Histopathologic patterns of nephropathy in naturally acquired canine visceral leishmaniasis

Although the nephropathy of visceral leishmaniasis (VL) is known both in humans and dogs, histopathologic alterations have not been thoroughly studied. We examined renal alterations in 55 dogs with naturally acquired VL compared with five noninfected dogs from an endemic area in northeastern Brazil. Glomerulonephritis was found in 55 dogs, interstitial alterations in 53 dogs, and tubular changes in 43 dogs with VL. The glomerular alterations found were minor glomerular abnormalities (n = 8, 14.5%), focal segmental glomerulosclerosis (n = 10, 18.2%), mesangial proliferative glomerulonephritis (n = 17, 32.7%), membranoproliferative glomerulonephritis, (n = 18, 30.9%), crescentic glomerulonephritis (n = 1, 1.8%), and chronic glomerulonephritis (n = 1, 1.8%). Morphometric and ultrastructural studies complemented the analysis. The five control animals exhibited no glomerular alterations. The glomerular lesions were related to functional alterations. Considering that the alterations of canine and human nephropathy in VL are very similar, the data obtained in this study constitute an important contribution to the understanding of canine and human VL nephropathy.     

Feed restriction followed by realimentation in prepubescent Zebu females

In the present study, the effect of restricting the feed intake for 77 days and subsequent compensatory growth for 50 days of Sindi females were evaluated. Eighteen animals with an initial age of 21 months and a mean weight of 211.7 kg were placed into three groups according to the following alimentary regimens: feed ad libitum, feed restricted to 20 % dry matter, and feed restricted to 40 % dry matter. In the feed-restriction phase, the nutrient intake decreased (P?<?0.001) with an increase in the restriction level. As a consequence, the observed decrease in ingestion and serum concentrations of total protein, albumin and globulin, urea, glucose, calcium, and phosphorus were inversely proportional (P?<?0.001) to the restriction level. Significant differences in the nutrient intake and serum concentration were not observed in the realimentation phase (P?<?0.05). When animals in the control group reached the end of the feed-restriction phase, their weights (P?<?0.05) were similar to those in the 20 % restricted group, and both obtained a final weight that was greater than that of animals in the 40 % restricted group. In the feed-restriction phase, the control group had a similar mean daily weight gain (P?>?0.05) to animals in the 20 % restricted group and (P?<?0.05) 40 % restricted group. However, in the realimentation phase, the 40 % restricted group obtained greater weight gain rates (P?<?0.05), better food conversions, and partial compensatory gains. In particular, none of the restricted groups reached the final weight of the control group. In the feed-restriction phase, ingested nitrogen, nitrogen excreted in urine and feces, nitrogen balance and retained nitrogen decreased (P?<?0.05) with an increase in the restriction level. In the realimentation phase, none of the nitrogen balance variables were influenced by the restriction level (P?<?0.05). Females in the 40 % restricted group presented better food conversion rates and greater weight gains in the realimentation phase. Based on the animals’ compensatory weight gain, a feed-restriction rate of 20 and 40 % can be adopted as a nutritional management practice for prepubescent Sindi females.     

Silicon,biological seed treatment and cutting reduce the intensity of leaf spot diseases affecting Lolium multiflorum

Italian ryegrass is a major pasture crop cultivated mainly for feeding cattle in southern Brazil. The choice of crop is based on nutritional quality and the shortage of pastures that occur during the winter in the southernmost part of Brazil. In this region, the production of biomass and seeds are threatened by diseases, in particular fungal diseases that affect the foliage. In this study we identified and monitored the natural occurrence of a complex of leaf spotting diseases and evaluated the effects of combined management practices to reduce the intensity of foliar diseases: seed treatment with Trichoderma atroviride, soil amendment with silicon and cutting frequency (none, one or two cuts). Three diseases were present in a two-season experiment: grey leaf spot (Pyricularia oryzae), brown leaf spot (Bipolaris sorokiniana) and parrot's eye leaf spot (Cercospora sp.). Grey leaf spot had the highest severity, regardless of treatment, during the first season, and was the only disease present in the following season. Grey leaf spot dominated, and plants receiving only one cut were more prone to P. oryzae infection. The treatments with silicon or T. atroviride reduced the relative disease by 40% to 80%, in relation to untreated and uncut regimes. This study showed the main leaf spot diseases associated with L. multiflorum. Moreover, it demonstrates for the first time in the region the effectiveness of reducing leaf spot via the application of silicon in the soil, inoculation of seeds with T. atroviride and managing the cutting regime.     

Plasmodium vivax and Plasmodium falciparum infection in Neotropical primates in the western Amazon,Brazil

The Brazilian Amazon is endemic for malaria and natural infections by Plasmodium spp. have been detected in Neotropical primates. Despite the diversity of primate species in the region, studies on infections by these agents are limited. The aim of the present study was to investigate the frequency of infection by Plasmodium vivax and P. falciparum in free‐born primates that were kept in captivity, in the western Amazon, Brazil. Blood samples were collected from 98 Neotropical primates. Detection of P. vivax and P. falciparum DNA was performed using a semi‐nested PCR, and the amplified products were sequenced. Plasmodium spp. DNA was detected in 6.12% (6/98) of the primates. P. vivax, and P. falciparum DNA was detected in 2.04% (2/98) and 4.08% (4/98) of these mammals, respectively. Sequencing and phylogenetic analysis confirmed the results obtained from the semi‐nested PCR. The presence of infected non‐human primates (NHP) can be auxiliary in the maintenance of P. falciparum and P. vivax and may have implications for the malaria surveillance and control in the Brazilian Amazon. It is necessary to structure an efficient surveillance system for the aetiological agents of malaria that infect NHP and humans to reduce the risk of Plasmodium spp. introduction into new areas, to protect all susceptible species.     

A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.     

Antarctic marine flora: uniquely devoid of kelps

The discovery of embryonic stages of the common large Antarctic brown seaweed Himantothallus has led to the conclusion that this plant, hitherto assigned equivocally to the Laminariales (kelps), is a member of the Desmarestiales. Moreover, field study of a large sample of Himantothallus and two other enigmatic brown algae, Phyllogigas and Phaeoglossum, has led to the merger of these three genera with the recognition of a single species, Himantothallus grandifolius. The correct placement of these kelp-like algae underscores the uniqueness of the Antarctic marine flora as the only cold-water flora without kelps.     

Validation of endogenous reference genes for expression profiling of RAW264.7 cells infected with Mycobacterium avium subsp. paratuberculosis by quantitative PCR

Reference genes are frequently used to normalize between different biological samples the levels of mRNA measured using quantitative PCR (qPCR). The expression level of many commonly used reference genes has been shown to vary between tissues or cells, or following exposure to various treatments including infection with microbes. The selection of an appropriate reference gene for an individual experiment is therefore a crucial step in the process of accurately determining changes in gene expression. For this purpose, we analyzed the expression of nine commonly used reference genes in a murine macrophage cell line, RAW264.7, for their potential use in the analysis of differential gene expression by quantitative polymerase chain reaction (qPCR) following experimental infection with Mycobacterium avium subsp. paratuberculosis. Only one of nine putative reference genes tested, casc3a, was found to be suitable, and combinations of two or more reference genes were disadvantageous. Based on data from the study, we recommend an approach for selection of reference genes, conducting assays with technical replicates in duplicate rather than triplicate, determining decision-limit quality control criteria for technical replicates and assessing the significance of gene expression fold differences using DeltaDeltaC(t) based on knowledge of the variation in the reference gene.     

Soybean cultivar performance in the presence of soybean Asian rust,in relation to chemical control programs

Soybean rust is caused by an obligate parasite (Phakopsora pachyrhizi) which has spread in Brazil in each new season since 2001 and, despite the efforts to control the disease, losses have occurred every year. Its control demands several tactics amongst which chemical control with fungicides is the main method and remains indispensable. Control strategies such as the use of cultivars with partial resistance are desirable, but are not yet commercially available. The present study analyzed the existing differences in the reactions of short, medium and long cycle soybean cultivars against Asian rust and their responses to fungicide sprays. The experiment was conducted at Uberlandia-MG, Brazil, under field conditions from December 2007 to May 2008, in the Syngenta Seeds Experimental Station. The high pressure of the disease in the experiment simulated the natural pressure that the disease often reaches in Brazil. The studied variables were: visual severity (percentage of infected leaf area), percentage defoliation and productivity (kg ha⁻¹). Disease severity was expressed as AUDPC (area under disease progress curve). Variance analysis and comparison of means by the Tukey test (5% significance) were done for all variables studied. Significant differences were observed between cultivar effects and chemical control programs. The results obtained here indicate that the cultivars M-Soy 8199RR and Emgopa 315RR were less susceptible to disease, and that a control program termed “monitoring” (in which the appearance of new pustules of the pathogen were monitored to make the decision at each fungicide spray) was the most effective.     

Leucocyte profile and growth rates as indicators of crowding stress in pejerrey fingerlings (Odontesthes bonariensis)

Crowding is one of the most common stressors found in intensive aquaculture, compromising growth rates and immune function. Plasmatic cortisol is a classic stress biomarker for fish, but its quantification is expensive and demands blood volumes that small individuals do not provide, constraining the usage of this technique to assess stress in fingerlings. The leucocyte profile is an alternative methodology to quantify stress with reduced costs and volumes of blood. Stress conditions promote neutrophilia and lymphocytopenia as response to elevated glucocorticoids levels. Considering the difficulties to assess stress imposed by intensive fish farming using measurement of glucocorticoid hormones, this study aimed to evaluate the stress‐induced changes in leucocyte profiles and growth rates imposed by crowding in fingerlings of Odontesthes bonariensis, a promising South‐American candidate for freshwater aquaculture. To meet these objectives, fingerlings (initial weight 0.05 ± 0.06 g and length 1.68 ± 0.13 cm) were reared for 45 days under three rearing densities (1, 5 and 10 fingerlings L^?1). At the end of this period, fish were anaesthetized and euthanized to obtain the leucocyte profile, neutrophil:lymphocyte ratio (N:L) and growth rate. Increasing density promoted: significant reduction in growth (final length, weight and specific growth rate); neutrophilia and lymphocytopenia; and increased N:L ratio. Concluding, the tested rearing densities imposed distinct levels of stress characterized by different N:L ratio, demonstrating that the leucocyte profile is a reliable alternative to measure stress levels in O. bonariensis fingerlings and probably in other fish species.