a 10 year period in an effort to determine the extent to which they are representative of the community.
Methods Questionnaires were completed by first-year veterinary students at The University of Queensland in 1985 and 1986 (152 students), and 1995 and 1996 (154), and the data were analysed using the SAS System for Windows.
Results The gender ratio of first-year veterinary students was 50:50 (male:female) in 1985 and 1986 but 10 years later it had changed to 38:62. In 1985 and 1986 77% had come directly from school, with 43% of the total coming from government schools, 17% from Catholic schools and 34% from other private (Independent) schools. A decade later the percentage coming directly from school had decreased to 40%, that from Independent schools increased to 45% and that from cities increased from 53% to 64%. The educational backgrounds of parents varied widely though a high percentage had university degrees; mothers had received less formal education than fathers, and the educational attainments of both parents were higher at the beginning than at the end of the study. More than half (57% initially; 67% 10 years later) the fathers were in professional or managerial occupations, and a similar number (50% initially; 48% 10 years later) of mothers were teachers, nurses or clerks. The number of males from country areas decreased from 26 to 16 over this period.
Conclusion These veterinary students differed from the community generally in that progressively more were female, more were from Independent schools, their parents had more formal education and more of their parents were in professional, managerial or clerical occupations.     

Ectoparasitic dermatitis in free-ranging swamp wallabies (Wallabia bicolor) in New South Wales

We report extensive dermatological lesions in three, free-ranging, adult, male swamp wallabies ( Wallabia bicolor ), presented to Taronga Western Plains Zoo wildlife hospital. All three animals were moderately infested with the louse species Heterodoxus ualabati , and two were concurrently infested with a previously undescribed sarcoptiform mite of the genus Diabolicoptes . Histological changes included moderate to marked compact hyperkeratosis, focal deep pyoderma and superficial dermatophytosis. Mild to moderate hyperplastic perivascular and periadnexal dermatitis with marked superficial and follicular hyperkeratosis was evident in the skin of the inguinal region and lateral thigh of the two wallabies infested with the Diabolicoptes species. This is the first report of ectoparasitic dermatitis in free-ranging swamp wallabies and the first report of mites of the genus Diabolicoptes from a macropodid.