Molecular Characterization of the USDA White Clover (Trifolium repens L.) Core Collection by RAPD Markers

White clover is one of the most important forage legume species worldwide, playing an important role in Southern Brazil temperate cultivated pastures. This work was aimed to characterize the genetic variability of the USDA white clover core collection formed by 78 accessions representing 50 countries, together with two very well known cultivars (Huia and Ladino Regal), using RAPD (Random Amplified Polymorphic DNA) markers to produce genetic fingerprints. There were used DNA bulks formed by the extraction and mixture of 20 random individuals from each accession. Twenty four primers were used, which revealed from 3 to 29 bands, forming a total of 371 polymorphic bans and only one monomorphic, ranging from 50 to 3098 bp. The results showed a genetic similarity among the accessions, ranging from 0.18 to 0.58 (Jaccard’s index), with an average of 0.24, allowing the identification of each individual accession using just three primers. The results also showed a large genetic variability within the white clover core collection, probably due to its reproduction mode and ploidy level, which could be used in plant breeding program.     

Biomass,Mineral Accumulation,and Calcium Crystals in Trema micrantha (L.) Blume as a Function of Calcium Carbonate Addition

Trema micrantha (L.) Blume is a neotropical multipurpose tree species, which could be used in forest restoration and agroforestry. Several cultures, however, require liming to satisfactory yields in acid soils. Therefore, it is important to determine the effect of liming to this native species, in order to include it in agroforestry associations. The aim of this work was to evaluate the morphological changes of T. micrantha, including the calcium oxalate (druses) and cystolith density, as caused by the calcium carbonate (CaCO₃) addition. The assay included three treatments: control, pH 5.3; treatment 1, pH 5.5; treatment 2, pH 6.0. The liming reduced the root biomass and increased the shoot biomass. The leaf specific mass and nutrients concentration were not changed. The leaves presented higher density of druses in treatment 2. The cystolith density was not modified, but the cystolith size was increased by the liming. The results indicate liming tolerance in this species.     

A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: assessment of leukocyte population changes in canine visceral leishmaniosis

A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB+ cells were higher, and of CD45RA+ cells were lower, both in the spleen and in the blood of animals with leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy.     

Radioresistance of Salmonella species and Listeria monocytogenes on minimally processed arugula (Eruca sativa Mill.): effect of irradiation on flavonoid content and acceptability of irradiated produce

This work studied the radiation resistance of Listeria monocytogenes and Salmonella species and the effect of irradiation on leaf flavonoid content and sensory acceptability of minimally processed arugula. Immersion in ozone-treated water reduced the analyzed microorganisms by 1 log. L. monocytogenes and Salmonella were not isolated from samples. Samples of this vegetable were inoculated with a cocktail of Salmonella spp. and L. monocytogenes and exposed to gamma irradiation. D10 values for Salmonella ranged from 0.16 to 0.19 kGy and for L. monocytogenes from 0.37 to 0.48 kGy. Kaempferol glycoside levels were 4 and ca. 3 times higher in samples exposed to 1 and 2 kGy, respectively, than in control samples. An increase in quercetin glycoside was also observed mainly in samples exposed to 1 kGy. In sensory evaluation, arugula had good acceptability, even after exposure to 2 and 4 kGy. These results indicate that irradiation has potential as a practical processing step to improve the safety of arugula.     

Use of NIRS for quantification of mangiferin and hesperidin contents of dried green honeybush (Cyclopia genistoides) plant material

Cyclopia genistoides, normally used for the preparation of an herbal tea, honeybush, is a good source of the bio-active compounds mangiferin and hesperidin and is in demand for the preparation of xanthone-enriched extracts. Near-infrared spectroscopy (NIRS) was used to develop calibration models to predict the mangiferin and hesperidin contents of the dried green plant material. NIRS measurements of plant material and pure compounds were performed in diffuse reflectance mode. The calibration sets for mangiferin and hesperidin contents ranged from 0.7 to 7.21 and 0.64-4.80 g/100 g, respectively. Using independent validation, it was shown that the NIRS calibration models for the prediction of mangiferin (SEP=0.46 g/100 g; R2=0.74; and RPD=1.96) and hesperidin (SEP=0.38 g/100 g; R2=0.72; and RDP=1.90) contents of the dried plant material are adequate for screening purposes, based on RPD values.     

Visceral leishmaniasis in a captive crab-eating fox Cerdocyon thous

Visceral leishmaniasis (VL) is a zoonotic disease with worldwide distribution. The crab-eating fox (Cerdocyon thous) is considered a wild reservoir of many zoonotical diseases, particularly VL. This study reported the presence of Leishmania infantum amastigotes in different organs of one captive C. thous found dead in a zoo. This animal was positive by the indirect fluorescence antibody test and had many clinical signs of VL. Intracellular amastigote forms of L. infantum were seen in neutrophils and macrophages in sample tissues from skin, lymph nodes (popliteal, submandibular, prescapular, and mesenteric), spleen, and liver. The numbers of positive cells and intracellular parasites were higher in macrophages than in neutrophils. In addition, polymerase chain reaction demonstrated extensive distribution of Leishmania DNA in C. thous tissues from multiple organs. The presence of intracellular amastigotes in neutrophils and macrophages as well as DNA of the parasite in tissues, specifically skin demonstrate that this crab-eating fox is an adequate host for L. infantum and reinforce the importance of VL for symptomatic wild canids kept in captivity in endemic areas.     

Dietary lycopene supplementation on Nile Tilapia (Oreochromis niloticus) juveniles submitted to confinement: effects on cortisol level and antioxidant response

This study evaluates the inclusion of lycopene in the commercial diet for Nile Tilapia exposured to a stressor. For 60 days, fish diet was supplemented with lycopene (600 mg kg^?1 diet) and at the end, subjected to confinement for 90 min. Zootechnical parameters (weight gain, specific growth rate and intake), hormonal (cortisol level), biochemical {enzyme activity and lipid peroxidation level [thiobarbituric acid (TBARS)]} and physical (instrumental colour) parameters were also evaluated. Supplementation with lycopene did not alter zootechnical parameters. In unconfined fish, catalase (CAT) activity was reduced in the white muscle of fish fed with lycopene; however, in red muscle, it was not altered. In fish under confinement, plasma cortisol increased as well as CAT, glutathione reductase (GR) and lactate dehydrogenase (LDH) activity in both muscles. Lipid peroxidation of the fillet also displayed higher values. However, the lycopene supplementation abolished the effects of confinement available through cortisol levels, remaining unchanged in the above parameters, activity of CAT, GR and LDH in both muscles and TBARS content of the fillet; in addition, it did not induce alteration of fillet coloration. Inclusion of lycopene in commercial feed reduces the impact of confinement and does not alter the zootechnical parameters; additionally, no modification of the fillet colour from juvenile Nile tilapia was observed.