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Moringa oleifera is an Asian tree species cultivated for its nutritional and medicinal properties. Present study was carried out during 2021 and 2022 at Federal Institute of Minas Gerais, São João Evangelista-MG, Doce River Valley, South-Eastern Brazil to determine NPK doses for Moringa and critical NPK levels in the shoot. The experimental design was a fractional factorial of the type (1/2)43, with 32 treatments set up in four blocks. The treatments represented combinations of doses of N (0, 40, 80, 160 kg/ha); P2O5 (0, 45, 90, 180 kg/ha); and K2O (0, 20, 40, 80 kg/ha). Each experimental unit was composed of four seedlings arranged in a row. Plant shoot harvests were carried out every 60 days (January, March, May, July, September and November) for two consecutive years. The data were analyzed using regression analysis, and the adjusted response surface model was: Y = b0 + b1 N + b2 N2 + b3 P + b4 P2 + b5 K + b6 K2 + b7 NP + b8 NK + b9 PK. The productivity of fresh and dry matter of the shoot of M. oleifera plants was influenced only by N doses in the first year of cultivation, when the supply of 88.7 and 85.8 kg/ha of N led to the maximum estimated productivity for fresh (10,360.3 kg/ha) and dry matter (2,84.0 kg/ha). The critical contents in the shoot were 3.23 dag/kg (N), 0.41 dag/kg (P), and 2.18 dag/kg (K), while the annual extraction by plants was 78.44 kg/ ha (N), 8.99 kg/ha (P), and 48.81 kg/ha (K).

Over the years, tick-borne pathogens (TBPs) have garnered significant interest due to their medical, veterinary and economic importance. Additionally, TBPs have drawn attention to how these microorganisms interact with their own vectors, increasing the risk to human and animal infection of emerging and reemerging zoonoses. In this sense, ticks, which are obligate hematophagous ectoparasites, have a key role in maintaining and transmitting TBPs among humans and animals. The aim of this study was to assess the prevalence of neglected TBPs in mainland Portugal, namely Anaplasma spp., Babesia spp., Ehrlichia spp. and Neoehrlichia mikurensis. DNA fragments were detected in questing ticks collected from five different ecological areas under investigation. To the best of the authors’ knowledge, this study reports new worldwide findings, including B. bigemina infecting Ixodes frontalis, Ixodes ricinus and Rhipicephalus sanguineus sensu lato. Additionally, it presents new findings in Portugal of N. mikurensis infecting I. ricinus and of presumably Wolbachia endosymbionts being detected in I. ricinus. Overall, there were 208 tick samples that were negative for all screened TBPs. The results herein obtained raise concerns about the circulation of neglected TBPs in mainland Portugal, especially in anthropophilic ticks, highlighting the importance of adopting a One Health perspective.

Phytoremediation is a technique with high viability due to its low cost and minimal environmental impact. However, there are a few results regarding the use of tropical grasses in the phytoremediation of manganese (Mn), and there is no clear definition of which extractor is more suitable for assessing Mn availability in soils. The objective of this study was to evaluate the growth potential and phytoremediation of five tropical grasses in Mn-contaminated soil, as well as to assess the efficiency of four extractors in Mn extraction. The grass cultivars evaluated included Aruana and Tanzânia of Megathyrsus maximus, Xaraés and Marandu of Urochloa brizantha, and Basilisk of Urochloa decumbens grown in Typic Hapludox (Oxisol) amended with 0, 50, 150, and 450 mg of Mn per kg−1 of soil. The experiment was conducted in a greenhouse, using a completely randomized experimental design in a 5 × 4 factorial scheme, with five replications. The experimental period lasted for 90 days. The dry matter of the shoot and root of the grasses was evaluated. Mn concentration in dry matter and its availability in soil were extracted by Mehlich-1 and DTPA pH 7.3, while the semitotal concentration was determined by USEPA 3051 and total Mn by USEPA 3052 method. Tropical grasses showed reduced growth with increasing Mn rates in the soil, with the following sequence in the reduction in growth: Aruana > Basilisk > Marandu > Xaraés and lastly the Tanzânia. The Aruana cultivar showed greater potential for phytoextraction due to its higher ability to accumulate Mn in its biomass, followed by the Basilisk cultivar. The four different evaluated extractors were efficient in measuring Mn concentrations in the soil, with higher toxicity critical levels for the USEPA 3051 method due to the semitotal extraction nature and USEPA 3052 due to total Mn extraction. The growth of tropical grasses is reduced with increasing Mn rates applied to the soil. The Aruana of M. maximus grass has greater potential for phytoextraction of Mn-contaminated soils, followed by Basilisk of Urochloa decumbens, Xaraés and Marandu of U. brizantha, and Tanzânia of M. maximus. The Mehlich-1, DTPA pH 7.3, USEPA 3051, and 3052 extractors were efficient in extracting and evaluating Mn availability in soil for tropical grasses. The USEPA 3051 and 3052 methods presented higher toxicity critical levels.

Mountain insect biodiversity is unique, but is menaced by different drivers, particularly climate and land-use changes. In mainland Portugal, the highest mountain - Serra da Estrela - is one of the most important biodiversity hotspots, being classified as Natural Park since 1976. Many lepidopteran and odonate species, including rare and protected species, are known to occur in Serra da Estrela, but basic knowledge on their abundance, distribution and ecology is still lacking. Standardised sampling of these communities is crucial to provide valuable biological information to support short-term decision-making for conservation management, setting simultaneously the standards for mountain biodiversity monitoring aiming to tackle the effects of environmental change in the long-term.This study reports novel information on lepidopteran and odonate species diversity, distribution and abundance from Serra da Estrela Natural Park (Portugal). Seventy-two lepidopteran and 26 odonate species were sampled in this protected area, including the first findings of Apatura ilia (Denis & Schiffermüller, 1775), Macromia splendens (Pictet, 1843) and Vanessa virginiensis (Drury, 1773). New populations of Euphydrias aurinia (Rottemburg, 1775) and Oxygastra curtisii (Dale, 1834), protected species under the Habitats Directive, were found in this Natural Park and novel distribution and ecological data were collected for most species, including several rare species and subspecies [e.g. Aeshna juncea (Linnaeus, 1758), Coenonympha glycerion iphioides Staudinger, 1870, Cyaniris semiargus (Rottemburg, 1775) and Sympetrum flaveolum (Linnaeus, 1758)]. All data were collected using standardised sampling allowing its use as a baseline for biodiversity monitoring in Serra da Estrela.

Nickel (Ni) is extremely toxic to plants at high concentrations. Phytoliths have the potential to sequester the heavy metals absorbed by plants and act as a detoxification mechanism for the plant. The authors of the present study aimed to evaluate the effects of Ni on the growth and phytolith yield of grasses in two artificially contaminated soils. Two experiments separated by soil types (Typic Quartzipsamment and Rhodic Hapludox) were conducted in a completely randomized design in a 2 × 4 factorial scheme with three replications. The factors were two species of grass (Urochloa decumbens and Megathyrsus maximus) and three concentrations of Ni (20, 40, and 120 mg kg−1) and control treatment. The grasses were influenced by the increase in Ni rates in the soils. Ni exerted a micronutrient function with the addition of 30 mg kg−1 of Ni in soils, but this concentration caused toxicity in grasses. Such a level is lower than the limits imposed by the Brazilian environmental legislation. Higher Ni availability in Typic Quartzipsamment promoted Ni toxicity, with reduced growth and increased phytolith yield in the shoot, increased Ni in the shoot, and Ni occlusion in phytoliths by grasses, in comparison with Rhodic Hapludox. The yield and Ni capture in phytoliths by grasses in Ni-contaminated soils are related to the genetic and physiological differences between grasses and Ni availability in soils. Ni capture by phytoliths indicates that it may be one of the detoxification mechanisms of Urochloa decumbens to Ni contamination, providing additional tolerance. Megathyrsus maximus may be a future grass for the phytoremediation technique in Ni-contaminated soils.

The control of ticks through vaccination offers a sustainable alternative to the use of chemicals that cause contamination and the selection of resistant tick strains. However, only a limited number of anti-tick vaccines have reached commercial realization. In this sense, an antigen effective against different tick species is a desirable target for developing such vaccines. A peptide derived from the tick P0 protein (pP0) conjugated to a carrier protein has been demonstrated to be effective against the Rhipicephalus microplus, Rhipicephalus sanguineus, and Amblyomma mixtum tick species. The aim of this work was to assess the efficacy of this peptide when conjugated to the Bm86 protein against Dermacentor nitens and Ixodes ricinus ticks. An RNAi experiment using P0 dsRNA from I. ricinus showed a dramatic reduction in the feeding of injected female ticks on guinea pigs. In the follow-up vaccination experiments, rabbits were immunized with the pP0-Bm86 conjugate and challenged simultaneously with larvae, nymphs, and the adults of I. ricinus ticks. In the same way, horses were immunized with the pP0-Bm86 conjugate and challenged with D. nitens larva. The pP0-Bm86 conjugate showed efficacies of 63% and 55% against I. ricinus and D. nitens ticks, respectively. These results, combined with previous reports of efficacy for this conjugate, show the promising potential for its development as a broad-spectrum anti-tick vaccine.

Prior research found an association between mother–infant attachment and antibiotic use. Ambivalent-attached infants are more likely to take antibiotics than other infants, and their mothers tend to be less sensitive to their needs than most. This finding is important because it shows the association between psychological processes, early relationships, and health outcomes. We aim to learn about children with high-risk attachment relationships, such as disorganized-attached infants. This study compares antibiotic use, infant–mother interactive behavior, and health indicators according to infant attachment patterns (including disorganized attachment). For this purpose, we observed mothers–infants’ interactive behavior in free play at nine months and infants’ attachment in the Ainsworth Strange Situation at twelve months. Participants included 77 girls and 104 boys (full-term and preterm) and their mothers. Paradoxically, mothers of disorganized-attached infants reported that their children were ill only 1.56 times on average, but 61% of their children used antibiotics in the first nine months. The other mothers reported that their children were sick 5.73 times on average, but only 54% of their children used antibiotics in the same period. Infants with disorganized attachment had mothers who were more literate and less sensitive. These results add to a body of research that shows that early high-risk relationships affect children’s lives at multiple levels.

Abstract Background Mothers’ reports about pregnancy, maternity and their experiences during the perinatal period have been associated with infants’ later quality of attachment and development. Yet, there has been little research with mothers of very preterm newborns. This study aimed to explore mothers’ experiences related to pregnancy, premature birth, relationship with the newborn, and future perspectives, and to compare them in the context of distinct infants’ at-birth-risk conditions. Methods A semi-structured interview was conducted with women after birth, within the first 72 h of the newborn’s life. A total of 150 women participated and were divided in three groups: (1) 50 mothers of full-term newborns (Gestational Age (GA) ≥ 37 weeks; FT), (2) 50 mothers of preterm newborns (GA 32–36 weeks; PT) and (3) 50 mothers of very preterm newborns (GA

Mountain ecosystems are important biodiversity hotspots and valuable natural laboratories to study community assembly processes. Here, we analyze the diversity patterns of butterflies and odonates in a mountainous area of high conservation value—Serra da Estrela Natural Park (Portugal)—and we assess the drivers of community change for each of the two insect groups. The butterflies and odonates were sampled along 150 m transects near the margins of three mountain streams, at three elevation levels (500, 1000, and 1500 m). We found no significant differences in odonate species richness between elevations, but marginal differences (p = 0.058) were found for butterflies due to the lower number of species at high altitudes. Both insect groups showed significant differences in beta diversity (βtotal) between elevations, with species richness differences being the most important component for odonates (βrich = 55.2%), while species replacement drove the changes between butterfly assemblages (βrepl = 60.3%). Climatic factors, particularly those depicting harsher conditions of temperature and precipitation, were the best predictors of total beta diversity (βtotal) and its components (βrich, βrepl) for the two study groups. The study of insect biodiversity patterns in mountain ecosystems and of the role played by different predictors contribute to further our understanding on the community assembly processes and may help to better predict environmental change impacts on mountain biodiversity.

Anti-tick vaccines development mainly depends on the identification of suitable antigens, which ideally should have different features. These should be key molecules in tick biology, encoded by a single gene, expressed across life stages and tick tissues, capable of inducing B and T cells to promote an immunological response without allergenic, hemolytic, and toxic effects; and should not be homologous to the mammalian host. The discussion regarding this subject and the usefulness of “exposed” and “concealed” antigens was effectively explored in the publication by Nuttall et al. (2006). The present commentary intends to debate the relevance of such study in the field of tick immunological control.

Neoehrlichia mikurensis is a new emerging tick-borne Gram-negative bacterium, belonging to the family Anaplasmataceae, the main vector of which in Europe is the tick Ixodes ricinus. N. mikurensis is responsible for neoehrlichiosis, occurring mostly in patients with underlying diseases. In the present study, a total of 348 I. ricinus and Dermacentor reticulatus ticks collected in north-eastern Poland were analyzed for the prevalence of N. mikurensis. A total of 140 questing ticks (124 of I. ricinus ticks and 16 D. reticulatus) collected with the flagging method and 208 ticks (105 and 103 I. ricinus and D. reticulatus, respectively) removed from dogs were selected for the study. cDNA (questing ticks) and total DNA (questing and feeding ticks) were analyzed by qPCR targeting the 16S rRNA gene of N. mikurensis. Positive samples were further analyzed by nested PCR and sequencing. The prevalence differed between ticks collected from vegetation (19.3%; 27/140) and ticks removed from dogs (6.7%; 14/208). The presence of the pathogen in questing and feeding D. reticulatus ticks was proven in Poland for the first time. In summary, our research showed that infections of ticks of both the most common tick species I. ricinus and D. reticulatus in north-eastern Poland are present and ticks collected from urban areas were more often infected than ticks from suburban and natural areas. The detection of N. mikurensis in I. ricinus and D. reticulatus ticks from north-eastern Poland indicates potential transmission risk for tick-bitten humans at this latitude.

Antibiotics have individual and public-health drawbacks. Nevertheless, mother-infant attachment quality and maternal sensitivity are associated with antibiotic use. Ambivalent-attached infants are more likely to consume antibiotics than other infants. Conceivably, the emotional over-externalization of ambivalent-attached infants and maternal anxiety when infants are ill raise concerns in healthcare professionals, leading to antibiotic over-prescriptions. However, because infants prematurely born, particularly those with less than 32 weeks of gestation, are under more accurate health vigilance, the impact of infant and maternal behavior on antibiotic prescription may vanish in this sample. To test this hypothesis, we performed a longitudinal study to compare antibiotic use and the quality of mother-infant attachment in three groups: 86 infants born at full-term, 44 moderate-to-late preterm infants (32–36 gestation weeks), and 58 very-to-extreme preterm infants (

Rhipicephalus microplus is the only tick species known to serve as a biological vector of Theileria equi for horses and other equids in Brazil. The protozoan T. equi is one of the causal agents of equine piroplasmosis, a major threat in horse breeding systems. Vector competence is closely linked to the pathogens’ ability to evade tick defense mechanisms. However, knowledge of tick immune response against infections by hemoparasites of the Theileria genus is scarce. In the present study, the expression of genes involved in immune signaling pathways of R. microplus adults’ guts when challenged with a high or low parasitic load of T. equi was evaluated. This research demonstrates divergences in the immune gene expression pattern linked to T. equi infection in R. microplus since the Toll, IMD, and JNK signaling pathways were transcriptionally repressed in the guts of adult ticks infected with T. equi. Moreover, the results showed that different infectious doses of T. equi induce differential gene expression of key components of immune signaling cascades in R. microplus gut, suggesting a link between the intensity of infection and the activation of tick immunity response. The present study adds knowledge to elucidate the gut immune signaling response of R. microplus to T. equi infection. In addition, the generated data can serve as a basis for further investigations to develop strategies for controlling and preventing equine piroplasmosis.

Tick-borne rickettsioses (TBR) are caused by obligate, intracellular bacteria of the spotted-fever group (SFG) of the genus Rickettsia (Order Rickettsiales), transmitted by hard ticks. TBR are one of the oldest known vector-borne zoonoses and pose a threat to both human and animal health, as over the years, new SFG Rickettsia spp. have been reported worldwide with the potential to be human pathogens. In Portugal and Spain, the countries that constitute the Iberian Peninsula, reported TB rickettsiae causing human disease include Rickettsia conorii conorii, Rickettsia conorii israelensis, Rickettsia slovaca, Rickettsia raoultii, Candidatus Rickettsia rioja, Rickettsia sibirica mongolitimonae, and Rickettsia monacensis. An allochthonous case of TBR caused by Rickettsia massiliae, described in Spain, points to the need to monitor disease epidemiology, to predict risks of exposure and spread of disease, and taking into account globalization and climate changes. This review aims to provide up-to-date information on the status of TBR in the Iberian Peninsula, as well as to show the importance of a national and international collaborative epidemiology surveillance network, towards monitoring Rickettsia spp. circulation in both Portugal and Spain.

Rhipicephalus spp. (Acari: Ixodidae) ticks are obligate hematophagous arthropods, which constitute a model for the study of vector-host interactions. The chemical composition or elementome of salivary glands (SG) and cement provides information relevant for the study of protein-based complex multifunctional tissues with a key role in tick biology. In this study, we characterized the elementome of cement cones in Rhipicephalus sanguineus collected from naturally infested dogs and in SG and cement of R. bursa collected from experimentally infested rabbits at different feeding stages. The elementome was characterized using scanning electron microscopy (SEM) combined with energy dispersive X-ray spectroscopy (EDS). The results showed the identification of up to 14 chemical elements in the cement, and suggested tick/host-driven differences in the cement elementome between tick species and between SG and cement within the same species. By still unknown mechanisms, ticks may regulate cement elementome during feeding to affect various biological processes. Although these analyses are preliminary, the results suggested that N is a key component of the cement elementome with a likely origin in SG/salivary proteins (i.e., Glycine (C2H5NO2)-rich superfamily member proteins; GRPs) and other tick/host-derived components (i.e. NAPDH). Future research should be focused on tick elementome and its functional implications to better understand cement structure and function.

Bartonella spp. comprises emergent and re-emergent fastidious Gram-negative bacteria with worldwide distribution. Cats are the main reservoir hosts for Bartonella henselae and dogs represent opportunistic hosts for the bacteria. Even though ticks may also play a role in transmission, their competence as vectors for Bartonella spp. has not been totally understood. Considering only a few studies had a focus on screening Bartonella in animals, humans and ectoparasites in Portugal, this study aimed to address the molecular occurrence of Bartonella sp. in 123 stray cats, 25 stray dogs, 30 humans from Lisbon and 236 questing ticks within the country. Using a qPCR targeting the nuoG gene, it was possible to detect Bartonella sp. DNA on 20.32% of cat samples (25/123). From these positive samples, 13 sequences were characterized as B. henselae, 11 as B. clarridgeiae and 1 presented co-infection with both species. The absolute quantification of nuoGBartonella DNA in sampled cats ranged from 2.78 × 10 to 1.03 × 105 copies/µL. The sampled dogs, humans and ticks were negative. These results showed that B. henselae and B. clarridgeiae are circulating in stray cats from Lisbon. Additional and more extended studies should be conducted to determine the impact of such infections on humans, particularly those in constant and direct contact with cats.

The tick vector Rhipicephalus sanguineus is established as a complex of closely related species with high veterinary-medical significance, in which the presence of different genetic, morphological, and biological traits has resulted in the recognition of different lineages within taxa. One of the most striking differences in the “temperate” and “tropical” lineages of R. sanguineus (s.l.) is the vector competence to Ehrlichia canis, suggesting that these ticks tolerate and react differently to pathogen infection. The present study addresses the SG and MG proteome of the R. sanguineus tropical and temperate lineages and compares their proteomic profile during E. canis infection. Batches of nymphs from the two lineages were allowed to feed on naïve and experimentally E. canis infected dogs and after molting, adults were dissected, and salivary glands and midgut tissues separated. Samples were screened for the presence of E. canis before proteomic analyses. The representation of the proteins identified in infected and non-infected tissues of each lineage was compared and gene ontology used for protein classification. Results highlight important differences in those proteomic profiles that added to previous reported genetic, biological, behavioral, and morphological differences, strengthening the hypothesis of the existence of two different species. Comparing infected and non-infected tissues, the results show that, while in midgut tissues the response to E. canis infection is similar in the salivary glands, the two lineages show a different pattern of protein representation. Focusing on the proteins found only in the infected condition, the data suggests that the cement cone produced during tick feeding may be implicated in pathogen infection. This study adds useful information to the debate on the controversial R. sanguineus systematic status, to the discussion related with the different vectorial competence occurring between the two lineages and identifies potential targets for efficient tick and tick-borne disease control.

Fungisome® (F), a liposomal amphotericin B (AmB) product, is marketed in India as a safe and effective therapeutic for the parasitic infection visceral leishmaniasis. Its potential in the treatment of cutaneous leishmaniasis (CL), a disfiguring form of the disease affecting the skin, is currently unknown. Here, we report the evaluation of the efficacy of F in the Leishmania major BALB/c murine model of CL, including a head-to-head comparison with the standard liposomal AmB formulation AmBisome® (A). Upon intravenous administration at dose levels of 5, 10 and 15 mg/kg of body weight (on days 0, 2, 4, 6 and 8), F showed clear signs of toxicity (at 15 mg/kg), while A did not. After complete treatment (day 10), the tolerated doses of 5 and 10 mg/kg F had significant antileishmanial activity (ED50 = 4.0 and 12.8 mg/kg for qPCR-based parasite load and lesion size, respectively), although less than that of A at identical doses (ED50 = 3.0 and 8.8 mg/kg). The efficacy of F was inferior compared to A because lower levels of the active agent AmB accumulated within the infected lesion. In conclusion, despite possibly being less safe and efficacious than A at equivalent doses, the moderate in vivo activity of F could indicate a role in the systemic pharmacotherapy of CL. Keywords: Cutaneous leishmaniasis, Amphotericin B, Liposome, Efficacy, Pharmacokinetics

A system biology approach was used to gain insight into tick biology and interactions between vector and pathogen. Rhipicephalus annulatus is one of the main vectors of Babesia bigemina which has a massive impact on animal health. It is vital to obtain more information about this relationship, to better understand tick and pathogen biology, pathogen transmission dynamics, and new potential control approaches. In ticks, salivary glands (SGs) play a key role during pathogen infection and transmission. RNA sequencing obtained from uninfected and B. bigemina infected SGs obtained from fed female ticks resulted in 6823 and 6475 unigenes, respectively. From these, 360 unigenes were found to be differentially expressed (p < 0.05). Reversed phase liquid chromatography–mass spectrometry identified a total of 3679 tick proteins. Among them 406 were differently represented in response to Babesia infection. The omics data obtained suggested that Babesia infection lead to a reduction in the levels of mRNA and proteins (n = 237 transcripts, n = 212 proteins) when compared to uninfected controls. Integrated transcriptomics and proteomics datasets suggested a key role for stress response and apoptosis pathways in response to infection. Thus, six genes coding for GP80, death-associated protein kinase (DAPK-1), bax inhibitor-1 related (BI-1), heat shock protein (HSP), heat shock transcription factor (PHSTF), and queuine trna-ribosyltransferase (QtRibosyl) were selected and RNA interference (RNAi) performed. Gene silencing was obtained for all genes except phstf. Knockdown of gp80, dapk-1, and bi-1 led to a significant increase in Babesia infection levels while hsp and QtRibosyl knockdown resulted in a non-significant decrease of infection levels when compared to the respective controls. Gene knockdown did not affect tick survival, but engorged female weight and egg production were affected in the gp80, dapk-1, and QtRibosyl-silenced groups in comparison to controls. These results advanced our understanding of tick–Babesia molecular interactions, and suggested new tick antigens as putative targets for vaccination to control tick infestations and pathogen infection/transmission.

In the wake of the ‘omics’ explosion of data, reverse vaccinology approaches are being applied more readily as an alternative for the discovery of candidates for next generation diagnostics and vaccines. Promising protective antigens for the control of ticks and tick-borne diseases can be discovered by mining available omics data for immunogenic epitopes. The present study aims to explore the previously obtained Rhipicephalus bursa sialotranscriptome during both feeding and Babesia infection, to select antigenic targets that are either membrane-associated or a secreted protein, as well as unique to the ectoparasite and not present in the mammalian host. Further, they should be capable of stimulating T and B cells for a potential robust immune response, and be non-allergenic or toxic to the host. From the R. bursa transcriptome, 5706 and 3025 proteins were identified as belonging to the surfaceome and secretome, respectively. Following a reverse genetics immunoinformatics pipeline, nine preferred candidates, consisting of one transmembrane-related and eight secreted proteins, were identified. These candidates showed a higher predicted antigenicity than the Bm86 antigen, with no homology to mammalian hosts and exposed regions. Only four were functionally annotated and selected for further in silico analysis, which examined their protein structure, surface accessibility, flexibility, hydrophobicity, and putative linear B and T-cell epitopes. Regions with overlapping coincident epitopes groups (CEGs) were evaluated to select peptides that were further analyzed for their physicochemical characteristics, potential allergenicity, toxicity, solubility, and potential propensity for crystallization. Following these procedures, a set of three peptides from the three R. bursa proteins were selected. In silico results indicate that the designed epitopes could stimulate a protective and long-lasting immune response against those tick proteins, reflecting its potential as anti-tick vaccines. The immunogenicity of these peptides was evaluated in a pilot immunization study followed by tick feeding to evaluate its impact on tick behavior and pathogen transmission. Combining in silico methods with in vivo immunogenicity evaluation enabled the screening of vaccine candidates prior to expensive infestation studies on the definitive ovine host animals.

This study intends to characterize the sialotranscriptome profile of Rhipicephalus (Boophilus) microplus in response to Theileria equi and identify genes of interest with differential genomic expression, indicating relevant targets in the tick–protozoan interactions. The experimental design consisted of RNA sequencing from uninfected and T. equi-infected R. microplus salivary glands (SGs) to obtain transcriptomic profiles for characterization and comparison. A total of 288,952 transcripts were obtained from both tick profiles, 3456 transcripts (p < 0.05) differentially expressed in response to T. equi infection. The uninfected SGs’ registered 231,179 transcripts, of which 155,359 were annotated. The most transcribed sequences were female-specific histamine binding protein and lipocalins. Regarding the T. equi-infected SGs, from the 238,964 assembled transcripts, 163,564 were annotated. The most transcribed sequences were histone demethylase JARID1 and Y-box-binding protein. Five transcripts (cystatin, arginase, nuclear factor κB kinase inhibitor subunit β (IκB), IκB delta, lysosomal-trafficking regulator, and reeler protein) presented the gene ontology (GO) category “response to protozoan” and were exclusively displayed in the T. equi-infected profile. The transcriptome of T. equi was also analyzed, registering 4728 hits. The study’s genetic and molecular information would be of great value for future studies and biotechnological applications envisaging disease control.

The pandemic declared by the World Health Organization due to the SARS-CoV-2 virus (COVID-19) awakened us to a reality that most of us were previously unaware of—isolation, confinement and the massive use of information and communication technologies, as well as increased knowledge of the difficulties and limitations of their use. This article focuses on the rapid implementation of low-cost technologies, which allow us to answer a fundamental question: how can near real-time monitoring and follow-up of the elderly and their health conditions, as well as their homes, especially for those living in isolated and remote areas, be provided within their care and protect them from risky events? The system proposed here as a proof of concept uses low-cost devices for communication and data processing, supported by Long-Range (LoRa) technology and connection to The Things Network, incorporating various sensors, both personal and in the residence, allowing family members, neighbors and authorized entities, including security forces, to have access to the health condition of system users and the habitability of their homes, as well as their urgent needs, thus evidencing that it is possible, using low-cost systems, to implement sensor networks for monitoring the elderly using the LoRa gateway and other support infrastructures.

The negative impact of ticks and tick-borne diseases on animals and human health is driving research to discover novel targets affecting both vectors and pathogens. The salivary glands are involved in feeding and pathogen transmission, thus are considered as a compelling target to focus research. In this study, proteomics approach was used to characterize Rhipicephalus bursa sialoproteome in response to Babesia ovis infection and blood feeding. Two potential tick protective antigens were identified and its influence in tick biological parameters and pathogen infection was evaluated. Results demonstrate that the R. bursa sialoproteome is highly affected by feeding but infection is well tolerated by tick cells. The combination of both stimuli shifts the previous scenario and a more evident pathogen manipulation can be suggested. Knockdown of ub2n led to a significative increase of infection in tick salivary glands but a brusque decrease in the progeny, revealing its importance in the cellular response to pathogen infection, which is worth pursuing in future studies. Additionally, an impact in the recovery rate of adults (62%), the egg production efficiency (45.75%), and the hatching rate (88.57 %) was detected. Building knowledge on vector and/or pathogen interplay bridges the identification of protective antigens and the development of novel control strategies.

Ticks are among the most prevalent blood-feeding arthropods, and they act as vectors and reservoirs for numerous pathogens. Sialotranscriptomic characterizations of tick responses to blood feeding and pathogen infections can offer new insights into the molecular interplay occurring at the tick-host-pathogen interface. In the present study, we aimed to identify and characterize Rhipicephalus bursa salivary gland (SG) genes that were differentially expressed in response to blood feeding and Babesia ovis infection. Our experimental approach consisted of RNA sequencing of SG from three different tick samples, fed-infected, fed-uninfected, and unfed-uninfected, for characterization and inter-comparison. Overall, 7,272 expressed sequence tags (ESTs) were constructed from unfed-uninfected, 13,819 ESTs from fed-uninfected, and 15,292 ESTs from fed-infected ticks. Two catalogs of transcripts that were differentially expressed in response to blood feeding and B. ovis infection were produced. Four genes coding for a putative vitellogenin-3, lachesin, a glycine rich protein, and a secreted cement protein were selected for RNA interference functional studies. A reduction of 92, 65, and 51% was observed in vitellogenin-3, secreted cement, and lachesin mRNA levels in SG, respectively. The vitellogenin-3 knockdown led to increased tick mortality, with 77% of ticks dying post-infestation. The reduction of the secreted cement protein-mRNA levels resulted in 46% of ticks being incapable of correctly attaching to the host and significantly lower female weights post-feeding in comparison to the control group. The lachesin knockdown resulted in a 70% reduction of the levels associated with B. ovis infection in R. bursa SG and 70% mortality. These results improved our understanding of the role of tick SG genes in Babesia infection/proliferation and tick feeding. Moreover, lachesin, vitellogenin-3, and secreted cement proteins were validated as candidate protective antigens for the development of novel tick and tick-borne disease control measures.

Understanding host-pathogen-tick interactions remains a vitally important issue that might be better understood by basic research focused on each of the dyad interplays. Pathogens gain access to either the vector or host during tick feeding when ticks are confronted with strong hemostatic, inflammatory and immune responses. A prominent example of this is the Babesia spp.—tick—vertebrate host relationship. Babesia spp. are intraerythrocytic apicomplexan organisms spread worldwide, with a complex life cycle. The presence of transovarial transmission in almost all the Babesia species is the main difference between their life cycle and that of other piroplasmida. With more than 100 species described so far, Babesia are the second most commonly found blood parasite of mammals after trypanosomes. The prevalence of Babesia spp. infection is increasing worldwide and is currently classified as an emerging zoonosis. Babesia microti and Babesia divergens are the most frequent etiological agents associated with human babesiosis in North America and Europe, respectively. Although the Babesia-tick system has been extensively researched, the currently available prophylactic and control methods are not efficient, and chemotherapeutic treatment is limited. Studying the molecular changes induced by the presence of Babesia in the vector will not only elucidate the strategies used by the protozoa to overcome mechanical and immune barriers, but will also contribute toward the discovery of important tick molecules that have a role in vector capacity. This review provides an overview of the identified molecules involved in Babesia-tick interactions, with an emphasis on the fundamentally important ones for pathogen acquisition and transmission.

Abstract Background Ticks represent a significant health risk to animals and humans due to the variety of pathogens they can transmit during feeding. The traditional use of chemicals to control ticks has serious drawbacks, including the selection of acaricide-resistant ticks and environmental contamination with chemical residues. Vaccination with the tick midgut antigen BM86 was shown to be a good alternative for cattle tick control. However, results vary considerably between tick species and geographic location. Therefore, new antigens are required for the development of vaccines controlling both tick infestations and pathogen infection/transmission. Tick proteins involved in tick-pathogen interactions may provide good candidate protective antigens for these vaccines, but appropriate screening procedures are needed to select the best candidates. Methods In this study, we selected proteins involved in tick-Anaplasma (Subolesin and SILK) and tick-Babesia (TROSPA) interactions and used in vitro capillary feeding to characterize their potential as antigens for the control of cattle tick infestations and infection with Anaplasma marginale and Babesia bigemina. Purified rabbit polyclonal antibodies were generated against recombinant SUB, SILK and TROSPA and added to uninfected or infected bovine blood to capillary-feed female Rhipicephalus (Boophilus) microplus ticks. Tick weight, oviposition and pathogen DNA levels were determined in treated and control ticks. Results The specificity of purified rabbit polyclonal antibodies against tick recombinant proteins was confirmed by Western blot and against native proteins in tick cell lines and tick tissues using immunofluorescence. Capillary-fed ticks ingested antibodies added to the blood meal and the effect of these antibodies on tick weight and oviposition was shown. However, no effect was observed on pathogen DNA levels. Conclusions These results highlighted the advantages and some of the disadvantages of in vitro tick capillary feeding for the characterization of candidate tick protective antigens. While an effect on tick weight and oviposition was observed, the effect on pathogen levels was not evident probably due to high tick-to-tick variations among other factors. Nevertheless, these results together with previous results of RNA interference functional studies suggest that these proteins are good candidate vaccine antigens for the control of R. microplus infestations and infection with A. marginale and B. bigemina.

Ticks are obligate haematophagous ectoparasites of wild and domestic animals as well as humans, considered to be second worldwide to mosquitoes as vectors of human diseases. Tick-borne diseases are responsible worldwide for great economic losses in terms of mortality and morbidity of livestock animals. This review concerns to the different tick and tick-parasites control methods having a major focus on vaccines. Control of tick infestations has been mainly based on the use of acaricides, a control measure with serious drawbacks, as responsible for the contamination of milk and meat products, as a selective factor for acaricide-resistant ticks and as an environmental contaminant. Research on alternatives to the use of acaricides is strongly represented by tick vaccines considered a more cost-effective and environmentally safe strategy. Vaccines based on the Bm86 tick antigen were used in the first commercially available cattle tick vaccines and showed good results in reducing tick numbers, affecting weight and reproductive performance of female ticks which resulted in reduction of cattle tick populations over time and consequently lower reduction of the pathogen agents they carry.

Neste artigo apresentaremos uma breve perspectiva de um projecto de investigação que se centrou nacompetência de intercompreensã o, que, por sua vez, se encontra associada às competências decomunicação intercultural e plurilingual. A nossa intenção foi despertar nos nossos aprendentes acuriosidade para com outras línguas e culturas para além da portuguesa e inglesa. Escolhemos, assim, oAlemão e o Neerlandês, por pertencerem, tal como o Inglês, à mesma família linguística e, principalmente,por apresentarem enquanto línguas bastantes semelhanças. Quisemos sensibilizar os nossos aprendentespara as línguas germânicas, de forma a terem a oportunidade de alargarem os seus horizontes linguísticose culturais. Para tal elaborámos várias estratégias e materiais sempre com o objectivo de os mantermotivados e interessados em conhecer o desconhecido.Nesta breve reflexão, apresentaremos as competências fulcrais do nosso projecto, isto é, a competência deintercompreensão, a competência de comunicação intercultural e a competência plurilingues, assim comotambém as estratégias e os materiais usados em quatro aulas de sensibilização às três línguas germânicas.

Ticks are obligate haematophagous ectoparasites of wild and domestic animals as well as humans, considered to be second worldwide to mosquitoes as vectors of human diseases. Tick-borne diseases are responsible worldwide for great economic losses in terms of mortality and morbidity of livestock animals. This review concerns to the different tick and tick-parasites control methods having a major focus on vaccines. Control of tick infestations has been mainly based on the use of acaricides, a control measure with serious drawbacks, as responsible for the contamination of milk and meat products, as a selective factor for acaricide-resistant ticks and as an environmental contaminant. Research on alternatives to the use of acaricides is strongly represented by tick vaccines considered a more cost-effective and environmentally safe strategy. Vaccines based on the Bm86 tick antigen were used in the first commercially available cattle tick vaccines and showed good results in reducing tick numbers, affecting weight and reproductive performance of female ticks which resulted in reduction of cattle tick populations over time and consequently lower reduction of the pathogen agents they carry.

Phytoremediation is a technique with high viability due to its low cost and minimal environmental impact. However, there are a few results regarding the use of tropical grasses in the phytoremediation of manganese (Mn), and there is no clear definition of which extractor is more suitable for assessing Mn availability in soils. The objective of this study was to evaluate the growth potential and phytoremediation of five tropical grasses in Mn‐contaminated soil, as well as to assess the efficiency of four extractors in Mn extraction. The grass cultivars evaluated included Aruana and Tanzânia of Megathyrsus maximus, Xaraés and Marandu of Urochloa brizantha, and Basilisk of Urochloa decumbens grown in Typic Hapludox (Oxisol) amended with 0, 50, 150, and 450 mg of Mn per kg−1 of soil. The experiment was conducted in a greenhouse, using a completely randomized experimental design in a 5 × 4 factorial scheme, with five replications. The experimental period lasted for 90 days. The dry matter of the shoot and root of the grasses was evaluated. Mn concentration in dry matter and its availability in soil were extracted by Mehlich‐1 and DTPA pH 7.3, while the semitotal concentration was determined by USEPA 3051 and total Mn by USEPA 3052 method. Tropical grasses showed reduced growth with increasing Mn rates in the soil, with the following sequence in the reduction in growth: Aruana > Basilisk > Marandu > Xaraés and lastly the Tanzânia. The Aruana cultivar showed greater potential for phytoextraction due to its higher ability to accumulate Mn in its biomass, followed by the Basilisk cultivar. The four different evaluated extractors were efficient in measuring Mn concentrations in the soil, with higher toxicity critical levels for the USEPA 3051 method due to the semitotal extraction nature and USEPA 3052 due to total Mn extraction. The growth of tropical grasses is reduced with increasing Mn rates applied to the soil. The Aruana of M. maximus grass has greater potential for phytoextraction of Mn‐contaminated soils, followed by Basilisk of Urochloa decumbens, Xaraés and Marandu of U. brizantha, and Tanzânia of M. maximus. The Mehlich‐1, DTPA pH 7.3, USEPA 3051, and 3052 extractors were efficient in extracting and evaluating Mn availability in soil for tropical grasses. The USEPA 3051 and 3052 methods presented higher toxicity critical levels. [ABSTRACT FROM AUTHOR]

Nickel (Ni) is extremely toxic to plants at high concentrations. Phytoliths have the potential to sequester the heavy metals absorbed by plants and act as a detoxification mechanism for the plant. The authors of the present study aimed to evaluate the effects of Ni on the growth and phytolith yield of grasses in two artificially contaminated soils. Two experiments separated by soil types (Typic Quartzipsamment and Rhodic Hapludox) were conducted in a completely randomized design in a 2 × 4 factorial scheme with three replications. The factors were two species of grass (Urochloa decumbens and Megathyrsus maximus) and three concentrations of Ni (20, 40, and 120 mg kg−1) and control treatment. The grasses were influenced by the increase in Ni rates in the soils. Ni exerted a micronutrient function with the addition of 30 mg kg−1 of Ni in soils, but this concentration caused toxicity in grasses. Such a level is lower than the limits imposed by the Brazilian environmental legislation. Higher Ni availability in Typic Quartzipsamment promoted Ni toxicity, with reduced growth and increased phytolith yield in the shoot, increased Ni in the shoot, and Ni occlusion in phytoliths by grasses, in comparison with Rhodic Hapludox. The yield and Ni capture in phytoliths by grasses in Ni-contaminated soils are related to the genetic and physiological differences between grasses and Ni availability in soils. Ni capture by phytoliths indicates that it may be one of the detoxification mechanisms of Urochloa decumbens to Ni contamination, providing additional tolerance. Megathyrsus maximus may be a future grass for the phytoremediation technique in Ni-contaminated soils. [ABSTRACT FROM AUTHOR]