Ancestral aneuploidy and stable chromosomal duplication resulting in differential genome structure and gene expression control in trypanosomatid parasites.

Aneuploidy is widely observed in both unicellular and multicellular eukaryotes, usually associated with adaptation to stress conditions. Chromosomal duplication stability is a tradeoff between the fitness cost of having unbalanced gene copies and the potential fitness gained from increased dosage of specific advantageous genes. Trypanosomatids, a family of protozoans that include species that cause neglected tropical diseases, are a relevant group to study aneuploidies. Their life cycle has several stressors that could select for different patterns of chromosomal duplications and/or losses, and their nearly universal use of polycistronic transcription increases their reliance on gene expansion/contraction, as well as post-transcriptional control as mechanisms for gene expression regulation. By evaluating the data from 866 isolates covering seven trypanosomatid genera, we have revealed that aneuploidy tolerance is an ancestral characteristic of trypanosomatids but has a reduced occurrence in a specific monophyletic clade that has undergone large genomic reorganization and chromosomal fusions. We have also identified an ancient chromosomal duplication that was maintained across these parasite's speciation, named collectively as the trypanosomatid ancestral supernumerary chromosome (TASC). TASC has most genes in the same coding strand, is expressed as a disomic chromosome (even having four copies), and has increased potential for functional variation, but it purges highly deleterious mutations more efficiently than other chromosomes. The evidence of stringent control over gene expression in this chromosome suggests that these parasites have adapted to mitigate the fitness cost associated with this ancient chromosomal duplication.

Control of Urochloa decumbens Using Glyphosate Applied by Remotely Piloted Aircraft and Ground Sprayer with Different Spray Nozzles.

The use of remotely piloted aircraft (RPA) to spray pesticides currently occurs, but knowledge about this technology is lacking due to the different locations, targets, and products applied. The objective of this study was to evaluate the control of Urochloa decumbens with glyphosate applied using an RPA (10 L ha-1) equipped with different spray nozzles (XR 11001 and AirMix 11001). For the purpose of comparison, ground application was also performed (100 L ha-1). The deposition was evaluated by means of the quantification of a tracer by spectrophotometry, the droplet spectrum was evaluated with water-sensitive paper, and the control efficiency was evaluated based on visual measurements with percentage scores. Statistical process control was used to analyse the quality of the deposition in the area. The results showed that the application via RPA presented a greater amount of tracer on the leaves than the ground application, suggesting that the former is a good option for application, even providing a lower coverage and number of droplets per area. Both application methods were effective at controlling Urochloa decumbens. The nozzles showed potential for use in applications, with control efficiency higher than 84% from 21 days after application. The percentage of droplets smaller than 100 µm in the applications was less than 5%. No nonrandom behaviour was observed during deposition, indicating a high-quality process.

Feline leishmaniasis in an animal shelter in northeastern Brazil: Clinical aspects, coinfections, molecular detection, and serological study of a new recombinant protein.

Infection and clinical cases of leishmaniasis caused by Leishmania infantum in cats have been increasingly reported in several countries, including Brazil. In this study, we used an enzyme-linked immunosorbent assay (ELISA) and an immunochromatographic test (ICT) based on a recombinant antigen (rKDDR-plus) to detect anti-Leishmania antibodies in cats from an animal shelter in northeastern Brazil. We compared the results with an ELISA using L. infantum crude antigen (ELISA-CA). We also investigated the presence of Leishmania DNA in blood or ocular conjunctival samples as well as the association between Leishmania PCR positivity and serological positivity to feline immunodeficiency virus (FIV), feline leukemia virus (FeLV) and Toxoplasma gondii. Concerning serological assays, a higher positivity was detected using the ICT-rKDDR-plus (7.5%; 7/93) as compared to ELISA-rKDDR-plus (5.4%; 5/93) and ELISA-CA (4.3%; 4/93). Upon PCR testing, 52.7% (49/93) of the ocular conjunctival swabs and 48.3% (44/91) of the blood samples were positive. Together, PCR and serological testing revealed overall positivities of 73.1% (68/93) and 12.9% (12/93), respectively. Among PCR-positive samples, 45.5% (31/68) showed co-infection with FIV, 17.6% (12/68) with FeLV, and 82.3% (56/68) with T. gondii. More than half of the PCR-positive cats showed at least one clinical sign suggestive of leishmaniasis (58.8%; 40/68) and dermatological signs were the most frequent ones (45.5%; 31/68). Both tests employing the recombinant antigen rKDDR-plus (i.e., ICT-rKDDR-plus and ELISA-rKDDR-plus) detected more positive cats than the ELISA-CA but presented low overall accuracy. PCR testing using either blood or ocular conjunctival samples detected much more positive cats than serological tests.

Long-term hematopoietic stem cells trigger quiescence in Leishmania parasites.

Addressing the challenges of quiescence and post-treatment relapse is of utmost importance in the microbiology field. This study shows that Leishmania infantum and L. donovani parasites rapidly enter into quiescence after an estimated 2-3 divisions in both human and mouse bone marrow stem cells. Interestingly, this behavior is not observed in macrophages, which are the primary host cells of the Leishmania parasite. Transcriptional comparison of the quiescent and non-quiescent metabolic states confirmed the overall decrease of gene expression as a hallmark of quiescence. Quiescent amastigotes display a reduced size and signs of a rapid evolutionary adaptation response with genetic alterations. Our study provides further evidence that this quiescent state significantly enhances resistance to treatment. Moreover, transitioning through quiescence is highly compatible with sand fly transmission and increases the potential of parasites to infect cells. Collectively, this work identified stem cells in the bone marrow as a niche where Leishmania quiescence occurs, with important implications for antiparasitic treatment and acquisition of virulence traits.

Indium Nitride Nanowires: Low Redox Potential Anodes for Lithium-Ion Batteries.

Advanced lithium-ion batteries (LIBs) are crucial to portable devices and electric vehicles. However, it is still challenging to further develop the current anodic materials such as graphite due to the intrinsic limited capacity and sluggish Li-ion diffusion. Indium nitride (InN), which is a new type of anodic material with low redox potential (<0.7 V vs Li/Li+) and narrow bandgap (0.69 eV), may serve as a new high-energy density anode material for LIBs. Here, the growth of 1D single crystalline InN nanowires is reported on Au-decorated carbon fibers (InN/Au-CFs) via chemical vapor deposition, possessing a high aspect ratio of 400. The binder-free Au-CFs with high conductivity can provide abundant sites and enhance binding force for the dense growth of InN nanowires, displaying shortened Li ion diffusion paths, high structural stability, and fast Li+ kinetics. The InN/Au-CFs can offer stable and high-rate Li delithiation/lithiation without Li deposition, and achieve a remarkable capacity of 632.5 mAh g-1 at 0.1 A g-1 after 450 cycles and 416 mAh g-1 at a high rate of 30 A g-1. The InN nanowires as battery anodes shall hold substantial promise for fulfilling superior long-term cycling performance and high-rate capability for advanced LIBs.

Instruments to assess diabetes knowledge, skills and attitudes of people living with diabetes mellitus: A COSMIN-based systematic review.

AIMS: Since knowledge, skills, and attitudes influence the adherence to self-care behaviours, the purpose of this review was to identify available instruments that measure diabetes knowledge, skills or attitudes of people living with this health condition and to evaluate their measurement properties. METHODS: A systematic search was conducted in PubMed, EMBASE, SCOPUS, Web of Science and CINAHL databases. The methodological quality of the included studies was assessed by the Consensus-based Standards for the selection of health Measurement Instruments. Risk of Bias checklist. The results of each study were rated by the updated criteria for good measurement properties and the quality of evidence was graded using a modified Grading of Recommendations Assessment, Development, and Evaluation approach. RESULTS: This review included 38 instruments that assess diabetes knowledge (n = 26) or skills (n = 6) or attitudes (n = 5) or both three constructs (n = 1). About 40% had satisfactory results for relevance, comprehensiveness, and comprehensibility, and about 30% had unsatisfactory results for comprehensiveness, but these results were supported by very low evidence. Most studies showed indeterminate results for structural validity (50%), internal consistency (98%) and reliability (84%). Instruments that had satisfactory results for construct validity and reliability were supported by low to very low evidence. One responsiveness study was identified, and the result was satisfactory, but supported by low evidence. The Diabetes Knowledge Questionnaire is the instrument with best evidence for use. CONCLUSIONS: To plan more assertive interventions, researchers need to follow up guidelines to develop instruments with good quality of the measurement proprieties.

Effects of in ovo injection of bacterial peptides and CpG-ODN on Salmonella enterica serovar Heidelberg infection in specific pathogen-free (SPF) chicks.

RESEARCH HIGHLIGHTS: Peptides + CpG-ODN reduced SH in caeca at the first week post-infection.Administered formulations did not reduce SH-faecal excretion.Levels of intestinal IgA were similar between all groups.CpG-ODN improved some parameters associated with chick intestinal health.

A Scoping Review of Pharmacists' and Pharmacy Students' Knowledge, Skills, and Attitudes in Medical Emergencies.

OBJECTIVES: Competence can be defined as a set of knowledge, skills, and attitudes. In a medical emergency scenario, competent pharmacists are increasingly required, mainly as a result of the expansion of professional functions in this context. Therefore, the objective of this study was to map the scientific evidence that shows the development of knowledge and/or skills, and/or attitudes in the training of pharmacists and pharmacy students to work in emergency care. FINDINGS: The scoping review was conducted in the PubMed, Embase, Latin American and Caribbean Health Sciences Literature, Scopus, Web of Science, and Cumulative Index to Nursing and Allied Health Literature databases in January 2021, as recommended by the Joanna Briggs Institute. Our study retrieved 6276 files, and 31 articles met the inclusion criteria. It was observed that the studies were developed mainly in the United States of America and addressed the development and/or assessment of knowledge and skills, and training in cardiac emergencies. The most used teaching strategy was simulation, and the most used assessment strategy was feedback and/or debriefing. SUMMARY: Publications involving the development of at least 1 domain of clinical competence have increased in the last decade. Thus, the mapping of studies has provided subsidies for identifying gaps in the teaching-learning process, as well as the identification of methodologies applied in the development and assessment of clinical competence for the referred population.

Ocular repercussions in COVID-19 patients: structural changes of the retina and choroid.

BACKGROUND: Neurotropic capabilities of SARS-COVs allow viruses to reach the central nervous system by hematogenous neuronal dissemination. The human retina, as an extension of the Central Nervous System, may have some neurodegenerative and/or vascular modifications related to COVID-19. OBJECTIVES: To evaluate choroidal and inner neural layers in participants previously recovered from COVID-19 compared to the control group using optical coherence tomography. METHODS: With a cross-sectional approach, the sample (n = 96), constituted by patients who have recovered from COVID-19 (n = 56) and healthy participants control group (n = 40) were ophthalmologically characterized. The neurodegenerative and vascular histological assessment was performed using SD-OCT and the mean thickness was measured in Early Treatment Diabetic Retinopathy Study (ETDRS) subfields. Retinal nerve fiber layer, Ganglion cell layer and subfoveal choroidal thickness were obtained through semi-automatic measurement. RESULTS: A total of 40 controls (27 women [67.5%]) and 56 COVID-19 participants (34 women [60.8%]) were included in this first report. There were retinal thickness significant differences in nearly all inner ETDRS subfields: nasal 3 mm (p = .025), I3 (p = .049), and temporal 3 mm (p = .009). Also, a decrease in neural layers was found in the nasal 3 mm (p = .049) and temporal 3 mm (p = .029) during ganglion cell layer assessment. The peripapillary retinal nerve fiber layer thickness was thinner in the COVID-19 group in superior temporal (p = .019), nasal (p = .002), inferior temporal (p = .046) and global (p = .014). Concerning the subfoveal choroidal measurement, an increase was observed in the COVID-19 group (p = .002). CONCLUSION: Participants who had recovered from COVID-19 showed a non-glaucomatous neuropathy trend pattern. We found differences closer to the classic description of the "bow-tie" observed in other neurological as compressive neuropathies at the chiasma location. OCT assessment also showed an increase in choroidal thickness as a result of vascular changes.

Milk and Fresh Cheese Quality of Crossbred Cows Supplemented with Phytogenic Additives and Managed under Thermal Stress.

This investigation aimed to assess the physiological parameters and quality of milk and fresh cheeses produced by cows that were housed in paddocks, either with or without shade, and supplemented with a phytogenic additive. Sixteen crossbred cows were allocated in a 4 × 4 Latin square design, dividing them into paddocks with or without shade, and providing or not providing a phytogenic additive in their feed. This resulted in a total of four treatment groups and sixteen experimental plots, each containing four animals, over four periods of 21 days. Various parameters were examined, including haematology, rectal and skin temperature, respiratory rate, milk yield and composition, serum parameters, and cheese yield and quality. It is worth noting that the temperature and humidity, as measured by a black globe thermometer, did not display significant variations between the different environments and exhibited minimal fluctuations throughout the day. Additionally, the supplementation of the phytogenic additive led to a reduction in haematocrit levels (p = 0.011). Furthermore, the analysis showed that whey obtained from cheese production had a higher fat content when cows were without access to shade (p = 0.005). Notably, there was an interaction between factors in relation to the total dry extract content, which was lower when cows had access to shade and received the additive (p = 0.010). In summary, the provision of a phytogenic additive and the presence or absence of shade did not bring about significant changes in milk production and quality or in the yield and quality of fresh cheese.

Ultrathin DSAEK versus DMEK - Review of systematic reviews.

The efficacy and safety of Descemet's membrane endothelial keratoplasty (DMEK) and ultrathin Descemet stripping automated endothelial keratoplasty (UT-DSAEK) have been recently compared in several systematic reviews (SRs). The aim of this study was to assess the evidence quality of such SRs, in order to obtain a scientifically rigorous comparison between the two techniques. We performed a systematic review of SRs and meta-analyses comparing the efficacy and safety between UT-DSAEK and DMEK up to 24th March 2023, using 3 electronic databases (PubMed, Cochrane Library, Google Scholar) plus manual reference search. Specific outcomes analyzed included best-corrected visual acuity (BCVA), endothelial cell density (ECD), rebubbling rate, and other postoperative complications. Of 90 titles/abstracts screened, four SRs met the inclusion criteria. All SRs adequately analyzed potential bias of the included studies. One SR raised concern for potential literature search bias and two SRs have heterogeneity in some outcomes analyzed. All SRs found higher BCVA after DMEK, but one SR reported significant heterogeneity. All SRs found significant heterogeneity in ECD analysis, with one SR providing inconsistent analysis of this outcome. Three SRs analyzed rebubbling rates, favoring UT-DSAEK over DMEK. Three SRs concluded a higher overall complication rate after DMEK, although rebubbling may be a confounding factor. This systematic review clarifies the strengths and weaknesses of published SRs and reinforces the conclusion that DMEK leads to superior visual outcomes compared to UT-DSAEK, with the trade-off of higher rebubbling rates and possibly other postoperative complications. Studies with longer follow-up are needed to ascertain these differences between procedures.

Simulated Drift of Dicamba and Glyphosate on Coffee Crop.

Weed management in areas adjacent to coffee plantations makes herbicide drift a constant concern, especially with the use of nonselective products such as dicamba. The objective of this study was to evaluate the phytotoxic effects of the herbicide dicamba alone and mixed with glyphosate as a result of simulated drift in a coffee-producing area. The study was conducted in duplicate at two different coffee cherry development stages. The study was performed with a randomized block design and a 2 × 5 + 1 factorial scheme with four replications using two herbicide spray solutions (dicamba and dicamba + glyphosate) and five low doses (0.25; 1; 5; 10; and 20%). Additionally, a control treatment without herbicide application was also employed. In this study, we evaluated the phytotoxic damage and biometric and productive parameters. Visual damages were observed with the use of dicamba and dicamba + glyphosate doses reduced by 0.25% to 5% in the first days after application. The main symptoms were new leaf epinasty, changes in the internodal distance, and plagiotropic branch curvature. Low doses led to reduced plant height and branch length. The treatments did not reduce productivity and performance but altered the physical classifications of grains.

Online information search by people with Multiple Sclerosis: A systematic review.

BACKGROUND: People with Multiple Sclerosis (pwMS) search for information online about various aspects of living with their disease, but details about patterns of searching and outcomes are unclear. This means that opportunities to leverage online resources to support pwMS, and to enhance shared decision making, may be missed. We aimed to do a systematic review of the literature on digital information searching by pwMS. METHODS: We performed a systematic search for studies assessing online information seeking of pwMS in MEDLINE and JSTOR databases. Studies were screened and selected by two investigators. All study designs were included, risk of bias was assessed using the Critical Appraisal Skills Programme qualitative checklist. Reports were assessed for the proportion of patients searching information online about MS, type of information sought, online tools used by patients, perceived quality of the information acquired, and impact of online searching in pwMS. RESULTS: We identified 5 studies, including 10,090 patients. Most pwMS search for information online (53.8-82 %), which they rarely discuss with physicians. The most common topics are treatment, general disease information, symptoms, lifestyle recommendations, prognosis, and coping strategies. Patients that are younger, have a shorter disease duration, primary progressive MS, and during periods of disease worsening, are more likely to use online resources. Online information is perceived as low quality by pwMS. CONCLUSIONS: Online information search is prevalent among pwMS. Despite concerns with the quality of the available information, only a minority of pwMS will discuss the information found with their physician. These findings highlight the importance of developing and providing quality online information resources for pwMS.

HIV pre-exposure prophylaxis and incidence of sexually transmitted infections in Brazil, 2018 to 2022: An ecological study of PrEP administration, syphilis, and socioeconomic indicators.

BACKGROUND: Human immunodeficiency virus (HIV) pre-exposure prophylaxis (PrEP) is one of the pillars of a combination prevention strategy for reducing the risk of new infections caused by HIV. The daily use of antiretroviral drugs by individuals who are not infected with HIV is required to prevent infection. Although its efficacy has been well established in the literature, in recent years, the decreased supply of antiretroviral drugs has been associated with an increase in the incidence of sexually transmitted infections (STI) and changes in the social determinants of health. An ecological study was conducted covering a five-year period (2018-2022), starting from the year of initiation of PrEP administration in Brazilian state capitals. PRINCIPAL FINDINGS: Descriptive analysis was performed, and the spatial distribution of study data was taken into account. Correlation analysis was used to assess the association between PrEP administration, the incidence and detection rate of STI, and socioeconomic data. The southern region showed the highest incidence rates of STI, but the northern and northeastern regions demonstrated the worst socioeconomic indicators, especially those related to illiteracy and basic sanitation. PrEP administration was significantly correlated with illiteracy (ρ = -0.658), per capita income (ρ = 0.622), public garbage collection (ρ = 0.612), syphilis (ρ = 0.628) and viral hepatitis (ρ = 0.419) incidences. Further, all STI were significantly associated with illiteracy and per capita income. SIGNIFICANCE: Our findings highlight the need to continue exploring PrEP use and rising syphilis rates. In terms of policy, PrEP administration appears to be inversely associated with regions of greater social vulnerability. Further efforts should focus on the social determinants and health needs of this population to improve access to PrEP and reduce social disparities.

3D-printed holder for drawing highly reproducible pencil-on-paper electrochemical devices.

Pencil drawing is one of the simplest and most cost-effective ways of fabricating miniaturized electrodes on a paper substrate. However, it is limited by the lack of reproducibility regarding the electrode drawing process. A 3D-printed pencil holder (3DPH) is proposed here for simple, reproducible, and low-cost hand-drawn fabrication of paper-based electrochemical devices. 3DPH was designed to keep pressure and angulation of the graphite mine constant on the paper substrate using a micromechanical pencil regardless of the user/operator. This approach significantly improved the reproducibility and cost of making reliable pencil-drawn electrodes. The results showed high reproducibility and accuracy of the 3DPH-assisted electrodes prepared by 4 different operators in terms of sheet resistance and electrochemical behavior. Cyclic voltammetric (CV) curves in the presence of [Fe(CN)6]3-/4- redox probe showed only 3.9% variation for the anodic peak currents of different electrodes prepared by different operators when compared with electrodes prepared without the 3D-printed support. SEM analyses revealed a more uniform graphite deposition/design of the electrodes prepared with 3DPH, which corroborates the results obtained by CV. As a proof of concept, 3DPH-assisted pencil-drawn graphite electrodes were employed for dopamine detection in synthetic saliva, showing a proportional increase in anodic peak current at 0.12 V vs. carbon pRE with increasing dopamine (DA) concentration, with a detection limit of 0.39µmol L-1. Moreover recovery was in the range 93-104% of DA (4-7% RSD) in synthetic saliva for three different concentrations, demonstrating the reliability of the approach. Finally, we believe this approach can make pencil-drawn technology more robust, accessible, reliable, and inexpensive for real on-site applications, especially in hard-to-reach locations or research centers with little investment.

3D hierarchical self-supporting Bi2Se3-based anode for high-performance lithium/sodium-ion batteries.

Bi2Se3 is a promising material for anodes in lithium-ion batteries (LIBs) and sodium-ion batteries (SIBs) due to its abundance, easy preparation, and high capacity. However, its practical application is hindered by low conductivity and significant volume variation during cycling, leading to poor rate capability and cycling stability. Herein, a novel composite consisting of Bi2Se3 nanoplates deposited on carbon cloth (CC) and encapsulated by reduced graphene oxide (rGO) has been designed and synthesized. The composite structure combines the advantages of the Bi2Se3 nanoplates, CC substrate, and rGO encapsulation, leading to enhanced electrochemical properties. The physical vapor deposition of Bi2Se3 nanoplates onto CC ensures a high loading of active material, while the rGO encapsulation provides a conductive and stable framework for the composite. This synergistic design allows for improved electron and ion transport, as well as efficient accommodation of the volume changes during cycling. In LIBs, the composite demonstrates a high reversible capacity of 467.5 mAh/g at 0.1 A/g after 120 cycles. Moreover, it displays an outstanding rate capability, delivering a capacity of 398.6 mAh/g at 5.0 A/g. Similarly, in SIBs, the composite maintains a reversible capacity of 375.3 mAh/g at 0.1 A/g over 100 cycles and exhibits a high-rate capacity of 286.3 mAh/g at 5.0 A/g. This work represents a significant step forward in addressing the challenges associated with Bi2Se3 as an anode material, paving the way for the development of high-performance LIBs and SIBs.

Emergence of zoonotic sporotrichosis due to Sporothrix brasiliensis in Minas Gerais, Brazil: A molecular approach to the current animal disease.

Sporotrichosis is a neglected fungal zoonosis with significant impacts on human and animal health. Accurate diagnosis, treatment, and understanding of the transmission dynamics of Sporothrix species are essential for mitigating the spread of sporotrichosis. This study aimed to identify the Sporothrix species involved in the ongoing outbreaks of animal sporotrichosis in the metropolitan region of Belo Horizonte, Minas Gerais, Brazil, and analyse the phylogenetic relationships between pathogenic species to investigate the outbreak origin. Additionally, to better understand the evolution of the disease, we conducted a retrospective survey of positive feline and canine cases from November 2017 to July 2021 with proven cultures for Sporothrix. A significant increase in animal cases over the last 4 years was observed, with cats being the most affected host. Sporothrix brasiliensis was the predominant agent in 100% of the clinical isolates (n = 180) molecularly identified. Phylogenetic and haplotype analysis points towards the cases isolated from Minas Gerais sharing the haplotype originating from a long-lasting outbreak of cat-transmitted sporotrichosis in Rio de Janeiro, however, with a secondary contribution from genotypes circulating in other outbreaks in Brazil. Thus, we present clear evidence of the circulation of different S. brasiliensis genotypes associated with animal sporotrichosis in the metropolitan region of Belo Horizonte. Genetic monitoring can contribute to understanding the causal agent for zoonotic sporotrichosis in epidemiological processes and help to implement disease prevention and control measures.

Complete assembly, annotation of virulence genes and CRISPR editing of the genome of Leishmania amazonensis PH8 strain.

We report the sequencing and assembly of the PH8 strain of Leishmania amazonensis one of the etiological agents of leishmaniasis. After combining data from long Pacbio reads, short Illumina reads and synteny with the Leishmania mexicana genome, the sequence of 34 chromosomes with 8317 annotated genes was generated. Multigene families encoding three virulence factors, A2, amastins and the GP63 metalloproteases, were identified and compared to their annotation in other Leishmania species. As they have been recently recognized as virulence factors essential for disease establishment and progression of the infection, we also identified 14 genes encoding proteins involved in parasite iron and heme metabolism and compared to genes from other Trypanosomatids. To follow these studies with a genetic approach to address the role of virulence factors, we tested two CRISPR-Cas9 protocols to generate L. amazonensis knockout cell lines, using the Miltefosine transporter gene as a proof of concept.

Complete Genome Sequences of Avian Metapneumovirus Subtype B Vaccine Strains from Brazil.

Avian metapneumovirus (aMPV) causes a highly contagious upper respiratory and reproductive disease in chickens, turkeys, and ducks. Here, complete genome sequences of aMPV-B vaccine strains BR/1890/E1/19 (PL21, Nemovac; Boehringer Ingelheim Animal Health, Brazil) and BR/1891/E2/19 (1062; Hipraviar, France) were sequenced and compared with the pathogenic field strain VCO3/60616.

Transcriptomic analysis of benznidazole-resistant and susceptible Trypanosoma cruzi populations.

BACKGROUND: Chagas disease (CD), caused by the parasite Trypanosoma cruzi, is a serious public health concern in Latin America. Nifurtimox and benznidazole (BZ), the only two drugs currently approved for the treatment of CD, have very low efficacies in the chronic phase of the disease and several toxic side effects. Trypanosoma cruzi strains that are naturally resistant to both drugs have been reported. We performed a comparative transcriptomic analysis of wild-type and BZ-resistant T. cruzi populations using high-throughput RNA sequencing to elucidate the metabolic pathways related to clinical drug resistance and identify promising molecular targets for the development of new drugs for treating CD. METHODS: All complementary DNA (cDNA) libraries were constructed from the epimastigote forms of each line, sequenced and analysed using the Prinseq and Trimmomatic tools for the quality analysis, STAR as the aligner for mapping the reads against the reference genome (T. cruzi Dm28c-2018), the Bioconductor package EdgeR for statistical analysis of differential expression and the Python-based library GOATools for the functional enrichment analysis. RESULTS: The analytical pipeline with an adjusted P-value of < 0.05 and fold-change > 1.5 identified 1819 transcripts that were differentially expressed (DE) between wild-type and BZ-resistant T. cruzi populations. Of these, 1522 (83.7%) presented functional annotations and 297 (16.2%) were assigned as hypothetical proteins. In total, 1067 transcripts were upregulated and 752 were downregulated in the BZ-resistant T. cruzi population. Functional enrichment analysis of the DE transcripts identified 10 and 111 functional categories enriched for the up- and downregulated transcripts, respectively. Through functional analysis we identified several biological processes potentially associated with the BZ-resistant phenotype: cellular amino acid metabolic processes, translation, proteolysis, protein phosphorylation, RNA modification, DNA repair, generation of precursor metabolites and energy, oxidation-reduction processes, protein folding, purine nucleotide metabolic processes and lipid biosynthetic processes. CONCLUSIONS: The transcriptomic profile of T. cruzi revealed a robust set of genes from different metabolic pathways associated with the BZ-resistant phenotype, proving that T. cruzi resistance mechanisms are multifactorial and complex. Biological processes associated with parasite drug resistance include antioxidant defenses and RNA processing. The identified transcripts, such as ascorbate peroxidase (APX) and iron superoxide dismutase (Fe-SOD), provide important information on the resistant phenotype. These DE transcripts can be further evaluated as molecular targets for new drugs against CD.