Analyses of lipopolysaccharides, outer membrane proteins and DNA fingerprints reveal intraspecies diversity in Moraxella bovis isolated in Argentina

Intra-specific diversity within Moraxella bovis was investigated analysing DNA fingerprints, outer membrane proteins (OMP) and lipopolysaccharides (LPS) profiles. Three collection strains and 57 isolates of M. bovis, collected during 3 years from cattle with infectious bovine keratoconjunctivitis (IBK) symptoms, from diverse geographical locations of Argentina, were examined. The LPS and OMP profiles were studied through SDS–PAGE analysis and genotype was determined by PCR-DNA fingerprinting. Genotyping identified five DNA types while analysis of LPS and OMP profiles identified three rough LPS types and three OMP types among the 60 isolates of M. bovis including the three collection strains. None of the three methods employed to assess diversity was discriminating when used alone because the degree of heterogeneity in each group of surface structures was limited, but when data of each typing method were combined, 15 distinct subgroups were determined. This subgrouping was clearly able to differentiate isolates of the same genotype. These typing methods appear to be useful to assess different aspects of the disease such as the diversity within a population of M. bovis associated to epidemic conditions, track the causal agent in an outbreak of the disease, monitoring vaccination programs and studies on virulence.     

Genetic Diversity of Xylella fastidiosa Strains from Costa Rica, São Paulo, Brazil, and United States

ABSTRACT The diversity of 42 Xylella fastidiosa strains from Costa Rica, S?o Paulo, Brazil, and the United States were analyzed using the sequence of the 16S rRNA gene by variable number of tandem repeat (VNTR) fragment analysis and by restriction fragment length polymorphisms (RFLP) of a specific polymerase chain reaction (PCR)-amplification product using enzyme CfoI. Limited variability in the sequence of the 16S rRNA gene was observed and, although the separation was not absolute, most strains from Costa Rica clustered with strains from the United States and not with strains from S?o Paulo. The PCR-RFLP produced different patterns of DNA bands. The same pattern was shared by strains from Costa Rica, the United States, and two coffee strains from S?o Paulo, but a different pattern was observed in six coffee and orange strains from Brazil. In all, 32 amplification products were scored in the VNTR fragment analysis. The total variation observed among the X. fastidiosa strains had significant (P < 0.001) contributions from both geography and host origin as inferred by Nei's values of genetic diversity and WINAMOVA statistics. The strains from Costa Rica were isolated from diseased grapevines, coffee, and sweet orange and these strains grouped together and could be distinguished from strains from grapevine from the United States or from either coffee or sweet orange from S?o Paulo. The strains tested from Costa Rica are most likely of local origin, although the possibility that they have been introduced along with horticultural crops cannot be excluded. In either case, they are examples of independent selection of strains of X. fastidiosa affecting coffee and sweet orange. Greater genetic similarity was observed between strains from Costa Rica and the United States than with those from S?o Paulo.     

Histological Characteristics of the Equine Oviductal Mucosa at Different Reproductive Stages

The objective of this work was to study cellular changes in the epithelium of the mare’s oviduct. Oviductal samples were taken from mares at different reproductive stages for optical microscopy and Hoechst 33258 staining. Glandular-like structures were observed in 100% of the oviducts. These structures were of the tubular type and were formed by ciliated and nonciliated epithelial cells arranged in a way similar to the epithelial surface. The amount of structures decreased progressively from the ampulla to the isthmus, but did not change through the different reproductive stages. Histological changes in the epithelium of the oviduct were observed associated with the reproductive stages. In the ampulla, the amount of ciliated cells decreased in the anovulatory phase compared with other reproductive stages. Cords of connective tissue lined by epithelium (trabeculae) and dividing completely the lumen of the oviduct were found in 50% of the oviducts. Epithelial cells projected toward the lumen as large vesicles of cytoplasm, sometimes containing a nucleus. The amount of cells presenting nuclear protrusion varied throughout the oviduct, with highest incidence in the ampulla, decreasing progressively toward the isthmus (P < .05). In addition, nuclear protrusions were higher in number during the anovulatory and luteal phases than in the other reproductive stages (P < .05). These nuclear protruding cells appeared to be extruding from the epithelium and showed no signs of apoptosis based on the histological and fluorescent stains used. The existence of these gland-like structures in the oviductal mucosa should be considered when studying the oviductal physiology in mares.     

Differential interactions of virulent and non-virulent H. parasuis strains with na?ve or swine influenza virus pre-infected dendritic cells

Pigs possess a microbiota in the upper respiratory tract that includes Haemophilus parasuis. Pigs are also considered the reservoir of influenza viruses and infection with this virus commonly results in increased impact of bacterial infections, including those by H. parasuis. However, the mechanisms involved in host innate responses towards H. parasuis and their implications in a co-infection with influenza virus are unknown. Therefore, the ability of a non-virulent H. parasuis serovar 3 (SW114) and a virulent serovar 5 (Nagasaki) strains to interact with porcine bone marrow dendritic cells (poBMDC) and their modulation in a co-infection with swine influenza virus (SwIV) H3N2 was examined. At 1 hour post infection (hpi), SW114 interaction with poBMDC was higher than that of Nagasaki, while at 8 hpi both strains showed similar levels of interaction. The co-infection with H3N2 SwIV and either SW114 or Nagasaki induced higher levels of IL-1β, TNF-α, IL-6, IL-12 and IL-10 compared to mock or H3N2 SwIV infection alone. Moreover, IL-12 and IFN-α secretion differentially increased in cells co-infected with H3N2 SwIV and Nagasaki. These results pave the way for understanding the differences in the interaction of non-virulent and virulent strains of H. parasuis with the swine immune system and their modulation in a viral co-infection.     

Nutrient Losses by Runoff and Sediment from the Taluses of Orchard Terraces

The nutrient fluxes of nitrate, ammonium, phosphorus and potassium in runoff and sediments were evaluated over a two-year period (1999–2000) on the taluses of terraces, in a zone of intense subtropical orchard cultivation (SE Spain). The erosion plots were located on a terrace of 214% (65°) slope, at 180 m in altitude and with 16 m² (4 × 4 m) in area. Shrubby covers were tested for effectiveness in controlling the nutrient fluxes caused by runoff and sediments. Covers of Thymusserpylloides Bory sbsp. Gadorensis and Salvia officinalis L. reduced the NO₃ ^- runoff losses by 53 and 48%, with respect to the bare soil without natural vegetation, the NH_-4 ⁺ 61 and 56%; the PO₄ ^-3 65 and 56%; and K⁺ 58 and 46%, respectively. A greater proportion of NO₃ ^-, NH_-4 ⁺ and K⁺ were transported in runoff than in sediments. Thyme and sage with respect to the control reduced NO₃ ^- loss in sediments by 74 and 65%, NH_-4 ⁺ by 71 and 62%, P by 72 and 67%, and K by 69 and 61%, respectively. The total loss (runoff and sediments) in the bare-soil plot for NPK was 260, 39 and 888 mg m^-2 yr^-1, in the sage plot 119, 15 and 460 mg m^-2 yr^-1, and in the thyme plot 105, 12 and 360 mg m^-2 yr^-1, respectively. The results show the importance of the plant covers in soil conservation and in the recycling of nutrients on terrace slopes. This has far-reaching implications in the sense that the control of pollution from erosion is vital in reducing the eutrophication of both surface waters and groundwater located in lowlands.     

Evaluating potassium phosphonate injections for the control of <Emphasis Type="Italic">Quercus ilex</Emphasis> decline in SW Spain: implications of low soil contamination by <Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis> and low soil water content on the effectiveness of treatments

The Iberian forests are suffering severe disease and mortality as a result of decline, with Quercus ilex the major species at risk. Trunk injections with potassium phosphonate, which have been used successfully to control Phytophthora cinnamomi, were tested against decline. In an area in which P. cinnamomi was isolated, Q. ilex trees showing different degrees of decline were trunk-injected. Soil properties, and measurements of soil water content (θ) and depth to soil water table were assessed at three sites with markedly different decline incidences. Over the 5 years following the initiation of the experiment, mean symptoms among spring-treated trees and autumn-treated trees, or among trees injected twice a year (spring and autumn), once a year, and non-injected, were not significantly different. No effects of the treatments on shoot growth and acorn production were observed. However, θ values under trees which recovered from decline were higher than θ values under trees which did not recover from decline. At the site with the highest incidence of decline and tree mortality, P. cinnamomi was rarely isolated, and the presence of gravel, soil infiltration capacities and water table depth values were significantly higher than at the other sites, water stress being more likely to contribute to decline than P. cinnamomi. In areas in which θ is low, the distribution of phosphonate on the tree would be limited. Since the thresholds for phytotoxicity of potassium phosphonate in Q. ilex trees at the site studied would be higher than the amounts used, rates of the chemical slightly less than those that cause phytotoxicity should be tested.     

Low UV-C illumination for keeping overall quality of fresh-cut watermelon

The effects of four pre-packaging UV-C illumination doses (1.6, 2.8, 4.8 and 7.2 kJ m^?2) on quality changes of watermelon cubes stored up to 11 days at 5 °C were studied. Non-treated cubes were used as a control. Higher UV-C doses induced slightly higher CO₂ production throughout the storage period, while no changes in C₂H₄ production were monitored. However, UV-C did not significantly affect the final gas partial pressures within modified atmosphere packages where levels of 3–6 kPa O₂ and 13–17 kPa CO₂ were reached for all treatments. UV-C decreased microbial counts just after illumination. After 11 days at 5 °C, mesophilic, psycrophilic and enterobacteria populations were significantly lower in UV-C treated watermelon. Slight changes in CIE colour parameters were observed. According to sensory quality attributes, control and low UV-C treated cubes (1.6 and 2.8 kJ m^?2) can be stored for up to 11 days at 5 °C while the maximum shelf-life of moderate to high UV-C treated fruit was 8 days at 5 °C. Control cubes showed a 16% decrease in lycopene content after 11 days at 5 °C similar to that found for the high UV-C treatment. However low UV-C treated watermelon cubes preserved their initial lycopene content (2.8 kJ m^?2) or it was slightly decreased (1.6 kJ m^?2). UV-C radiation did not significantly affect the vitamin C content while catalase activity and total polyphenols content considerably declined throughout the storage period. However, total antioxidant capacity markedly increased, independently of UV-C doses. As a main conclusion, UV-C radiation can be considered a promising tool for keeping overall quality of fresh-cut watermelon.