Identification of potentially virulent strains of Haemophilus parasuis using a multiplex PCR for virulence-associated autotransporters (vtaA)

Haemophilus parasuis is the aetiological agent of Glässer’s disease and is also a commensal of the upper respiratory tract of pigs. Trimeric autotransporter (vtaA) genes have been identified in H. parasuis and divided into three groups on the basis of the translocator domain sequence. In this study, group 3 vtaA genes were demonstrated by PCR in all 157 H. parasuis isolates tested. Group 1 vtaA genes were associated with virulent strains; 52/54 (96%) group 1 vtaA negative field isolates were isolated from the nasal passages of healthy animals, whereas no group 1 vtaA negative field isolates were isolated from cases of Glässer’s disease. There was an association between absence of group 1 vtaA, sensitivity to phagocytosis and serum and classification of isolates into nasal cluster C by multilocus sequence typing. A multiplex PCR was developed for diagnosis of H. parasuis at the species level (group 3 vtaA positive) and to differentiate putative non-virulent strains (group 1 vtaA negative). When applied to field samples, the PCR confirmed a high prevalence of H. parasuis in conventionally farmed pigs and demonstrated that almost half of the animals carried potentially virulent strains.     

Ochratoxin A-producing fungi in Spanish wine grapes and their relationship with meteorological conditions

Forty vineyards from four wine making regions of Spain were sampled at three different growth stages in 2002 and 2003. The aim was to study the fungi associated with grapes and their ability to produce ochratoxin A (OTA) on synthetic media. Among the total mycoflora, 464 (7.7%) and 648 (10.8%) Aspergillus section Nigri (black aspergilli) strains were isolated in 2002 and 2003, respectively, and were classified into three groups: isolates with uniseriate heads, A. niger aggregate and A. carbonarius. The latter presented the highest percentage of OTA-positive strains (82% in 2002 and 76% in 2003) and produced the highest levels of toxin (2.5–25 μg g⁻¹). The sampling year, sampling date, the region and their interactions presented significant differences in the number of black aspergilli isolated. Most black aspergilli were found in 2003 and at harvest. A positive correlation between the number of black aspergilli found in grapes and the temperature in the field was found. Grapes from 2003, the warmest year, and from Costers del Segre, the warmest region, were significantly the most contaminated. No significant correlation between black aspergilli presence and other meteorological factors such as relative humidity or rainfall could be established. Musts from all the vineyards were also analysed in both years, although no OTA was found in either year.     

Polyisoprenylated benzophenones and an unusual polyisoprenylated tetracyclic xanthone from the fruits of Garcinia cambogia

In light of the wide range of biological activities of garcinol and with the aim of exploring some of them, we carried out its isolation from the fruits of Garcinia cambogia L. (Guttiferae). Surprisingly, the fruits were also found to contain guttiferones I, J, and K, compounds never reported in G. cambogia, along with three new compounds, namely, guttiferone M (1), guttiferone N (2), and the oxidized derivative of guttiferone K (6). Oxy-guttiferone K (6) is the first example of tetracyclic xanthone derived from the oxidation of a polyisoprenylated benzophenone from natural source. The natural formation of oxy-guttiferone K is in agreement with the previously described cyclization of garcinol by DPPH.     

Shelf life modeling of photosensitive food: the case of colored beverages

A new approach to shelf life modeling of photosensitive foods was developed taking into consideration the example of a saffron-containing yellow beverage highly prone to oxidative photobleaching. The beverage was exposed to different light levels at increasing temperatures. During exposure, samples were analyzed for bleaching rate, pigment content, and pigment degradation products. The results obtained clearly showed that shelf life testing of light-sensitive foods must take into proper account the effect of light. In addition, for these foods, shelf life models based on the sole accelerating effect of temperature may be misleading. By contrast, the concomitant exploitation of the accelerating effects of both light and temperature was used to develop and validate a simple model correctly predicting the shelf life of the beverage under actual storage conditions. The methodology proposed may allow solving of the difficult task of predicting shelf life of photosensitive foods usually marketed in the presence of light.     

Immuno-cytolocalization of glomalin in the mycelium of the arbuscular mycorrhizal fungus Glomus intraradices

A better understanding of the role of glomalin in arbuscular mycorrhizal fungi necessitates knowledge about the cellular functions and locations of this putative heat-shock protein (Hsp). In the present study, we determined the cellular localization of glomalin in mycelium of Glomus intraradices using immuno-electron microscopy, employing the monoclonal antibody MAb32B11. We observed that there were more gold beads bound to hyphae and spore walls than in the cytoplasm. There was also differential binding within the wall layers: L3 and L2 presented more intense labeling than the L1 layer. Our data on wall-binding of glomalin are strongly suggestive of functions other than cytoplasmic (Hsp-related), and point to a possibility of mediating interactions with the biotic and abiotic soil environment.     

Structure and fertilizer properties of byproducts formed in the synthesis of EDDHA

The synthesis of commercial EDDHA produces o,o-EDDHA as the main reaction product, together with a mixture of regioisomers (o,p-EDDHA and p,p-EDDHA) and other unknown byproducts also able to complex Fe3+. These compounds have been obtained by direct synthesis, and their structures have been determined by ESI-MS analysis as oligomeric EDDHA-like products, formed by polysubstitution in the phenolic rings. Short-term experiments show that the iron complexes of samples enriched in these oligomeric byproducts have adequate stability in solution, but a significant amount of them is lost after interaction with soils and soil materials. Mildly chlorotic cucumber plants are able to reduce iron better from o,p-EDDHA/Fe3+ than from the iron complexes of the oligomeric byproducts. In hydroponics, the chlorotic soybean susceptible plants have a lower potential for Fe absorption from these byproducts than from o,o-EDDHA/Fe3+ and from o,p-EDDHA/Fe3+. In the studied conditions, the iron chelates of EDDHA byproducts do not have the long-lasting effect shown by o,o-EDDHA/Fe3+ and present a less efficient fast-action effect than the o,p-EDDHA/Fe3+.     

Relationship between procyanidin and flavor contents of cocoa liquors from different origins

The flavor of eight cocoa liquors of different origins (Africa, America, and Asia) and different varieties (Fine grades: criollo, trinitario, and nacional. Bulk-basic grade: forastero.) was analyzed by headspace solid-phase microextraction mass spectrometry (HS-SPME-MS). Their procyanidin contents were quantified by HPLC-UV (280 nm). Fine varieties with short fermentation processes proved to contain more procyanidins, while criollo from New Guinea and forastero beans showed the highest aroma levels. The levels of cocoa aroma compounds formed during roasting are shown to vary directly with bean fermentation time and inversely with residual procyanidin content in cocoa liquor. Measurement of antioxidant activity in cocoa liquor proved to be a useful tool for assessing residual polyphenols.     

Laryngeal obstruction caused by lymphoma in an adult dairy cow

A Holstein cow was presented for inspiratory dyspnea. Endoscopic evaluation revealed swollen arytenoids and a presumptive diagnosis of bilateral arytenoidal chondritis was made. A partial arytenoidectomy was performed, the right arytenoid was submitted for histopathology, and a diagnosis of laryngeal lymphoma was made. Due to the poor prognosis, the cow was euthanized.     

Proteolysis in Hispánico cheese manufactured using a mesophilic starter,a thermophilic starter,and bacteriocin-producing Lactococcus lactis subsp. lactis INIA 415 adjunct culture

Lactococcus lactis subsp. lactis INIA 415, a strain harboring the structural genes of bacteriocins nisin Z and lacticin 481, was used as adjunct culture in the manufacture of Hispánico cheese with a mesophilic starter and a thermophilic starter of high aminopeptidase activity. Addition of the bacteriocin producer promoted early lysis of mesophilic and thermophilic starter bacteria. Extracellular aminopeptidase activity in 7-day-old cheese made using mesophilic and thermophilic starters plus bacteriocin producer was 3.0-fold the level reached in cheese made without the bacteriocin producer. Proteolysis in cheese made with mesophilic and thermophilic starters plus bacteriocin-producing adjunct culture after 25 days of ripening was 1.5-fold the level reached in cheese made without the bacteriocin producer, and the level of total free amino acids was 2.9-fold the level found in cheese made without the bacteriocin producer. Cheese made with mesophilic and thermophilic starters plus bacteriocin producer received the highest scores for flavor quality and flavor intensity and reached in 25 days the flavor intensity score of a 75-day-old cheese made without the bacteriocin producer.