Autoantibody profiles in the sera of patients with Q fever: characterization of antigens by immunofluorescence,immunoblot and sequence analysis

Recent reports have shown that some of the immunological aspects of Q fever, a rickettsiosis caused by Coxiella burnetii, could be related to self-antigen responses. The aim of this study was to determine the specificity of the autoantibody response of patients with acute and chronic Coxiella infections. Smooth muscle and cardiac muscle-specific autoantibodies were observed in significant percentages in acutely or chronically affected Q fever patients when compared to healthy volunteers. Moreover, the incidence of cardiac muscle-specific autoantibody was significantly higher among chronically ill patients compared to acutely ill patients. Moreover, a band of 50 kD of a HeLa extract was detected in most of the sera of individuals with chronic infections and previous sequence analysis suggests that this antigen presents a high degree of homology with the human actin elongation factor 1 alpha. Further research would be necessary to confirm if antibodies to human cytoskeletal proteins could be of clinical importance in chronically infected Q fever patients.     

Characterization of resistance against the olive‐defoliating Verticillium dahliae pathotype in selected clones of wild olive

Verticillium wilt of olive is best managed by resistant cultivars, but those currently available show incomplete resistance to the defoliating (D) Verticillium dahliae pathotype. Moreover, these cultivars do not satisfy consumers' demand for high yields and oil quality. Highly resistant rootstocks would be of paramount importance for production of agronomically adapted and commercially desirable olive cultivars in D V. dahliae‐infested soils. In this work, resistance to D V. dahliae in wild olive clones Ac‐13, Ac‐18, OutVert and StopVert was assessed by quantifying the fungal DNA along the stem using a highly sensitive real‐time quantitative polymerase chain reaction (qPCR) protocol and a stem colonization index (SCI) based on isolation of V. dahliae following artificial inoculations under conditions highly conducive for verticillium wilt. Ac‐13, Ac‐18, OutVert and StopVert showed a symptomless reaction to D V. dahliae. The mean amount of D V. dahliaeDNA quantified in stems of the four clones ranged from 3.64 to 28.89 pg/100 ng olive DNA, which was 249 to 1537 times lower than that in susceptible Picual olive. The reduction in the quantitative stem colonization of wild olive clones by D V. dahliae was also indicated by a sharp decrease in the SCI. Overall, there was a pattern of decreasing SCI in acropetal progression along the plant axis, as well as correlation between positive reisolation and quantification of pathogen DNA. The results of this research show that wild olive clones Ac‐13, Ac‐18, OutVert and StopVert have a valuable potential as rootstocks for the management of verticillium wilt in olive.     

Investigation of biofilm production and icaA and icaD genes in Staphylococcus aureus isolated from heifers and cows with mastitis

Biofilm formation and antimicrobial resistance of Staphylococcus aureus are important virulence factors in cases of mastitis in dairy cows. However, few studies have investigated mastitis strains isolated from heifers. Within this context, the objective of the present study was to investigate biofilm formation on Congo red agar, the presence of the icaA and icaD genes by polymerase chain reaction (PCR), and the percentage of in vitro antimicrobial resistance of 110 S. aureus isolates from mammary gland secretions of heifers and cows with mastitis. PCR detected the icaA and icaD genes in 98% and 100% of isolates, respectively. However, only 55.5% of all isolates produced a biofilm on Congo red agar. Antimicrobial susceptibility testing revealed that 47.0% of isolates from heifers and 70.4% of isolates from cows were resistant to at least one of the antimicrobial agents tested. Resistance to penicillin and/or ampicillin was the most frequent (44.5%). These results indicate the need to implement prophylactic and control measures of mastitis for heifers. Heifers and cows can carry resistant strains with the capacity of biofilm production, a fact representing a threat to public health and animal well‐being and generating losses to dairy farmers.     

Morphological development and allometric growth of yellowtail kingfish Seriola lalandi V. larvae under culture conditions

Morphological development and allometric growth patterns of Seriola lalandi larvae were assessed to characterize normal growth patterns under culture conditions. Early ontogenetic stages of yellowtail kingfish exhibited an exponential growth in terms of standard length as a function of age. Five development stages were characterized from hatching to the juvenile stage: larval stage I (0–2 days post hatch, dph) with endogenous feeding, characterized by a small yolk sac, unpigmented eyes, primordial finfold surrounding the body and a closed mouth; larval stage II (2–15 dph) characterized by mouth opening, complete pigmentation of eyes and the beginning of the exogenous feeding; subsequently, in the larval stage III (15–25 dph) the posterior tip of notochord of the larvae bended upward and the first rays appeared in fins, concomitant with a change in swimming behaviour; thereafter, larval stage IV (post‐flexion stage; 25–30 dph) began when larvae resembled in morphology to a juvenile organism; however, caudal and dorsal fins were not completely development. Lastly, the juvenile stage was reached 30 dph characterized by a morphology and fin structures similar to those of the adults. Growth and development of structures and organs associated with vital functions such as feeding, sensorial and breathing systems seemed to be more critical previous to 23 dph, which was reflected with a positive allometric growth of head and eyes during this period. The results from this study can be used as a tool‐guide to assess normal development in larval research with S. lalandi to improve existing rearing protocols in hatchery production.     

Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.     

Species distribution models applied to plant species selection in forest restoration: are model predictions comparable to expert opinion?

An expert on local flora usually is the best option for plant species selection in most ecological restoration projects; although species selection often needs to be dealt with swiftly as well as on a limited budget, and obtaining the opinion of a local expert may not always be an economically viable alternative. In such cases, species distribution models (SDM) may offer a faster and more cost effective alternative. We asked six experts to rank native tree species according to their suitability at 24 forest sites. The predictive performance of the suitability rankings was evaluated by assessing their ability to discriminate present from absent species in the observed tree assemblages at each evaluation site. We used the area under the receiver operating characteristic curve to calculate the probability that the estimated suitability for a species present at a particular evaluation site is greater than the estimate for an absent species (both picked at random). Suitability rankings were also obtained from the predictions of SDM and the same procedure was used to estimate the predictive performance of the set of models at each site. The experts offered concordant suitability rankings at almost every evaluation site. There were no significant differences in the predictive performance of the SDM and four of the experts, although the SDM performed slightly better than the other two experts. Our results point to the suitability of the proposed species distribution modeling approach to obtain fast and cost effective recommendations for species selection in forest restoration projects.     

Reproductive investment of <Emphasis Type="Italic">Pinus pseudostrobus</Emphasis> along an altitudinal gradient in Western Mexico: implications of climate change

Climate varies along altitudinal gradients and species performance may be affected in response to these variations. Climate change can modify these gradients and, at the lower limit of species distributions, individuals may become stressed and their general health and reproductive performance may decrease. Study and prediction of the effects of climate change on the distribution of species along these gradients is therefore necessary. Our model study species, Pinus pseudostrobus, is a widely distributed pine in Mexico, ranging from 1600 to 3200 masl in altitude. In order to explore changes in health condition and reproductive investment along an altitudinal gradient, ten reproductive trees were selected in each of four populations (at 2300, 2400, 2700 and 2900 masl). As a proxy of individual health, we conducted a rapid visual assessment for each tree, based on crown defoliation, dead branches and foliage discoloration. This stress condition index (SCI) ranges from zero to fifteen, and lower values indicate low stress and therefore better health conditions. We also evaluated reproductive (cone weight, number/weight of seeds) and progeny (germination/seedling growth) traits. In addition, the relationships between SCI and the reproductive/progeny traits were assessed. The lowest altitude population presented poor health, with higher values of SCI (mean ± SE = 6.3 ± 0.9) that reduced up to 4.8-fold in higher-altitude populations. Cone weight also differed among altitudes, with lighter cones in the lower population (mean ± SE = 38.2 ± 4.3 g), and 1.5-fold heavier cones found in the higher populations. In general, all of the reproductive/progeny traits differed among altitudes. The population of intermediate altitude (2700 masl) presented the highest values for all traits evaluated, indicating higher performance at this altitude. Finally, a negative relationship was found between stress condition and cone weight. Based on our results and climate change models and their predictions, an increase in physiological stress can be expected in individuals of low altitude populations. Furthermore, possible increases in pests and pathogens are likely to contribute to the decline of this population. It is therefore necessary to maintain efforts of stress condition assessment and population dynamics, as well as to permanently monitor the climate along altitudinal gradients.     

Characterization of Collagen from Different Discarded Fish Species of the West Coast of the Iberian Peninsula

ABSTRACT

Skin collagen of six discarded fish species was analyzed and compared. Acid soluble collagen (ASC) was extracted; a characteristic sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profile for type I collagen was obtained, except for Chimaera mostrosa. Contents of collagen calculated from HPro (31.85% average) were higher than those determined from ASC extracts (17.75% average), with Galeus spp. being the species with the higher percentage. Amino acid analysis revealed the typical composition of collagen, with very few differences among species. Specific profiles were obtained after protease digestion. Denaturation temperature of ASC correlated well with imino and hydroxyproline contents.

Results demonstrate the feasibility of using the obtained collagens in different industrial applications.