P‐20 Clinical,cytopathological and histopathological evaluation of sporotrichosis in experimentally infected cats

The purpose of this study was to evaluate the clinicopathological aspects of experimental sporotrichosis in cats and compare the sensitivity of cytopathology, histopathology and culture as diagnostic tools in different phases of the infection. Twenty adult, mixed‐breed cats (10 males and 10 females) were inoculated subcutaneously with 10⁶ fungal microorganisms. Clinical examination was performed weekly. Cytopathologic, histopathologic and culture examinations were performed at 15, 30 and 60 days postinoculation. Culture of multiple organs was performed after euthanasia at 30 (10 cats) and 60 (10 cats) days postinoculation. Friedman parametric and nonparametric statistical analysis were applied to the results. The nodular, tumoral and necrotic lesionsprogressed significantly until day 30 postinoculation, and partial spontaneous regression occurred at day 60, particularly in males. An intense inflammatory pyogranulatomous and lymphocytic infiltrate with rare giant cells and sparse fibrosis associated with numerous, pleomorphic, intra‐ and extracellular fungal cells were observed on day 30. These findings gradually decreased by day 60. Despite the inflammatory granuloma associated with feline sporotrichosis, a tendency for dissemination was observed, with fungal isolation in the lymph nodes, spleen and liver at the 30 and 60 days postinoculation. No significant differences were observed between cytopathology, histopathology and fungal culture during the different phases of the disease. Therefore, cytological examination was considered a simple, rapid and inexpensive diagnostic method at all stages of this disease. Funding: Self‐funded.     

Effect of consumption of organically and conventionally produced apples on antioxidant activity and DNA damage in humans

The present study was performed to compare the effects on antioxidant activity and on DNA damage of organic and conventionally produced apples grown under controlled conditions in human peripheral blood lymphocytes. Six healthy volunteers consumed either organically or conventionally grown apples (Golden Delicious, 1000 g) from two neighboring commercial farms in a double-blinded, randomized, cross-over study. The average content of total identified and quantified polyphenols in the organically and conventionally produced apples was 308 and 321 microg/g fresh weight, respectively. No statistically significant differences in the sum of phenolic compounds or in either of the polyphenol classes were found between the agricultural methods. Consumption of neither organically nor conventionally grown apples caused any changes in antioxidant capacity of low-density lipoproteins (lag time test), endogenous DNA strand breaks, Fpg protein-sensitive sites, or capacity to protect DNA against damage caused by hydrogen peroxide. However, a statistically significant decrease in the levels of endonuclease III sensitive sites and an increased capacity to protect DNA against damage induced by iron chloride were determined 24 h after consumption in both groups of either organic or conventionally grown apples, indicating the similar antigenotoxic potential of both organically and conventionally grown apples.     

Lipoxygenase distribution in coffee (Coffea arabica L.) berries

In this paper lipoxygenase (LOX) presence was investigated in coffee berries to determine its involvement in lipid degradative metabolism of plants grown in organic and conventional cultivations. An immunochemical analysis has evidenced a ca. 80 kDa protein, cross-reacting with an anti-LOX antibody, only in the pulp fraction of berries obtained from plants of both cultivations. LOX activity in this fraction could be monitored either as conjugated diene formation or reaction products (determined by HPLC) and was mainly associated with a heavy membrane fraction (HMF, enriched in tonoplast, endoplasmic reticulum, plasma membrane, and mitochondria) and a light membrane fraction (LMF, enriched in plasma membrane and endoplasmic reticulum, with low levels of tonoplast and mitochondria). The LOX activity of LMF from berries of both cultivations showed an optimum at pH 8.0. The HMF exhibited a different activity peak in samples from conventional (pH 8.0) and organic (pH 5.5) cultures, suggesting the presence of different isoenzymes. These findings were also confirmed by variation of the ratio of 9- and 13-hydroperoxides in organic (1:1) and conventional cultivations (1:10), indicating that the organic one was subjected to an oxidative stress in the coffee pulp fraction leading to the expression of an acidic LOX. Such de novo synthesized LOX activity could be responsible for the production of secondary metabolites, which may interfere with the organoleptic profile of coffee.     

Hemangiosarcoma of the third eyelid in a cat

A case of hemangiosarcoma of the third eyelid in a 15-year-old Domestic Short-haired cat is reported. A small red mass protruding from the anterior surface of the third eyelid in the left eye was present. Only mild bilateral conjunctivitis and nuclear sclerosis were revealed on initial ophthalmic examination. After anti-inflammatory treatment, surgery and cryotherapy were performed. On histopathologic examination the mass was located just beneath the conjunctival epithelium, suggesting that the conjunctiva was the tissue of origin. The tumor showed high cellularity and was composed of pleomorphic spindle and polygonal cells arranged in interlacing bundles and solid sheets. The presence of slit-like spaces between tumor cells containing erythrocytes demonstrated the vascular differentiation. Mitotic activity was moderate, but atypical mitoses were detected. The surgical area healed uneventfully and after 7 months there was no recurrence. To the authors' knowledge, this is the first report of conjunctival hemangiosarcoma of the third eyelid in a cat.     

Removal of nutrient limitations by long-term fertilization decreases nocturnal water loss in savanna trees

Under certain environmental conditions, nocturnal transpiration can be relatively high in temperate and tropical woody species. We have previously shown that nocturnal sap flow accounts for up to 28% of total daily transpiration in woody species growing in a nutrient-poor Brazilian Cerrado ecosystem. In the present study, we assessed the effect of increased nutrient supply on nocturnal transpiration in three dominant Cerrado tree species to explore the hypothesis that, in nutrient-poor systems, continued transpiration at night may enhance delivery of nutrients to root-absorbing surfaces. We compared nocturnal transpiration of trees growing in unfertilized plots and plots to which nitrogen (N) and phosphorus (P) had been added twice yearly from 1998 to 2005. Three independent indicators of nocturnal transpiration were evaluated: sap flow in terminal branches, stomatal conductance (g(s)), and disequilibrium in water potential between covered and exposed leaves (DeltaPsi(L)). In the unfertilized trees, about 25% of the total daily sap flow occurred at night. Nocturnal sap flow was consistently lower in the N- and P-fertilized trees, significantly so in trees in the N treatment. Similarly, nocturnal g(s) was consistently lower in fertilized trees than in unfertilized trees where it sometimes reached values of 150 mmol m(-2) s(-1) by the end of the dark period. Predawn gs and the percentage of nocturnal sap flow were linearly related. Nocturnal DeltaPsi(L) was significantly greater in the unfertilized trees than in N- and P-fertilized trees. The absolute magnitude of DeltaPsi(L) increased linearly with the percentage of nocturnal sap flow. These results are consistent with the idea that enhancing nutrient uptake by allowing additional transpiration to occur at night when evaporative demand is lower may avoid excessive dehydration associated with increased stomatal opening during the day when evaporative demand is high.     

Phenotypic differences may limit the potential for habitat restoration involving species translocation: a case study of shape ecophenotypes in different populations of Modiolus modiolus (Mollusca: Bivalvia)

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Multiply-primed rolling-circle amplification (MPRCA) of PCV2 genomes: Applications on detection, sequencing and virus isolation

Multiply-primed rolling-circle amplification (MPRCA) was used to amplify porcine circovirus type 2 (PCV2) genomes isolated from tissues of pigs with signs of post-weaning multisystemic wasting syndrome (PMWS). Two of the amplified PCV2 genomes were cloned in prokaryotic plasmids and sequenced. Both were nearly identical (1767 nt) except for one silent substitution in the region coding for the capsid protein (ORF2). In addition, they showed high nucleotide sequence similarity with PCV2 isolates from others countries (93–99%). To investigate whether the MPRCA amplified PCV2 genomes could be used to produce infectious virus, the cloned genomes were isolated from the plasmids, recircularized and used for transfection in PK-15 cells. This procedure led to the production of infectious virus to titres up to 10^5.55 TCID₅₀/mL. It was concluded that MPRCA is a useful tool to amplify PCV2 genomes aiming at sequencing and virus isolation strategies, where particularly useful is the fact that it allows straightforward construction of PCV2 infectious clones from amplified genomes. However, it was less sensitive than PCR for diagnostic purposes.