Phylogenetic analysis of papillomaviruses in dogs from southern Brazil: molecular epidemiology and investigation of mixed infections and spillover events.

Papillomaviruses (PVs) have been identified in several animal species, including dogs (canine papillomaviruses, CPVs) and cattle (bovine papillomaviruses, BPVs). Although some BPVs may occasionally infect species other than cattle, to the best of our knowledge, BPVs have not been reported in dogs to date. Herein, we carried out a retrospective phylogenetic study of PVs circulating in dogs from southern Brazil between 2017 and 2022, also investigating possible mixed infections and spillover events. For this, we screened 32 canine papilloma samples by PCR using the degenerate primers FAP59/64 and/or MY09/11, which amplify different regions of the L1 gene; the genomic target often used for PV classification/typing. Out these, 23 PV DNA samples were successfully amplified and sequenced. All PVs amplified by FAP59/64 (n = 22) were classified as CPV-1. On the other hand, PVs amplified by MY09/11 (n = 4) were classified as putative BPV-1. Among these, three samples showed mixed infection by CPV-1 and putative BPV-1. One of the putative BPV-1 detected in co-infected samples had the L1 gene full-sequenced, confirming the gene identity. Furthermore, the phylogenetic classifications from the FAP59/64 and/or MY09/11 amplicons were supported by a careful in silico analysis, which demonstrated that the analysis based on them matches to the classification from the complete L1 gene. Overall, we described CPV-1 circulation in southern Brazil over the years and the potencial BPV infection in dogs (potential spillover event), as well as possible CPV/1/BPV-1 co-infections. Finally, we suggest the analysis of the complete genome of the putative BPVs detected in dogs in order to deepen the knowledge about the PV-host interactions.

Novel genomic targets for proper subtyping of bovine viral diarrhea virus 1 (BVDV-1) and BVDV-2.

Whole-genome phylogenetic analysis, the most suitable strategy for subtyping bovine viral diarrhea virus 1 (BVDV-1) and BVDV-2, is not feasible for many laboratories. Consequently, BVDV isolates/strains have been frequently subtyped based on analysis of single genomic regions, mainly the 5' untranslated region (UTR). This approach, however, may lead to inaccurate and/or poorly statistically supported viral classification. Herein, we describe novel primer sets whose amplicons may be easily sequenced and used for BVDV subtyping. Initially, genomic regions previously described as the most suitable targets for BVDV subtyping were analyzed for design of high-coverage primers. The putative amplicons were analyzed in silico for their suitability to reproduce the phylogenetic classification of 118 BVDV-1 and 88 BVDV-2 complete/near-complete genomes (CNCGs) (GenBank). This analysis was also performed considering the region amplifiable by primers HCV90-368, 324-326 and BP189-389 (5'UTR), which have been used for BVDV diagnosis and/or classification. After confirming the agreement between the analyses of our primers' amplicon versus the CNCGs, we optimized the RT-PCRs and evaluated their performance for amplification of BVDV isolates/strains (n = 35 for BVDV-1; n = 33 for BVDV-2). Among the potential targets for BVDV subtyping, we designed high-coverage primers for NS3-NS4A (BVDV-1) (526 bp amplicon) and NS5B (BVDV-2) (728 bp). The classification based on these regions fully reproduced the subtyping of all CNCGs. On the other hand, subtyping based on the putative amplicons from primers HCV90-368, 324-326 and BP189-389 showed disagreements in relation the CNCG analysis. The NS3-NS4A and NS5B primers also allowed the amplification of all BVDV isolates/strains tested. Finally, we suggest the use of these primers in future phylogenetic and epidemiological studies of BVDVs.

Seeing confusion through a new lens: on the impact of atoms of confusion on novices' code comprehension.

Code comprehension is crucial for software maintenance and evolution, but it can be hindered by tiny code snippets that can confuse the developers, called atoms of confusion. Previous studies investigated how atoms impact code comprehension through the perspectives of time, accuracy, and opinions of developers. However, we need more studies evaluating other perspectives and the combination of these perspectives on a common ground through experiments. In our study, we evaluate how the eye tracking method can be used to gain new insights when we compare programs obfuscated by the atoms with functionally equivalent clarified versions. We conduct a controlled experiment with 32 novices in Python and measure their time, number of attempts, and visual effort with eye tracking through fixation duration, fixations count, and regressions count. We also conduct interviews and investigate the subjects' difficulties with the programs. In our results, the clarified version of the code with Operator Precedence reduced the time spent in the region that contains the atom to the extent of 38.6%, and the number of answer attempts by 28%. Most subjects found the obfuscated version more difficult to solve than the clarified one, and they reported the order of precedence to be difficult to validate. By analyzing their visual effort, in the obfuscated version, we observed an increase of 47.3% in the horizontal regressions count in the atom region, making its reading more difficult. The additional atoms evaluated revealed other interesting nuances. Based on our findings, we encourage researchers to consider eye tracking combined with other perspectives to evaluate atoms of confusion and educators to favor patterns that do not impact the understanding and visual effort of undergraduates.

Phylogeny and amino acid analysis in single and mixed bovine papillomavirus infections in Southern Brazil, 2016-2020.

Bovine papillomaviruses (BPVs) exhibit a high degree of genetic variability, and several viral types have been identified based on analysis of the L1 gene. The L1 is the main capsid protein and the main target for neutralizing antibodies. We performed a retrospective study on BPVs circulating in Rio Grande do Sul state, Southern Brazil, in 2016-2020. DNA from 43 bovine papilloma samples were amplified using two degenerate primer sets - FAP59/64 and MY09/11 - targeting the L1 region, and analyzed for phylogeny, mixed BPV infections (coinfections) and amino acid (aa) sequences. We also performed an in silico analysis with 114 BPV L1 sequences from the GenBank database to assess the agreement between the phylogeny obtained based on complete L1 sequences versus that based on the region amplified using the FAP59/64 and MY09/11 primer sets. Considering single and coinfections, we identified 31 BPV-1 (31/43; 72.1%), 27 BPV-2 (27/43; 62.8%) and 4 BPV-6 (4/43; 9.3%). Coinfections with BPV-1 and BPV-2 were observed in 61.3% of the samples. Our results are supported by in silico analyses that demonstrate that the classification using FAP59/64 or MY09/11 matches the complete L1 results, except for BPV-17 and -18, which may be mistakenly classified depending on the primers used. Furthermore, we found unique or rare amino acids in at least one L1 sequence of each BPV type identified in our study, some of which have been identified previously in papillomavirus epitopes, suggesting immune-mediated selection. Finally, our study provides an overview of BPVs circulating in Southern Brazil over the last five years and point to the combined use of primers FAP59/64 and MY09/11 for analysis of BPV coinfections and putative epitopes.

A new (old) bovine viral diarrhea virus 2 subtype: BVDV-2e.

Bovine pestiviruses are members of the species Pestivirus A (bovine viral diarrhea virus 1, BVDV-1), Pestivirus B (BVDV-2) or Pestivirus H (HoBiPeV). To date, BVDV-2 isolates/strains have been classified into three subtypes (a-c) by phylogenetic analysis, and an additional subtype (d) has been proposed based on 5' untranslated region (UTR) secondary structures. In a previous study, we identified some BVDV-2 sequences in the GenBank database that could not be classified as subtype a, b or c by phylogenetic analysis of their genomes, UTRs or individual genes. Here, we performed a detailed study of these sequences and assessed whether they might represent a distinct BVDV-2 subtype. Initially, we collected 85 BVDV-2 complete/near-complete genomes (CNCGs) from GenBank and performed a "proof of equivalence" between phylogenetic analyses based on CNCGs and open reading frames (ORFs), which showed that ORFs may be reliably used as a reference target for BVDV-2 phylogeny, allowing us to increase our dataset to 139 sequences. Among these, we found seven sequences that could not be classified as BVDV-2a-c. The same was observed in the phylogenetic analysis of CNCGs and viral genes. In addition, the seven non-BVDV-2a-c sequences formed a distinct cluster in all phylogenetic trees, which we propose to term BVDV-2e. BVDV-2e also showed 44 amino acid changes compared to BVDV-2a-c, 20 of which are in well-defined positions. Importantly, an additional phylogenetic analysis including BVDV-2d and a pairwise comparison of BVDV-2e and BVDV-2d sequences also supported the difference between these subtypes. Finally, we propose the recognition of BVDV-2e as a distinct BVDV-2 subtype and encourage its inclusion in future phylogenetic analyses to understand its distribution and evolution.

Development, characterisation, stability study and antileukemic evaluation of nanoemulsions containing Astrocaryum aculeatum extract.

The objective of this work was to produce and characterise nanoemulsions containing tucumã extract and to evaluate the performance of the nanostructure and the free compound regarding antitumor activity, cytotoxicity, and oxidative metabolism in NB4/APL cells. The nanoemulsions showed adequate physicochemical characteristics (average size approx. 200 nm, polydispersity index less than 0.3, negative zeta potential and acid pH) maintained stable up to 90 days of storage in refrigeration condition. The nanoformulations did not present protein corona formation. Blank nanoemulsion treatments showed moderate toxicity. Furthermore, the nanoemulsion loaded with extract showed better antileukemic results than the free extract. However, nanoemulsions can be promising carriers of natural compounds, emphasising their biological properties and constituting alternatives in treating diseases.

Subtyping bovine viral diarrhea virus (BVDV): Which viral gene to choose?

Bovine viral diarrhea virus-1 (BVDV-1, Pestivirus A) and BVDV-2 (Pestivirus B) have been clustered into 21 and 4 subtypes, respectively. This genetic diversity, in addition to the lack of consensus on which genomic region to use for BVDV subtyping, has resulted in conflicting classifications depending on the target analyzed. Here, we investigated which genes or UTRs would reproduce the phylogeny obtained by complete genome (CG) analyses. The study was carried out with 91 (BVDV-1) and 85 (BVDV-2) CG available on GenBank database. The viruses were subtyped by analyzing their CG, as well as their individual genes and UTRs (complete 3' and 5'UTRs, and partial 5'UTR); and the phylogeny results were compared to each other. The sequences were aligned using the ClustalW multiple method (BioEdit Alignment Editor software, v.7.0.5.3) and the phylogenetic analyses were performed by the Maximum Likelihood method (MEGA-X software, v.10.2.4), with 1000 bootstrap replicates. The best analysis model for each gene/UTR was defined using the jModelTest software. The geodesic distance between the CG (reference) and individual genes/UTRs trees was also calculated (TreeCmp software, v.2.0). In general, 3'UTR-based analyses, followed by 5'UTR, presented the least reliable subtyping results. Regarding BVDV-1, phylogeny based on C, Erns, E1, E2, p7, NS2, NS3, NS4B, NS5A and NS5B was consistent with that of CG. In contrast, analyses performed with individual BVDV-2 genes showed at least one different clustering from the phylogeny based on the CG. After analyzing the geodesic distance between the CG and genes/UTRs trees, we observed that NS4B (for BVDV-1) and NS5A (BVDV-2) presented the closest topology and edge length to the CG analyses. Finally, comparing the phylogeny performed with the CG and the genes/UTRs, as well as the geodesic distance between them, we understand that NS4B and NS5A represent the most suitable targets for BVDV-1 and -2 subtyping, respectively, and may be considered in future phylogenetic studies.

Sequence analysis of the DA domain of glycoprotein E2 of pestiviruses isolated from beef cattle in Southern Brazil.

The envelope glycoprotein E2 of pestiviruses is a major target for neutralizing antibodies. In this study, we analyzed the E2 DA domain of 43 pestiviruses from Southern Brazil. The isolates were identified as Bovine viral diarrhea virus (BVDV) subtypes 1a and 1b or BVDV-2b. Compared to reference strains, the BVDV-1 and -2 isolates had four and two mutations in the DA domain, respectively. All BVDV-2 isolates had a deletion of residues 724 and 725. All mutated amino acids in the BVDV isolates had the same aa substitution, and all were in previously identified antibody binding sites. It is possible that an immunity-mediated selection is acting on the pestiviruses circulating in Southern Brazil.

Estimation of suspended sediment concentration in Guamá River in the Amazon region.

Monitoring of the suspended sediment concentration (SSC) is important for the analysis of morphological processes in watersheds and rivers, river habitats, and human activities associated with river management. Acoustic Doppler current profilers (ADCP) are used for measurements of flow and velocity in rivers through acoustic waves, which can also be converted into sediment concentrations. In this context, the objective is to verify the applicability of the use of ADCPs (Sontek YSI, model M9 River Surveyor) as an alternative for SSC estimates, this being a recent application of the equipment, mainly in rivers in the Amazon region rich in sediments in suspension. Thus, the Gartner method was used, calibrating its equation with the concentrations obtained by the point sampling method, collected with a bottle of Van Dorn in the Guamá River, Amazon, Brazil. The method used to obtain the SSC with data measured via ADCP resulted in a satisfactory estimate of the concentrations. The correlations between the measured and estimated sediment concentrations had R2 = 0.6827 for point sampling method and R2 = 0.8103 for vertically averaged SSC. These satisfactory results demonstrate the potential of using ADCP to estimate sediments suspended in Amazon basin.

Glycerol monolaurate nanocapsules for biomedical applications: in vitro toxicological studies.

The glycerol monolaurate (GML) is a surfactant used in the food industry and has potent antimicrobial activity against many microorganisms; however, the use of GML is not expanded due its high melting point and poor solubility in water. The aim of the study was to produce, characterize, and evaluate in vitro the cytotoxicity of GML and GML nanocapsules. The GML nanocapsules were produced and characterized by a mean diameter, zeta potential, and polydispersity index. The cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) release, thiobarbituric acid reactive substances (TBARS), and hemolytic activity. The genotoxicity was verified by comet assay. The physicochemical parameters showed a mean diameter of 192.5 ± 2.8 nm, a polydispersity index of 0.061 ± 0.018, and a zeta potential about - 21.9 ± 1 mV. The viability test demonstrated the protector effect of GML nanocapsule compared with the GML on peripheral blood mononuclear cells (PBMC) and VERO cells (isolated from kidney epithelial cells extracted from an African green monkey). A reduction in lipid peroxidation and lactate dehydrogenase release in GML nanocapsule-exposed cells compared with GML treated cells was observed. The damage on erythrocytes was addressed in treatment with GML, while the treatment with GML nanocapsules did not cause an effect. Moreover, the comet assay showed that the GML-caused genotoxicity and GML nanocapsules do not demonstrate damage. The study showed the reduction of toxicity of GML nanocapsules by many methods used in antimicrobial therapy.

New variants of canine parvovirus in dogs in southern Brazil.

Carnivore protoparvovirus 1 (canine parvovirus 2, CPV-2) has undergone a rapid evolution through mutations in the capsid protein VP2, giving rise to variants associated with unique clinicopathological and immunological features. VP2 is a major capsid protein involved in key steps of virus biology, including interactions with cellular receptors and with the immune system. This study analyzed the complete VP2 coding sequence of 38 CPV-2 isolates obtained from dogs with clinical parvovirosis in southern Brazil. Amplicons encompassing the whole VP2 coding region were subjected to nucleotide sequencing, and predicted amino acid sequences were analyzed to identify molecular markers of viral variants. Viral variants were classified as CPV-2a, -2b or -2c based on the presence of the amino acid Asn, Asp or Glu, respectively, at VP2 residue 426. Amino acid sequence analysis identified 20 CPV-2c and four CPV-2b isolates. Eleven viruses were identified as New CPV-2a, two as New CPV-2b, and one resembled the original CPV-2 and was designated CPV-2-like. In addition to the mutation at amino acid 426 of VP2, new 2a/2b variants containing a Ser297Ala mutation at residue 297 were identified. CPV-2-like samples contained some mutations that were also present in the original CPV-2 isolate, including as Leu, Thr, Ala and Asp at residues 87, 101, 300 and 305, respectively. The New CPV-2a isolates had three additional mutations (Phe267Tyr, Tyr324Ile and Thr440Ala) associated with selective pressure and development of disease in vaccinated dogs. The resemblance of the CPV-2-like isolate to CPV-2 suggests reemergence of CPV-2 and/or evolution from vaccine strains. Phylogenetic analysis grouped the variants with their respective reference strains, in general, according to amino acid changes. These results demonstrate the high VP2 diversity of CPV circulating in dogs in southern Brazil and indicate the emergence of new viral variants that differ markedly from the current vaccine strains.

Aesthetic Surgical Crown Lengthening Procedure.

The aim of this case report was to describe the surgical sequence of crown lengthening to apically reposition the dentogingival complex, in addition to an esthetic restorative procedure. Many different causes can be responsible for short clinical crown. In these cases, the correct execution of a restorative or prosthetic rehabilitation requires an increasing of the crown length. According to the 2003 American Academy of Periodontology (Practice Profile Survey), crown lengthening is the most habitual surgical periodontal treatment.

Influence of Pseudomonas putida AF7 inoculation on soil enzymes.

There has been some concern about the environmental impact of microbial agents. Pseudomonas may be used as bioremediator and as biopesticide. In this study, we report the use of soil enzyme assays as biological indicator of possible negative effects in soil functioning after the P. putida AF7 inoculation. For that, P. putida AF7 was originally isolated from the rizosphere of rice and was inoculated on three soil types: Rhodic Hapludox (RH), Typic Hapludox (TH); and Arenic Hapludult (AH). The acid phosphatase, beta-glucosidase and protease enzymes activities were measured for three period of evaluation (7, 14 and 21 days). In general, the enzymatic activities presented variation among the tested soils. The highest activities of beta-glucosidase and acid phosphatase were observed in the RH and AH soils, while the protease activity was higher in the TH soil. Also, the soil characteristics were measured for each plot. The activity of enzymes from the carbon cycle was positively correlated with the N and the P and the enzyme from the nitrogen cycle was negatively correlated with N and C.org. The presented data indicate that soil biochemical properties can be an useful tool for use as an indicator of soil perturbations by microbial inoculation in a risk assessment.

Qualidade das sementes de soja após a colheita com dois tipos de colhedora e dois períodos de armazenamento

Soybean mechanical harvest may bring qualitative losses to seeds. This study aimed to evaluate the quality of soybean seeds mechanically harvested by axial and conventional threshing systems, at different displacement speeds of the harvesters, before and after the storage period of six months. A randomized complete block design in a split-plot arrangement with four replications was used. The main plots were the harvest systems and the split-plots were the storage times. The harvest systems were: axial flow harvester, at 6, 8 and 10km h-1, conventional harvester, at 4 and 6km h-1, and manual harvest. Germination, percentage of normal seedlings, emergency speed index, sand emergency and mechanical damage of soybean seeds were evaluated. The results showed that the use of axial flow and conventional harvesters did not affect the emergency speed index, vigor and germination of soybean seeds. However, the axial flow system provided less mechanical damage. The increase of displacement speed, within the ranges of the operating parameters recommended by the manufacturer, did not change seed quality. Storage reduced seed vigor.

A colheita mecanizada de soja pode acarretar perdas qualitativas nas sementes. O presente trabalho teve como objetivo avaliar a qualidade de sementes de soja colhidas mecanicamente por sistemas axial e tangencial de trilha, em diferentes velocidades de avanço da colhedora, antes e após o período de armazenamento de seis meses. Utilizou-se o delineamento experimental de blocos casualizados, em esquema de parcela subdividida no tempo, com quatro repetições. Nas parcelas, foram avaliados os procedimentos de colheita e, nas subparcelas, as épocas de avaliação da qualidade da semente. Os procedimentos de colheita foram: colhedora com sistema de trilha axial, deslocando-se a 6, 8 e 10km h-1, colhedora com sistema de trilha tangencial (convencional), deslocando-se a 4 e 6km h-1, e colheita manual. Foram analisadas as seguintes variáveis: germinação, porcentagem de plântulas fortes, índice de velocidade de emergência (IVE), emergência em areia e injúria mecânica. Pôde-se concluir que o emprego das colhedoras com sistemas de trilha tangencial e axial não provocou diferenças no índice de velocidade de emergência, no vigor e na germinação das sementes de soja. No entanto, com relação à injúria mecânica, a colhedora axial mostrou-se superior à convencional. O incremento da velocidade de deslocamento, dentro dos parâmetros recomendados pelo fabricante, não alterou a qualidade das sementes. O armazenamento reduziu o vigor das sementes colhidas.

Qualidade das sementes de soja após a colheita com dois tipos de colhedora e dois períodos de armazenamento

Soybean mechanical harvest may bring qualitative losses to seeds. This study aimed to evaluate the quality of soybean seeds mechanically harvested by axial and conventional threshing systems, at different displacement speeds of the harvesters, before and after the storage period of six months. A randomized complete block design in a split-plot arrangement with four replications was used. The main plots were the harvest systems and the split-plots were the storage times. The harvest systems were: axial flow harvester, at 6, 8 and 10km h-1, conventional harvester, at 4 and 6km h-1, and manual harvest. Germination, percentage of normal seedlings, emergency speed index, sand emergency and mechanical damage of soybean seeds were evaluated. The results showed that the use of axial flow and conventional harvesters did not affect the emergency speed index, vigor and germination of soybean seeds. However, the axial flow system provided less mechanical damage. The increase of displacement speed, within the ranges of the operating parameters recommended by the manufacturer, did not change seed quality. Storage reduced seed vigor.

A colheita mecanizada de soja pode acarretar perdas qualitativas nas sementes. O presente trabalho teve como objetivo avaliar a qualidade de sementes de soja colhidas mecanicamente por sistemas axial e tangencial de trilha, em diferentes velocidades de avanço da colhedora, antes e após o período de armazenamento de seis meses. Utilizou-se o delineamento experimental de blocos casualizados, em esquema de parcela subdividida no tempo, com quatro repetições. Nas parcelas, foram avaliados os procedimentos de colheita e, nas subparcelas, as épocas de avaliação da qualidade da semente. Os procedimentos de colheita foram: colhedora com sistema de trilha axial, deslocando-se a 6, 8 e 10km h-1, colhedora com sistema de trilha tangencial (convencional), deslocando-se a 4 e 6km h-1, e colheita manual. Foram analisadas as seguintes variáveis: germinação, porcentagem de plântulas fortes, índice de velocidade de emergência (IVE), emergência em areia e injúria mecânica. Pôde-se concluir que o emprego das colhedoras com sistemas de trilha tangencial e axial não provocou diferenças no índice de velocidade de emergência, no vigor e na germinação das sementes de soja. No entanto, com relação à injúria mecânica, a colhedora axial mostrou-se superior à convencional. O incremento da velocidade de deslocamento, dentro dos parâmetros recomendados pelo fabricante, não alterou a qualidade das sementes. O armazenamento reduziu o vigor das sementes colhidas.

Qualidade das sementes de soja após a colheita com dois tipos de colhedora e dois períodos de armazenamento

Soybean mechanical harvest may bring qualitative losses to seeds. This study aimed to evaluate the quality of soybean seeds mechanically harvested by axial and conventional threshing systems, at different displacement speeds of the harvesters, before and after the storage period of six months. A randomized complete block design in a split-plot arrangement with four replications was used. The main plots were the harvest systems and the split-plots were the storage times. The harvest systems were: axial flow harvester, at 6, 8 and 10km h-1, conventional harvester, at 4 and 6km h-1, and manual harvest. Germination, percentage of normal seedlings, emergency speed index, sand emergency and mechanical damage of soybean seeds were evaluated. The results showed that the use of axial flow and conventional harvesters did not affect the emergency speed index, vigor and germination of soybean seeds. However, the axial flow system provided less mechanical damage. The increase of displacement speed, within the ranges of the operating parameters recommended by the manufacturer, did not change seed quality. Storage reduced seed vigor.

A colheita mecanizada de soja pode acarretar perdas qualitativas nas sementes. O presente trabalho teve como objetivo avaliar a qualidade de sementes de soja colhidas mecanicamente por sistemas axial e tangencial de trilha, em diferentes velocidades de avanço da colhedora, antes e após o período de armazenamento de seis meses. Utilizou-se o delineamento experimental de blocos casualizados, em esquema de parcela subdividida no tempo, com quatro repetições. Nas parcelas, foram avaliados os procedimentos de colheita e, nas subparcelas, as épocas de avaliação da qualidade da semente. Os procedimentos de colheita foram: colhedora com sistema de trilha axial, deslocando-se a 6, 8 e 10km h-1, colhedora com sistema de trilha tangencial (convencional), deslocando-se a 4 e 6km h-1, e colheita manual. Foram analisadas as seguintes variáveis: germinação, porcentagem de plântulas fortes, índice de velocidade de emergência (IVE), emergência em areia e injúria mecânica. Pôde-se concluir que o emprego das colhedoras com sistemas de trilha tangencial e axial não provocou diferenças no índice de velocidade de emergência, no vigor e na germinação das sementes de soja. No entanto, com relação à injúria mecânica, a colhedora axial mostrou-se superior à convencional. O incremento da velocidade de deslocamento, dentro dos parâmetros recomendados pelo fabricante, não alterou a qualidade das sementes. O armazenamento reduziu o vigor das sementes colhidas.