First report of root-knot nematode, Meloidogyne javaniva, infecting Stachys byzantina on São Paulo, Brazil.

Stachys byzantina belongs to the Labiatae and is known by the names "peixinho-da-horta" (Brazil) and "lamb's ear" (USA). Its importance is associated with its medicinal properties (Bahadori et al. 2020) and nutritional aspects (Milião et al. 2022). Root-knot nematodes cause severe damage to plants and suppress production. In January 2021, plants of S. byzantina in the municipality of Jaboticabal (21°14'38.7"S, 48°17'10.6"W) showed symptoms of reduced growth, yellowed leaves and the presence of galls in the roots. Initially, samples of roots from a S. byzantina were analyzed at the Nematology Laboratory (LabNema/UNESP), Jaboticabal, Brazil, estimating 20,000 eggs and juveniles of Meloidogyne sp. in 10 g of roots. To confirm the host ability of the species, a pathogenicity test was performed using Koch's postulate. For this purpose, the test was conducted in a greenhouse where 3,000 eggs and second-stage juveniles (J2) were inoculated onto three plants (n=3) of S. byzantina. After 90 days, the inoculated plants showed the same symptoms as those observed in the field. No symptom or nematode was detected in the uninoculated plant (control). Nematodes were extracted from the roots of inoculated plants and quantified. The perineal pattern of females (n=10) (Netscher and Taylor, 1974) and the labial region of males (n=10) (Eisenback and Hirschmann, 1981) were analyzed and compared with the morphological characteristics of the original description of the species (Chitwood, 1949). For analysis based on esterase isozyme phenotype, the α-method of Esbenshade and Triantaphyllou (1990) was used, and females (n=7) were examined. To confirm identification, whole genomic DNA from an adult female (n=1) was extracted using the Qiagen DNeasy® Blood & Tissue Kit and this sample was used for both genetic sequencing and the sequence-characterized amplified region techniques (SCAR). PCR amplifications were performed for the 18s rRNA gene using primers 988F and 1912R from Holterman et al (2006). Our sequence was deposited in GenBank (NCBI) under the identifier OP422209. Finally, species-specific SCAR primers (Fjav/Rjav, Me-F/Me-R, and Finc-F/Finc-R) designed by Zijlstra (2000) were used to identify Meloidogyne spp. Koch's postulate analysis yielded the following results: (n=1) 9,280 eggs and J2 (Reproduction factor, RF = 33.09); (n=2) 111,720 eggs and J2 (RF = 37.24); (n=3) 59,700 eggs and J2 (RF = 19.9) (RF mean = 30.08). The following characteristics were observed in the perineal region of females: Low and rounded trapezoidal dorsal arch with two distinct lateral lines clearly separating the dorsal and ventral arch regions, similar to the morphological features of the species description by Chitwood (1949). Males had a convex labial plate with a non-raised labial disk joining the submedial labia, a non-rugged labial region, the basal tubercles were usually wider than high, and a rounded tail tip (Eisenback and Hirschmann 1981). The α-esterase enzyme profile showed the J3 phenotype typical of M. javanica (Rm [×100] = 46.0, 54.5, and 58.9). The 18s rRNA sequences grouped Meloidogyne sp. with species such as M. enterolobii, M. incognita, and M. javanica. A DNA fragment of about 700 bp was amplified with Mj (Fjav/Rjav) primers, but not with Me (Me-F/Me-R) and Mi (Finc-F/Finc-R) primers, which confirmed the identification of M. javanica. Accurate identification and characterization of the occurrence of new hosts of M. javanica will allow us to determine the range and geographic distribution of the species. This is the first report on the occurrence of M. javanica on S. byzantina in Brazil. This report is important so that management strategies can be applied to prevent the spread of the pest to other areas.

First Report of Root-Knot Nematode Meloidogyne enterolobii Infecting Sweetpotato in the State of Rio Grande do Norte, Brazil.

The sweetpotato (Ipomoea batatas L., Convolvulaceae family) originated in Latin America and is currently cultivated worldwide. The storage roots, rich in calories, have made this crop one of the main caloric sources for low-income populations, especially in developing countries. Brazil annually produces about 805,000 tons, with the Northeast region responsible for 34% of this production (Albuquerque et al. 2020). In October 2019, sweetpotato plants cv. Campina, from a field in the region of Touros, state of Rio Grande do Norte (RN), Brazil (5°12'31"S 35°34'42"W), presented deformed storage roots, with galls, typical of root-knot nematodes. The roots were sent to the Nematology Laboratory (LabNema) where 14,032 eggs and 3,312 second-stage juveniles (J2s) of Meloidogyne sp., in 10 g of roots, were recovered. The species of adults was identified through morphological, biochemical, and phylogenetic analysis. The perineal region of females (n = 10) presented an oval shape, with a high and semi-trapezoidal dorsal arch and streak-free perivulval region. The labial region of males (n=10) presented high and rounded head cap, labial region slightly set off from the body, without annulations. The morphological characters were compatible with the original description of Meloidogyne enterolobii (Yang and Eisenback 1983). The phenotype of esterase isoenzymes showed two major bands (VS1-S1) also characteristic of M. enterolobii (Esbenshade and Triantaphyllou 1985). Sequences of 18S rDNA (~1200bp) of individual females (Holterman et al. 2006) obtained from sweetpotatoes before (SPme1 and 2) and after inoculation (SPme3 and 6), and from guava, used as M. enterolobii species control, were submitted to Bayesian analysis. The sequences presented genetic diversity among them resulting from seven SNPs (Single Nucleotide Polymorphism) and 99.4 to 99.9% identity with M. enterolobii sequences deposited in the NCBI GenBank (accession numbers MW209034-MW209039). The pathogenicity test was carried out under greenhouse conditions, in which 3,000 eggs and J2s from the original population isolated of M. enterolobii were inoculated in sweetpotato seedlings cv. Campina (n = 6). After three months, the roots presented galls and deformations typical of root-knot nematodes, while non-inoculated plants did not present any symptoms. An average of 15,900 eggs and J2s of M. enterolobii (RF = 5.3) were recovered from the roots, proving that sweetpotatoes were a host of this species. Meloidogyne enterolobii is known to cause great damage to sweetpotato (Ye et al. 2020). In Brazil, Meloidogyne nematode had been reported once, isolated from a sweetpotato field in the Ceara state and the species suggested by the authors according to esterase electrophoresis was M. enterolobii. Nonetheless, the authors did not present taxonomic, isoenzyme phenotypes and molecular species identification integratively, nor included pathogenicity tests (Silva et al. 2016). Therefore, it is the first time that M. enterolobii, with reliable identification by different methods, including sequencing, was detected in commercial sweetpotato fields in the RN state and in Brazil. The local farmers reported that this nematode deforms the storage roots which make them useless for commercialization, resulting in minimal losses of 50% of production in the infested areas. Furthermore, as sweetpotatoes are vegetatively propagated, the spread of this nematode through planting material is favored. Considering the importance of this crop in Brazil, this report is essential for control measures of this pathogen to be taken in order to avoid its spread to other regions.

Trueness and precision of the reproduction of dental casts by the conventional, milling and 3D printing techniques / Fidelidade e precisão de modelos de trabalho reproduzidos pelas técnicas convencional, de fresagem e de impressão 3D

Objective: To compare the reproduction trueness and precision of dental casts made by the conventional, milling and 3D printing techniques. Material and Methods: From an upper right side half-arch reference model (RM), 72 models were obtained and divided into three groups: conventional (CM), milled (MM) and printed (PM). All models were scanned and converted into standard tessellation language (.STL) files. The files were superimposed using 3D analysis software, and statistical analysis was performed using the root mean square (RMS) values obtained. The Shapiro-Wilk test was used to assess normality, and the Kruskal-Wallis test was used to compare groups (ρ < ⍺; ⍺ = 0.05). The Mann-Whitney U test was used for multiple comparisons among groups (ρ < ਕ; ਕ = 0.017). Results: There were significant differences in trueness (ρ = 0.000; ρ <0.001) and precision (ρ = 0.000; ρ < 0.001) among the three dental cast groups. Regarding trueness, CM presented better results, followed by MM and PM. Regarding precision, MM showed better results, followed by PM and CM, which did not show significant differences. Conclusions: For dental cast reproduction the conventional technique has the best trueness and the milling technique has the best precision.(AU)

Objetivo: Comparar a fidelidade e precisão de reprodução de modelos de trabalho pelas técnicas convencional, de fresagem e de impressão 3D. Material e Métodos: A partir de um modelo de referência (MR) de uma hemi-arcada superior direita, foram obtidos 72 modelos divididos em três grupos: convencionais (MC), fresados (MF) e impressos (MI). Todos foram digitalizados e convertidos sob a forma de ficheiros .STL (standard tesselation language). Os ficheiros foram sobrepostos utilizando um software de análise 3D, e através dos valores RMS (raiz do valor quadrático médio) obtidos foi realizada a análise estatística. Para avaliação da normalidade foi utilizado o teste Shapiro Wilk e para a comparação entre grupos foi utilizado o teste Kruskal-Wallis (ρ < ⍺; ⍺ = 0.05). Para as comparações múltiplas entre grupos, foi utilizado o teste U de Mann-Whitney (ρ < ਕ; ਕ = 0.017). Resultados:Existem diferenças significativas de fidelidade (ρ = 0.000; ρ <0.001) e precisão (ρ = 0.000; ρ < 0.001) entre os três grupos. Em relação à fidelidade, MC apresentou melhores resultados, seguido por MF e MI. Em relação à precisão, MF apresentou melhores resultados, seguido por MI e MC, que não apresentaram diferenças significativas entre si. Conclusão: Para reprodução de modelos de trabalho, a técnica convencional é a que apresenta maior fidelidade e a técnica de fresagem a que apresenta maior precisão.(AU)

First Report of Meloidogyne incognita Infecting Stachys byzantina in Brazil.

Stachys byzantina C. Koch (Lamiaceae alt. Labiatae), commonly known as lamb's ear, is an important medicinal plant with anti-inflammatory, antitumor, anticancer, antispasmodic, sedative and diuretic properties (Asnaashari et al. 2010). This plant is widely consumed in Europe and Asia as aromatic teas. In Brazil, it is an unconventional food plant, nonetheless, its medicinal properties have been recognized as well as its production. In May 2019, in a Sao Paulo State municipality, Jaboticabal, (21°14'38.7"S 48°17'10.6"W), S. byzantina plants presented reduced growth and chlorotic leaves associated with root galls. In the phytopathological clinic, 7,983 eggs and juveniles of Meloidogyne sp. were counted in 10 g of the plant roots. In 100 cm³ of soil surrounding the plant, 532 second-stage Meloidogyne sp. juveniles (J2) were found. Morphological, enzymatic and molecular identification of the nematode species found were performed (Fig. S1). For morphological analysis, perineal pattern of females (n = 10) and labial region of males (n = 10) were analyzed. In the perineal region of females, a high and trapezoidal dorsal arch with thick striations was observed, whereas the males presented the trapezoidal labial region with the prominent labial disc in relation to the sub-median lips and transverse streaks in the head region, typical characteristics of M. incognita (Kofoid and White, 1919) Chitwood 1949. (Netscher and Taylor 1974; Eisenback and Hirschmann 1981). The esterase enzyme profile, obtained individually from 8 females, was compatible with phenotype I1 [Rm (x100) = 46.25], also associated with M. incognita (Esbenshade and Triantaphyllou 1985). Molecular analysis was realized (n = 3) by applying the primers Finc/Rinc (Zijlstra et al. 2000) in the DNA of individual females, which resulted in the amplification of an amplicon of 1200 bp specific for M. incognita. Pathogenicity testing was conducted in a greenhouse by inoculation of 5,000 eggs and juveniles from the original population into S. byzantina seedlings (n = 4). After 90 days, the inoculated plants, unlike the non-inoculated ones, exhibited symptoms similar to those initially observed in the field. The nematodes were extracted from the roots of the inoculated plants, quantified, and the identity of M. incognita was confirmed. The average reproductive factor obtained was 136.6, confirming the pathogenicity of M. incognita to S. byzantina. Thus, this is the first report of M. incognita associated with S. byzantina in Brazil and in the world. Lamb's ear is a horticultural plant, and its high reproductive factor to M. incognita can also result in damage to the subsequent crops. In addition, Lamb's ear is propagated vegetatively and this favors the spread of nematodes to other areas. This new report is important in order to alert producers to realize the proper management of this nematode in S. byzantina.

Screening of Beneficial Microorganisms to Improve Soybean Growth and Yield

Abstract The objective of this research was to identify the best microorganisms, alone or in mixture for total biomass gain (root + shoot), positive change in gas exchange, nutrient uptake (root, shoot and grain) and yield and yield components in the soybean crop. Trial under greenhouse conditions had the experimental design in a completely randomized scheme with 26 treatments and four replicates. The treatments consisted of the rhizobacteria BRM 32109, BRM 32110 and 1301 (Bacillus sp.), BRM 32111 and BRM 32112 (Pseudomonas sp.), BRM 32113 (Burkholderia sp.), BRM 32114 (Serratia sp.), Ab-V5 (Azospirillum brasilense) and 1381 (Azospirillum sp.), and the fungus Trichoderma asperellum (a mix of the isolates UFRA.T06, UFRA.T09, UFRA.T12 and UFRA.T52). Besides, the same isolates were combined in pairs, completing 16 combinations. Control treatments received no microorganism. Microorganisms applied isolated or in combination, provided biomass gain, positive gas exchange, increases in nutrients uptake at the shoot and grain, and improved grain yield and its components than control plants. Stood out the combination Ab-V5 + T. asperellum pool, which provided a 25% improvement in grain yield.

Cognition and renal function: findings from a Brazilian population / Cognição e função renal: achados de uma população brasileira

ABSTRACT Introduction: The prevalence of chronic kidney disease (CKD) is increasing, with a potential impact in the risk of acceleration of dementia. The potential association between glomerular filtration rate (eGFR) and cognitive performance was scarcely studied. The aim of this study was to evaluate cognitive performance levels across different degrees of kidney function. Methods: We analyzed 240 outpatients in a nephrology service, classified according to eGFR: Advanced (≤ 30ml/min/1.73m2), Moderate (30,1ml/min/1.73m2 to ≤ 60ml/min/1.73m2), and Mild CKD (> 60ml/min/1.73m2). Word list memory, Semantic fluency, Mental State Mini Exam and Trail Making Test (TMT) were applied to evaluate cognitive performance. In the TMT, lower scores are associated with better cognition. In linear regression, cognitive function was considered as dependent variables while groups based on eGFR were considered explanatory variables. The group with eGFR > 60ml/min was the reference and models were adjusted for confounding factors. Results: In our population (n = 240) 64 patients (26.7%) were classified as having advanced, 98(40,8%) moderate, and 78(32,5%) mild. There was no statistical difference among them in MMSE or in the verbal fluency test. However, comparing to mild, patients with advanced CKD presented significantly worse cognitive performance measured by TMTA [50,8s ± 31.1s versus 66,6s ± 35,7s (p = 0.016)] and TMTB [92,7s ± 46,2s versus 162,4s ± 35,7s (p < 0.001)]. Significantly lower TMTB scores (CI95%) 33,0s (4,5-61,6s) were observed in patients with mild compared to advanced CKD in the multivariate analysis adjusting for age, education, sex, diabetes, and alcohol use. Conclusion: Advanced CKD is independently associated with poorer cognitive performance measured by an executive performance test compared to mild CKD.

RESUMO Introdução: A elevação da prevalência de doença renal crônica (DRC) traz consigo um impacto potencial sobre o risco de aceleração da demência. A possível associação entre taxa de filtração glomerular (TFGe) e desempenho cognitivo foi pouco estudada. O objetivo do presente estudo foi avaliar os níveis de desempenho cognitivo em indivíduos com diferentes graus de função renal. Métodos: Foram analisados 240 pacientes ambulatoriais atendidos em um serviço de nefrologia classificados segundo a TFGe em grupos com DRC avançada (≤ 30ml/min/1,73m2), moderada (30,1ml/min/1,73m2 a ≤ 60ml/min/1,73m2) ou leve (> 60ml/min/1,73m2). Testes de memória por listas de palavras, fluência semântica, o mini exame do estado mental e o teste das trilhas (TT) foram aplicados para avaliar o desempenho cognitivo. No TT, escores mais baixos representam melhor cognição. Na regressão linear, função cognitiva foi considerada como variável dependente, enquanto os grupos baseados na TFGe foram considerados como variáveis explicativas. O grupo com TFGe > 60ml/min foi utilizado como referência e os modelos foram ajustados para fatores de confusão. Resultados: Em nossa população (n = 240), 64 pacientes (26,7%) foram diagnosticados com DRC avançada, 98 (40,8%) com DRC moderada e 78 (32,5%) com DRC leve. Não houve diferença estatística entre eles no MEEM ou no teste de fluência verbal. Contudo, em relação aos indivíduos com DRC leve, os pacientes com DRC avançada apresentaram desempenho cognitivo significativamente pior medido pelo TT A [50,8s ± 31,1s x 66,6s ± 35,7s (p = 0,016)] e TT B [92,7s ± 46,2s x 162,4s ± 35,7s (p < 0,001)]. Escores significativamente mais baixos no TT B (IC95%) 33,0s (4,5-61,6s) foram observados nos pacientes com DRC leve em comparação com o grupo com DRC avançada na análise multivariada ajustada para idade, escolaridade, sexo, diabetes e uso de álcool. Conclusão: DRC avançada esteve independentemente associada a pior desempenho cognitivo medido por um teste de desempenho executivo em comparação à DRC leve.

Cognition and renal function: findings from a Brazilian population.

INTRODUCTION: The prevalence of chronic kidney disease (CKD) is increasing, with a potential impact in the risk of acceleration of dementia. The potential association between glomerular filtration rate (eGFR) and cognitive performance was scarcely studied. The aim of this study was to evaluate cognitive performance levels across different degrees of kidney function. METHODS: We analyzed 240 outpatients in a nephrology service, classified according to eGFR: Advanced (≤ 30ml/min/1.73m2), Moderate (30,1ml/min/1.73m2 to ≤ 60ml/min/1.73m2), and Mild CKD (> 60ml/min/1.73m2). Word list memory, Semantic fluency, Mental State Mini Exam and Trail Making Test (TMT) were applied to evaluate cognitive performance. In the TMT, lower scores are associated with better cognition. In linear regression, cognitive function was considered as dependent variables while groups based on eGFR were considered explanatory variables. The group with eGFR > 60ml/min was the reference and models were adjusted for confounding factors. RESULTS: In our population (n = 240) 64 patients (26.7%) were classified as having advanced, 98(40,8%) moderate, and 78(32,5%) mild. There was no statistical difference among them in MMSE or in the verbal fluency test. However, comparing to mild, patients with advanced CKD presented significantly worse cognitive performance measured by TMTA [50,8s ± 31.1s versus 66,6s ± 35,7s (p = 0.016)] and TMTB [92,7s ± 46,2s versus 162,4s ± 35,7s (p < 0.001)]. Significantly lower TMTB scores (CI95%) 33,0s (4,5-61,6s) were observed in patients with mild compared to advanced CKD in the multivariate analysis adjusting for age, education, sex, diabetes, and alcohol use. CONCLUSION: Advanced CKD is independently associated with poorer cognitive performance measured by an executive performance test compared to mild CKD.