Comparative analyses of whole genome sequences of Leishmania infantum isolates from humans and dogs in northeastern Brazil.

The genomic sequences of 20 Leishmania infantum isolates collected in northeastern Brazil were compared with each other and with the available genomic sequences of 29 L. infantum/donovani isolates from Nepal and Turkey. The Brazilian isolates were obtained in the early 1990s or since 2009 from patients with visceral or non-ulcerating cutaneous leishmaniasis, asymptomatic humans, or dogs with visceral leishmaniasis. Two isolates were from the blood and bone marrow of the same visceral leishmaniasis patient. All 20 genomic sequences display 99.95% identity with each other and slightly less identity with a reference L. infantum genome from a Spanish isolate. Despite the high identity, analysis of individual differences among the 32 million base pair genomes showed sufficient variation to allow the isolates to be clustered based on the primary sequence. A major source of variation detected was in chromosome somy, with only four of the 36 chromosomes being predominantly disomic in all 49 isolates examined. In contrast, chromosome 31 was predominantly tetrasomic/pentasomic, consistent with its regions of synteny on two different disomic chromosomes of Trypanosoma brucei. In the Brazilian isolates, evidence for recombination was detected in 27 of the 36 chromosomes. Clustering analyses suggested two populations, in which two of the five older isolates from the 1990s clustered with a majority of recent isolates. Overall the analyses do not suggest individual sequence variants account for differences in clinical outcome or adaptation to different hosts. For the first known time, DNA of isolates from asymptomatic subjects were sequenced. Of interest, these displayed lower diversity than isolates from symptomatic subjects, an observation that deserves further investigation with additional isolates from asymptomatic subjects.

Technical note: Estimating body weight and body composition of beef cattle trough digital image analysis.

The use of digital images could be a faster and cheaper alternative technique to assess BW, HCW, and body composition of beef cattle. The objective of this study was to develop equations to predict body and carcass weight and body fat content of young bulls using digital images obtained through a Microsoft Kinect device. Thirty-five bulls with an initial BW of 383 (±5.38) kg (20 Black Angus, 390 [±7.48] kg initial BW, and 15 Nellore, 377 [±8.66] kg initial BW) were used. The Kinect sensor, installed on the top of a cattle chute, was used to take infrared light-based depth videos, recorded before the slaughter. For each animal, a quality control was made, running and pausing the video at the moment that the animal was standing with its body and head in line. One frame from recorded videos was selected and used to analyze the following body measurements: chest width, thorax width, abdomen width, body length, dorsal height, and dorsal area. From these body measurements, 23 indexes were generated and tested as potential predictors. The BW and HCW were assessed with a digital scale, whereas empty body fat (EBF) was estimated through ground samples of all tissues. To better understand the relationship among the measurements, the correlations between final BW (488 [±10.4] kg), HCW (287 [±12.5] kg), EBF (14 [±0.610] % empty BW) content, body measurements (taken through digital images), and developed indexes were evaluated. The REG procedure was used to develop the regressions, and the important independent variables were identified using the options STEPWISE and Mallow's Cp in the SELECTION statement. Chest width was the trait most related to weights and the correlations between this measurement and BW and HCW were above 0.85. The analysis of linear regressions between observed and predicted values showed that all models pass through the origin and have a slope of unity (null hypothesis [H]: = 0 and = 1; ≥ 0.993). The models to estimate BW and HCW of Angus and Nellore presented between 0.69 and 0.84 ( < 0.001), whereas from equations to estimate the EBF were lower ( = 0.43-0.45; ≤ 0.006). Index I5 [(chest width) × body length], related to the animal volume, was significant in all models created to estimate BW and HCW, and it explained more than 70% of the variation. This study indicates that digital images taken through a Microsoft Kinect system have the potential to be used as a tool to estimate body and carcass weight of beef cattle.

Sulfated galactofucan from Lobophora variegata: anticoagulant and anti-inflammatory properties.

Sulfated polysaccharides (fucans and fucoidans) from brown algae show several biological activities, including anticoagulant and anti-inflammatory activities. We have extracted a sulfated heterofucan from the brown seaweed Lobophora variegata by proteolytic digestion, followed by acetone fractionation, molecular sieving, and ion-exchange chromatography. Chemical analyses and 13C-NMR and IR spectroscopy showed that this fucoidan is composed of fucose, galactose, and sulfate at molar ratios of 1 : 3 : 2. We compared the anticoagulant activity of L. variegata fucoidan with those of a commercial sulfated polysaccharide (also named fucoidan) from Fucus vesiculosus and heparin. The experimental inflammation models utilized in this work revealed that fucoidan from L. variegata inhibits leukocyte migration to the inflammation site. Ear swelling caused by croton oil was also inhibited when sulfated polysaccharides from F. vesiculosus and L. variegata were used. The precise mechanism of different action between homo- and heterofucans is not clear; nevertheless, the polysaccharides studied here may have therapeutic potential in inflammatory disorders.

Genes at human chromosome 5q31.1 regulate delayed-type hypersensitivity responses associated with Leishmania chagasi infection.

Visceral leishmaniasis (VL) caused by Leishmania chagasi is endemic to northeast Brazil. A positive delayed-type hypersensitivity skin test response (DTH+) is a marker for acquired resistance to disease, clusters in families and may be genetically controlled. Twenty-three single nucleotide polymorphisms (SNPs) were genotyped in the cytokine 5q23.3-q31.1 region IRF1-IL5-IL13-IL4-IL9-LECT2-TGFBI in 102 families (323 DTH+; 190 DTH-; 123 VL individuals) from a VL endemic region in northeast Brazil. Data from 20 SNPs were analyzed for association with DTH+/- status and VL using family-based, stepwise conditional logistic regression analysis. Independent associations were observed between the DTH+ phenotype and markers in separate linkage disequilibrium blocks in LECT2 (OR 2.25; P=0.005; 95% CI=1.28-3.97) and TGFBI (OR 1.94; P=0.003; 95% CI=1.24-3.03). VL child/parent trios gave no evidence of association, but the DTH- phenotype was associated with SNP rs2070874 at IL4 (OR 3.14; P=0.006; 95% CI=1.38-7.14), and SNP rs30740 between LECT2 and TGFBI (OR 3.00; P=0.042; 95% CI=1.04-8.65). These results indicate several genes in the immune response gene cluster at 5q23.3-q31.1 influence outcomes of L. chagasi infection in this region of Brazil.

Atypical American visceral leishmaniasis caused by disseminated Leishmania amazonensis infection presenting with hepatitis and adenopathy.

Leishmania amazonensis is widely recognised as a cause of cutaneous leishmaniasis in Latin America, but it can also disseminate to produce atypical visceral leishmaniasis with hepatitis and lymphadenopathy. The patient, an 8-year-old Brazilian boy, presented with a febrile illness and hepatosplenomegaly, elevated liver enzymes and generalised adenopathy. Serological tests using antigens of L. chagasi, the typical cause of visceral leishmaniasis in Latin America, were inconclusive. Leishmania amazonensis was eventually isolated in a culture of a lymph node. The patient recovered fully after treatment with meglumine antimoniate. As this case illustrates, L. amazonensis produces a spectrum of disease that includes atypical American visceral leishmaniasis with evidence of hepatocellular injury and generalised lymphadenopathy.