Distinct behavior of bovine-associated staphylococci species in their ability to resist phagocytosis and trigger respiratory burst activity by blood and milk polymorphonuclear leukocytes in dairy cows.

Mastitis affects a high proportion of dairy cows and is still one of the greatest challenges faced by the dairy industry. Staphylococcal bacteria remain the most important cause of mastitis worldwide. We investigated how distinct staphylococcal species evade some critical host defense mechanisms, which may dictate the establishment, severity, and persistence of infection and the outcome of possible therapeutic and prevention interventions. Thus, the present study investigated variations among distinct bovine-associated staphylococci in their capability to resist phagocytosis and to trigger respiratory burst activity of blood and milk polymorphonuclear neutrophil leukocytes (PMNL) in dairy cows. To do so, PMNL of 6 primiparous and 6 multiparous dairy cows were used. A collection of 38 non-aureus staphylococci (NAS) and 12 Staphylococcus aureus were included. The phagocytosis and intracellular reactive oxygen species (ROS) production by blood and milk PMNL were analyzed by flow cytometry. Phagocytosis, by both blood and milk PMNL, did not differ between S. aureus and NAS as a group, although within-NAS species differences were observed. Staphylococcus chromogenes (a so-called milk-adapted NAS species) better resisted phagocytosis by blood PMNL than the so-called environmental (i.e., Staphylococcus fleurettii) and opportunistic (i.e., Staphylococcus haemolyticus) NAS species. Otherwise, S. haemolyticus was better phagocytosed by blood PMNL than S. aureus, S. fleurettii, and S. chromogenes. No influence of the origin of the isolates within the staphylococci species in the resistance to phagocytosis by blood and milk PMNL was found. Overall, both S. aureus and NAS did not inhibit intracellular ROS production in blood and milk PMNL. Non-aureus staphylococci induced fewer ROS by milk PMNL than S. aureus, which was not true for blood PMNL, although species-specific differences in the intensity of ROS production were observed. Staphylococcus chromogenes induced more blood PMNL ROS than S. fleurettii and S. haemolyticus, and as much as S. aureus. Conversely, S. chromogenes induced fewer milk PMNL ROS than S. aureus. The origin of the isolates within the staphylococci species did not affect the ROS production by blood and milk PMNL. In conclusion, our study showed differences in staphylococci species in evading phagocytosis and triggering ROS production, which may explain the ability of some staphylococci species (i.e., S. aureus and S. chromogenes) to cause persistent infection and induce inflammation.

Integrated Proteomics Reveals Apoptosis-related Mechanisms Associated with Placental Malaria.

Malaria in pregnancy is a public health concern in malaria-endemic areas. Accumulation of maternal immune cells in the placenta and increased levels of inflammatory cytokines caused by sequestration of Plasmodium falciparum-infected erythrocytes have been associated to poor neonatal outcomes, including low birth weight because of fetal growth restriction. Little is known about the molecular changes occurring in a P. falciparum-infected placenta that has developed placental malaria during pregnancy but had the parasites cleared by pharmacological treatment (past infection). We conducted an integrated proteome, phosphoproteome and glycoproteome analysis in past P. falciparum-infected placentas aiming to find molecular changes associated with placental malaria. A total of 2946 proteins, 1733 N-linked glycosites and 4100 phosphosites were identified and quantified in this study, disclosing overrepresented processes related to oxidative stress, protein folding and regulation of apoptosis in past-infected placentas Moreover, AKT and ERK signaling pathways activation, together with clinical data, were further correlated to an increased apoptosis in past-infected placentas. This study showed apoptosis-related mechanisms associated with placental malaria that can be further explored as therapeutic target against adverse pregnancy outcomes.

Hormesis of 2,4-D choline salt in productive aspects of cotton.

The stimulating effect of a low dose of a substance considered to be toxic is known as hormesis. The aim of this work was to use dose-response curves to evaluate the hormesis effect provided by sub-doses of the herbicide 2,4-D choline salt on the productivity of cotton at different phenological stages. The experimental design was based on randomized blocks, with four repetitions and the treatments were distributed in a 9x3 factorial design, with nine fractions of the mean label dose of the herbicide 2,4-D choline salt formulation (0 (control); 0.4275; 0.855; 1.71; 3.42; 8.55; 17.1; 34.2 and 68.4 g a.e. ha-1) associated with three different phenological stage of cotton, namely: V4, B4 and C4. The plants were evaluated as to the main productive parameters of the cotton plant. When applied at the V4 stage, sub-doses of the herbicide 2,4-D choline salt negatively affect the cotton crop. Sub-doses between 0.82 and 2.23 g a.e. ha-1 of the herbicide 2,4-D choline salt applied at the B4 stage of cotton can increase all the productive variables of the crop. The productive aspects of cotton plants in the C4 stage were not influenced by the application of sub-doses of 2,4-D choline salt.

G6PD deficiency alleles in a malaria-endemic region in the Western Brazilian Amazon.

BACKGROUND: Plasmodium vivax parasites are the predominant cause of malaria infections in the Brazilian Amazon. Infected individuals are treated with primaquine, which can induce haemolytic anaemia in glucose-6-phosphate dehydrogenase (G6PD)-deficient individuals and may lead to severe and fatal complications. This X-linked disorder is distributed globally and is caused by allelic variants with a geographical distribution that closely reflects populations exposed historically to endemic malaria. In Brazil, few studies have reported the frequency of G6PD deficiency (G6PDd) present in malaria-endemic areas. This is particularly important, as G6PDd screening is not currently performed before primaquine treatment. The aim of this study was to determine the prevalence of G6PDd in the region of Alto do Juruá, in the Western Brazilian Amazon, an area characterized by a high prevalence of P. vivax infection. METHODS: Five-hundred and sixteen male volunteers were screened for G6PDd using the fluorescence spot test (Beutler test) and CareStart™ G6PD Biosensor system. Demographic and clinical-epidemiological data were acquired through an individual interview. To assess the genetic basis of G6PDd, 24 SNPs were genotyped using the Kompetitive Allele Specific PCR assay. RESULTS: Twenty-three (4.5%) individuals were G6PDd. No association was found between G6PDd and the number of malaria cases. An increased risk of reported haemolysis symptoms and blood transfusions was evident among the G6PDd individuals. Twenty-two individuals had the G6PDd A(-) variant and one the G6PD A(+) variant. The Mediterranean variant was not present. Apart from one polymorphism, almost all SNPs were monomorphic or with low frequencies (0-0.04%). No differences were detected among ethnic groups. CONCLUSIONS: The data indicates that ~1/23 males from the Alto do Juruá could be G6PD deficient and at risk of haemolytic anaemia if treated with primaquine. G6PD A(-) is the most frequent deficiency allele in this population. These results concur with reported G6PDd in other regions in Brazil. Routine G6PDd screening to personalize primaquine administration should be considered, particularly as complete treatment of patients with vivax malaria using chloroquine and primaquine, is crucial for malaria elimination.

Inflammasome activation and IL-1 signaling during placental malaria induce poor pregnancy outcomes.

Placental malaria (PM) is associated with severe inflammation leading to abortion, preterm delivery, and intrauterine growth restriction. Innate immunity responses play critical roles, but the mechanisms underlying placental immunopathology are still unclear. Here, we investigated the role of inflammasome activation in PM by scrutinizing human placenta samples from an endemic area and ablating inflammasome components in a PM mouse model. The reduction in birth weight in babies from infected mothers is paralleled by increased placental expression of AIM2 and NLRP3 inflammasomes. Using genetic dissection, we reveal that inflammasome activation pathways are involved in the production and detrimental action of interleukin-1ß (IL-1ß) in the infected placenta. The IL-1R pharmacological antagonist Anakinra improved pregnancy outcomes by restoring fetal growth and reducing resorption in an experimental model. These findings unveil that IL-1ß-mediated signaling is a determinant of PM pathogenesis, suggesting that IL-1R antagonists can improve clinical outcomes of malaria infection in pregnancy.

High prevalence of gestational night blindness and maternal anemia in a population-based survey of Brazilian Amazonian postpartum women.

Nutrition during pregnancy is one of the key elements to good maternal and child health, as well as to lifetime landmarks. However, many pregnant women go undernourished in less developed settings. The purpose of this study was to estimate the prevalence and factors associated with gestational night blindness (GXN) and maternal anemia in a cross-sectional population-based study in Cruzeiro do Sul, Acre State, Western Brazilian Amazon. All women living in the municipality admitted at the only maternity-hospital in the city to delivery of a singleton infant were eligible to this study (n = 1,525). Recruitment of participants took place between July 2015 to June 2016. GXN was assessed in the postpartum period by WHO standardized interview. Maternal anemia was defined as hemoglobin at delivery < 110.0 g/L. We estimated prevalence rates and adjusted prevalence ratios (aPR), alongside 95% confidence intervals (95% CI), of the factors associated with the outcomes through multiple Poisson regression models with robust variance. Alarming prevalence of GXN (11.5%; 95% CI, 9.97-13.25) and maternal anemia (39.4%; 95% CI, 36.84-41.95) were found. Factors associated with GXN were (aPR; 95% CI): ≥ 5 residents in the household (2.06; 1.24-3.41), smoking during pregnancy (1.78; 1.15-2.78), and attending < 6 antenatal care visits (1.61; 1.08-2.40). Factors associated with maternal anemia were (aPR; 95% CI): maternal age < 19 years (1.18; 1.01-1.38), gestational malaria (1.22; 1.01-1.49), not taking micronutrient supplements during pregnancy (1.27; 1.01-1.62), and attending < 6 antenatal care visits (1.40; 1.15-1.70). High prevalence rates of GXN and maternal anemia in these postpartum women may reflect poor assistance during antenatal care, underlying the importance of rethinking current protocols related to nutrition in pregnancy.

The Hidden Burden of Plasmodium vivax Malaria in Pregnancy in the Amazon: An Observational Study in Northwestern Brazil.

We measured the prevalence of malaria in pregnancy and estimated its impact on birth weight and length and maternal hemoglobin in 1,180 women from Juruá Valley, the main malaria hotspot in Brazil. Antenatal malaria episodes, 74.6% of them due to Plasmodium vivax, were microscopically diagnosed in 8.0% of the women and were associated with an average reduction in birth weight z-scores of 0.35 (95% confidence interval [CI] = 0.14-0.57) and in birth length z-scores of 0.31 (95% CI = 0.08-0.54), compared with malaria-free pregnancies. Affected mothers had a mean decrease in hemoglobin concentration at delivery of 0.33 g/100 mL (95% CI = 0.05-0.62 g/100 mL); 51.6% were anemic. The timing and frequency of antenatal infections influenced pregnancy outcomes and first- or second-trimester infections were not associated with decreased birth weight and length and maternal hemoglobin at delivery. Although repeated antenatal vivax infections were associated with poorer birth outcomes, even a single vivax malaria episode was associated with a significant reduction in birth weight and length and maternal hemoglobin. Overall, 7.5% women had the parasite's DNA found in peripheral blood at delivery. Most (83.1%) of these 89 perinatal infections were due to P. vivax and only 7.9% of them progressed to symptomatic disease after delivery. Plasmodium vivax and Plasmodium falciparum DNA was found in 0.6% and 0.3% of 637 cord blood samples examined, respectively, but only one newborn developed clinical neonatal malaria. Our results further challenge the notion that vivax malaria is relatively benign during pregnancy and call for better strategies for its prevention.

Global genetic diversity of var2csa in Plasmodium falciparum with implications for malaria in pregnancy and vaccine development.

Malaria infection during pregnancy, caused by the sequestering of Plasmodium falciparum parasites in the placenta, leads to high infant mortality and maternal morbidity. The parasite-placenta adherence mechanism is mediated by the VAR2CSA protein, a target for natural occurring immunity. Currently, vaccine development is based on its ID1-DBL2Xb domain however little is known about the global genetic diversity of the encoding var2csa gene, which could influence vaccine efficacy. In a comprehensive analysis of the var2csa gene in >2,000 P. falciparum field isolates across 23 countries, we found that var2csa is duplicated in high prevalence (>25%), African and Oceanian populations harbour a much higher diversity than other regions, and that insertions/deletions are abundant leading to an underestimation of the diversity of the locus. Further, ID1-DBL2Xb haplotypes associated with adverse birth outcomes are present globally, and African-specific haplotypes exist, which should be incorporated into vaccine design.

Malaria in Pregnancy Interacts with and Alters the Angiogenic Profiles of the Placenta.

Malaria in pregnancy remains a substantial public health problem in malaria-endemic areas with detrimental outcomes for both the mother and the foetus. The placental changes that lead to some of these detrimental outcomes have been studied, but the mechanisms that lead to these changes are still not fully elucidated. There is some indication that imbalances in cytokine cascades, complement activation and angiogenic dysregulation might be involved in the placental changes observed. Nevertheless, the majority of studies on malaria in pregnancy (MiP) have come from areas where malaria transmission is high and usually restricted to Plasmodium falciparum, the most pathogenic of the malaria parasite species. We conducted a cross-sectional study in Cruzeiro do Sul, Acre state, Brazil, an area of low transmission and where both P. vivax and P. falciparum circulate. We collected peripheral and placental blood and placental biopsies, at delivery from 137 primigravid women and measured levels of the angiogenic factors angiopoietin (Ang)-1, Ang-2, their receptor Tie-2, and several cytokines and chemokines. We measured 4 placental parameters (placental weight, syncytial knots, placental barrier thickness and mononuclear cells) and associated these with the levels of angiogenic factors and cytokines. In this study, MiP was not associated with severe outcomes. An increased ratio of peripheral Tie-2:Ang-1 was associated with the occurrence of MiP. Both Ang-1 and Ang-2 had similar magnitudes but inverse associations with placental barrier thickness. Malaria in pregnancy is an effect modifier of the association between Ang-1 and placental barrier thickness.

Hematological and biochemical characteristics during the transport of dog snapper Lutjanus jocu (Perciformes: Lutjanidae)

The objective of this study was to evaluate stress responses in dog snapper (Lutjanus jocu) during transport by evaluating their hematological and biochemical responses. Twenty-five wild dog snapper specimens were used in the experiment (220 ± 68 g and 24.5 ± 2.5 cm total length). Blood samples were collected prior to transport (control), and fish were placed in two transport boxes, one with anesthetic and one without anesthetic. Immediately after transport and after 24 h, blood was collected from the fish that underwent each treatment (with anesthetic and without anesthetic). Biochemical and hematological results demonstrated the inefficiency of benzocaine as a stress reliever during handling and transport. Biochemical parameters revealed the effects of stress during transport, and after 24 h, glucose levels and hematological parameters (hemoglobin, erythrocytes, leukocytes, neutrophils and MCH) showed a tendency to return to control levels. This study is the first to report stress response measurements of hematological and biochemical indicators in dog snapper, representing an important basis for the planning of future experiments involving the transport and handling of this fish species.(AU)

O objetivo desde estudo foi avaliar as repostas de estresse em dentão (Lutjanus jocu) durante o procedimento de transporte, através de respostas hematológicas e bioquímicas. Vinte e cinco exemplares selvagens de dentão foram utilizados no experimento (220 ± 68 g e 24.5 ± 2.5 cm de comprimento total). Foram realizadas coletas de sangue previamente ao transporte (controle), e os demais peixes foram acondicionados em duas caixas de transporte, uma com anestésico e outra sem anestésico. Imediatamente após o transporte e após 24 h, houve coleta de sangue para cada tratamento (com anestésico e sem anestésico). Os resultados bioquímicos e hematológicos apontam a ineficiência da benzocaína como mitigador do estresse durante o manuseio e transporte. Os parâmetros bioquímicos foram capazes de detectar o efeito do estresse durante o transporte, e após 24 h os níveis de glicose e alguns parâmetros hematológicos (hemoglobina, eritrócitos, leucócitos, neutrófilos e MCH) demonstraram uma tendência de retorno aos níveis do controle. Este trabalho é o primeiro a informar os níveis de resposta basal e de estresse para indicadores hematológicos e bioquímicos em dentão, representando uma base importante para o planejamento de futuras experiências com transporte e manejo dessa espécie.(AU)

Placental histopathological changes associated with Plasmodium vivax infection during pregnancy.

Histological evidence of Plasmodium in the placenta is indicative of placental malaria, a condition associated with severe outcomes for mother and child. Histological lesions found in placentas from Plasmodium-exposed women include syncytial knotting, syncytial rupture, thickening of the placental barrier, necrosis of villous tissue and intervillositis. These histological changes have been associated with P. falciparum infections, but little is known about the contribution of P. vivax to such changes. We conducted a cross-sectional study with pregnant women at delivery and assigned them to three groups according to their Plasmodium exposure during pregnancy: no Plasmodium exposure (n = 41), P. vivax exposure (n = 59) or P. falciparum exposure (n = 19). We evaluated their placentas for signs of Plasmodium and placental lesions using ten histological parameters: syncytial knotting, syncytial rupture, placental barrier thickness, villi necrosis, intervillous space area, intervillous leucocytes, intervillous mononucleates, intervillous polymorphonucleates, parasitized erythrocytes and hemozoin. Placentas from P. vivax-exposed women showed little evidence of Plasmodium or hemozoin but still exhibited more lesions than placentas from women not exposed to Plasmodium, especially when infections occurred twice or more during pregnancy. In the Brazilian state of Acre, where diagnosis and primary treatment are readily available and placental lesions occur in the absence of detected placental parasites, relying on the presence of Plasmodium in the placenta to evaluate Plasmodium-induced placental pathology is not feasible. Multivariate logistic analysis revealed that syncytial knotting (odds ratio [OR], 4.21, P = 0.045), placental barrier thickness (OR, 25.59, P = 0.021) and mononuclear cells (OR, 4.02, P = 0.046) were increased in placentas from P. vivax-exposed women when compared to women not exposed to Plasmodium during pregnancy. A vivax-score was developed using these three parameters (and not evidence of Plasmodium) that differentiates between placentas from P. vivax-exposed and unexposed women. This score illustrates the importance of adequate management of P. vivax malaria during pregnancy.

On the pathogenesis of Plasmodium vivax malaria: perspectives from the Brazilian field.

Life-threatening Plasmodium vivax malaria cases, while uncommon, have been reported since the early 20th century. Unfortunately, the pathogenesis of these severe vivax malaria cases is still poorly understood. In Brazil, the proportion of vivax malaria cases has been steadily increasing, as have the number of cases presenting serious clinical complications. The most frequent syndromes associated with severe vivax malaria in Brazil are severe anaemia and acute respiratory distress. Additionally, P. vivax infection may also result in complications associated with pregnancy. Here, we review the latest findings on severe vivax malaria in Brazil. We also discuss how the development of targeted field research infrastructure in Brazil is providing clinical and ex vivo experimental data that benefits local and international efforts to understand the pathogenesis of P. vivax.