ABSTRACT: The main method of sugarcane smut (Sporisoriumscitamineum)control is the genetic resistance. This study assessed the reaction of Brazilian sugarcane genotypes to the smut. The design used was randomized blocks, with 41 genotypes and four replications. The experimental unit consisted of five seedlings per genotype. The genotypes were inoculated by immersing the buds in a smut spores’ suspension. For each genotype, it was obtained the disease incidence in percentage of stalk with whip and with the disease incidence data over time the area under the disease progress curve (AUDPC) were calculated for each genotype. Based in the AUDPC, three genotypes were classified as highly resistant (7.3%), eighteen as resistant (43.9%), twelve as moderately resistant (29.3%), three as moderately susceptible (7.3%) and five as susceptible (12.2%). Genotypes with higher levels of resistance to smut sugarcane can be used for integrated disease management and can be used as parents in new crosses in order to obtain progenies with higher proportion of genotypes resistant to smut.

RESUMO: O principal método de controle do carvão da cana-de-açúcar (Sporisorium scitamineum)é o controle genético. O objetivo deste estudo foi avaliar a reação de genótipos brasileiros de cana-de-açúcar ao carvão. O experimento foi conduzido em blocos casualizados, com 41 genótipos e quatro repetições. A unidade experimental foi composta por cinco mudas por genótipo. Os genótipos foram inoculados através da imersão das gemas em uma suspensão de esporos do fungo. Para cada genótipo, obteve-se a incidência da doença em percentagem de colmos com chicote de carvão e com os dados de incidência da doença ao longo do tempo foi calculada a área abaixo da curva de progresso da doença (AACPD) para cada genótipo. Baseado na AACPD, três genótipos foram classificados como altamente resistentes (7,3%), dezoito como resistentes (43,9%), doze como moderadamente resistentes (29,3%), três como moderadamente suscetíveis (7,3%) e cinco como suscetíveis (12,2%). Genótipos com os maiores níveis de resistência ao carvão da cana-de-açúcar que podem ser utilizados para o manejo integrado da doença e como genitores em novos cruzamentos para obter progênies com maior proporção de genótipos resistentes ao carvão.

ABSTRACT Root-knot nematodes (Meloidogyne spp.) can cause a reduction up to 50% in the production of sugarcane (Saccharum spp.). Genetic resistance is considered an important component in the management of nematodes. Therefore, the aim of this study was to evaluate the susceptibility of 12 sugarcane genotypes when subjected to parasitism by Meloidogyne javanica and Meloidogyne incognita under controlled conditions in a plant growth chamber. Four experiments were carried out in randomized blocks with 12 treatments and four replicates for M. javanica and three replicates for M. incognita. The treatments were the sugarcane genotypes: RB966928 (susceptibility pattern), RB026842, RB036168, RB036145, RB036065, RB966229, RB036066, RB036068, RB046209, RB036163, RB036153, and RB036059. Each repetition consisted of a 2-L pot with previously autoclaved substrate (1:1 of sand and soil), and a pre-sprouted seedling was transplanted. From pure populations of M. javanica and M. incognita, approximately 2,000 eggs and eventual J2 were inoculated per pot. After 120 days, the final population of nematodes in the root and soil was counted using an optical light microscope and Peters slide, and the reproduction factor (RF) was calculated. Plants that presented RF < 1 were considered resistant, and plants that presented RF > 1 were considered susceptible. Among the 12 genotypes evaluated, 66% were susceptible to parasitism by M. javanica. The genotypes RB046209, RB036163, RB036153, and RB036059 were classified as resistant to M. javanica parasitism. For the species M. incognita, the susceptible genotypes represented 75% of the total, and the resistant genotypes were RB036163, RB036153, and RB036059.

ABSTRACT Sugarcane-associated nematodes (Saccharum spp.) can reduce productivity up to 50%. Through the survey, it was possible to identify the main nematodes that occur in a certain region as a tool for designing the best management and control strategies. The aim of this study was to characterize the population of nematodes associated with sugarcane in the North Central, North Pioneiro and Northwest mesoregion of the state of Paraná, Brazil, quantify the nematode genera associated with the crop and identify the species of Pratylenchus and Meloidogyne. A total amount of 89 soil and root composite samples were collected in nine municipalities. Nematodes were extracted and counted in a Peters counting chamber under an optical light microscope. Morphological description followed identification keys. Pratylenchus spp. were identified by morphological characteristics; Meloidogyne spp. were identified by morphological characteristics and isoenzyme electrophoresis. Twelve genera of nematodes associated with sugarcane were identified: Pratylenchus, Meloidogyne, Helicotylenchus, Xiphinema, Mesocriconema, Trichodorus, Aphelenchus, Hoplolaimus, Tylenchus, Tylenchorhynchus, Ditylenchus, and Paratrichodorus. The genera Pratylenchus and Meloidogyne were found with the highest frequencies in the roots. Among the species of Pratylenchus, P. zeae and P. brachyurus were found, with P. zeae being the most frequent. Among the Meloidogyne species, only M. javanica was found. These results are essential to aid decision making in the management of phytonematodes, mainly in the development of new control strategies and in directing genetic breeding programs for development of sugarcane cultivars for the Paraná state.

ABSTRACT This study's objectives were to compare the occurrence of brown rust and smut in conventional and energy cane families and clones in field conditions and to understand how the new hybridizations could contribute to increasing the allele frequency of resistance against the main crop diseases in breeding populations. Disease occurrence was evaluated in two selection stages. Of the 2.400 clones constituting the material for selection in the first stage, 88 clones (« 4% of the total) were selected for the second stage. Considering the overall performance of the clones without symptoms of either disease in the evaluated families that would be possibly selected in the second stage, the percentage was at least 28% lower in the families selected for high biomass (37.33%) than of the selected conventional (57.14%) and intermediate biomass (66.67%) families. In the intermediate biomass and high biomass sugarcane families, brown rust and smut rates were higher in the first stage. In the second stage, brown rust incidence was higher in high biomass families. However, at the end of both selection phases, intermediate biomass and high biomass sugarcane clones without simultaneous brown rust and smut symptoms could be selected.

RESUMEN El objetivo de este estudio fue comparar la incidencia de roya marrón y carbón en familias y clones de caña de azúcar convencional y caña de azúcar para energía, a fin de evaluar el efecto de las hibridaciones en la frecuencia alélica de resistencia a estas enfer medades en el mejoramiento de los cultivos. La ocurrencia de las enfermedades fue observada en dos etapas de selección. En la primera etapa se evaluaron 30 familias, clasificadas según tipo de familia: cinco familias de caña de azúcar convencional (CO); trece familias de caña de azúcar para producción de energía con alta biomasa (HB) y, doce familias de caña de azúcar para producción de energía con biomasa intermedia (IB). Durante la segunda etapa, se evaluaron 88 genotipos libres de enfermedad: siete genotipos CO; 60 genotipos HB y 21 genotipos IB. En la primera fase de selección se observaron tasas más altas de carbón y roya en familias de caña de azúcar de biomasa intermedia (IB) y alta (HB). En la segunda fase de selección, la incidencia de la roya de las familias de alta biomasa (HB) fue mayor. En las familias con biomasa media y alta las tasas de roya marrón y carbón resultaron mayores en el primer estadio. En el segundo estadio, la incidencia de roya marrón fue mayor en las familias con alta biomasa. Al final de dos fases, se seleccionaron clones de caña de azúcar de biomasa intermedia y alta, sin síntomas de ninguna de las enfermedades evaluadas.

Abstract Cultivar RB036091 is an early-maturing sugarcane with a long period of industrial suitability; in the Central South region of Brazil, harvest is recommended between April and August, and the cultivar is indicated for planting on moderate to highly fertile soils. It is widely adaptable and has a high sugar yield and stability of agricultural yield.

RESUMO: Neste trabalho onze genótipos de cana-de-açúcar foram classificados em relação à sua resistência à ferrugem marrom, e onze quanto à resistência à ferrugem alaranjada. Foi realizada a inoculação no “cartucho foliar” de colmos com 165 dias, no município de Paranavaí, PR, em 2017. A avaliação foi feita aos 30 dias após a inoculação, utilizando uma escala de notas. Dos genótipos avaliados para ferrugem marrom, quatro foram classificados como suscetíveis, seis como moderadamente suscetíveis e um apresentou resistência moderada. Para a ferrugem alaranjada, três genótipos foram classificados como suscetíveis, sete como moderadamente suscetíveis e um como moderadamente resistente. A avaliação e classificação da reação de genótipos de cana-de-açúcar às ferrugens é uma ferramenta importante que auxilia em ensaios preliminares, na escolha de genótipos promissores para fases mais avançadas dos programas de melhoramento além de fornecer informações aos produtores na seleção das cultivares que serão plantadas.

ABSTRACT: In this research eleven sugarcane genotypes were classified in relation to their resistance to brown rust, and eleven to their resistance to orange rust. Artificial inoculation was carried out in the leaf whorl of 165-day-old plants in the city of Paranavaí, Paraná State, Brazil, in 2017. The evaluation was performed 30 days after inoculation, using a rating scale. Among the genotypes tested for brown rust, four were classified as susceptible, six as moderately susceptible and one presented moderate resistance. For orange rust, three genotypes were classified as susceptible, seven as moderately susceptible and one as moderately resistant. The evaluation and classification of the reaction of sugarcane genotypes to the rusts is an important tool that assist in preliminary trials and selection of promising genotypes for more advanced stages of breeding programs and provides information to producers on the choice of cultivars to be planted.

Abstract The sugarcane cultivar RB036066 is medium-maturing and has a long period of industrial suitability; in the South-Central region, harvest is recommended between June and September, and it is indicated for planting on medium to highly fertile soils. The cultivar is widely adaptable and has high sugar yield and stability of agricultural production.

Abstract The sugarcane cultivar RB036088 is late-maturing, harvested from September to November in south-central Brazil, and is recommended for soils with medium to high fertility. It stands out with continuously high sugar yield over the harvests, longevity of the ratoon plants, high tillering capacity and can be harvested mechanically.

RESUMO: O uso do solo fora da sua aptidão agrícola e a pouca cobertura vegetal na bacia hidrográfica prejudicam a qualidade da água utilizada para irrigação, podendo disseminar bactérias patogênicas do grupo coliformes. O objetivo do presente trabalho foi relacionar a qualidade da água de irrigação como uso do solo (intensidade e tipo de uso) e precipitação pluviométrica, em uma região produtora de hortaliças de São José dos Pinhais, Paraná. As coletas ocorreram mensalmente no ano de 2013, em dois reservatórios e uma nascente preservada. Após as coletas, as amostras foram refrigeradas em caixas de isopor e posteriormente procedeu-se às análises de coliformes totais e termotolerantes. Os resultados do efeito do uso do solo foram analisados em árvores de probabilidade. O maior uso do solo e a precipitação pluviométrica semanal acima da média aumentam a probabilidade dos coliformes termotolerantes estarem acima do permitido pela legislação. Para o período analisado e considerando os coliformes termotolerantes, apenas a água de um dos reservatórios estudados está de acordo com a legislação para ser utilizada para irrigação de hortaliças que são consumidas cruas. Os resultados obtidos são um indicativo para o poder público local quanto à necessidade da preservação ambiental e da conservação da qualidade da água na região.

ABSTRACT: Land use outside its agricultural potential and low vegetation cover in the watershed impair the quality of water used for irrigation and may contribute to the spread of pathogenic coliform bacteria. The objective of this study was to relate the quality of irrigation water with the intensity and type of land use and the rainfall in a vegetable-producing region of São José dos Pinhais, Paraná. Water samples were collected monthly in 2013 from two reservoirs and one preserved source. After collection, the samples were chilled in Styrofoam boxes and transported to the laboratory for analyses of the total and thermotolerant coliforms. Effect of land use was analyzed by probability estimation trees. High land use and weekly above average rainfall increased the probability of thermo tolerant coliforms exceeding the limit allowed under legislation. In regards to thermo tolerant coliforms in the analyzed period, the water from only one reservoir was in accordance with the legislation for the quality of water to irrigate vegetables that are consumed raw. Results of this study are an alert to the local government for the necessity of environmental preservation to maintain the water quality of the county.

Este estudo verificou a atuação da bactéria Bacillus amyloliquefaciens subsp. plantarum FZB42, na produção de mudas orgânicas de duas cultivares (‘Santa Clara I-5300’ e ‘Cereja 261’) de tomateiro (Solanum lycopersicum L.) mediante a inoculação nas sementes. Também se investigaram as características bacterianas relacionadas com a promoção do crescimento vegetal. Para isso, utilizou-se uma solução de FZB42 na concentração de 1x10(11) UFC/mL, com os tratamentos correspondendo a porcentagens em volume (20% de FZB42; 80% de FZB42 e Testemunha com 100% de água destilada), na proporção de 320 uL/g de sementes. O percentual de germinação não foi influenciado pelos tratamentos e verificou-se estímulo ao crescimento da parte aérea por FZB42 a 20% nas plântulas da cv ‘Cereja’. Na produção de mudas, a bactéria FZB42 a 20% promoveu o crescimento enquanto a FZB42 a 80% o reduziu. A inoculação nas duas doses aumentou os teores clorofila a, b e totais das mudas de tomateiro. A estirpe FZB42 apresentou resultados positivos para produção de compostos indólicos e sideróforos, e na dose de 20%, mostrou-se vantajosa na produção de mudas, ao aumentar a parte aérea em 47,7% na cultivar ‘Santa Clara’, e 15,5% na cv ‘Cereja’, quando comparados à testemunha.

This study we sought to evaluate the action of the bacteria Bacillus amyloliquefaciens subsp. plantarum in the production of organic seedlings of two cultivars (‘Santa Clara I-5300’ and ‘Cereja 261’) from tomato (Solanum lycopersicum L.) through inoculated seeds. And investigate bacterial characteristics related to plant growth promotion. For this, we used FZB42 solution at a concentration of 1x10(11) CFU/mL, with treatments correspond to percentages by volume (20% FZB42; 80% of FZB42 and control 100% distilled water) at a ratio of 320 uL/g seed. The germination percentage was not influenced by the treatments, and FZB42 to 20% increased the shoot in cultivar ‘Cereja’. In seedling production, FZB42 to 20% promoted growth while FZB42 to 80% decreased it. Inoculation with the two doses tested of FZB42 enhanced the contents of chlorophyll a, b and total in the tomato seedling. The FZB42 strain showed positive results for production of indole compounds and siderophore; and in a dose of 20% was more advantageous to the tomato seedling, promoting shoot growth with added of 47.7% for ‘Santa Clara’ and 15.5% for ‘Cereja’ when compared to the control.

Objetivou-se neste trabalho elaborar e validar uma escala diagramática para a ferrugem alaranjada da cana-de-açúcar e comparar a sua eficiência em avaliar a doença com a escala da ferrugem marrom. Para elaboração da escala foram coletadas 120 folhas de uma cultivar suscetível à doença, e a maior e menor severidades encontradas representaram os extremos da escala. Os níveis intermediários foram calculados respeitando a lei de Weber-Fechner. A escala foi proposta com nove níveis de severidade: 0,06; 0,14; 0,36; 0,89; 2,17; 5,18; 11,87; 24,92 e 45,00%. Oito avaliadores realizaram a validação estimando a severidade de 120 folhas em três avaliações: sem escala, com a escala da ferrugem marrom e com a escala da ferrugem alaranjada. A escala elaborada foi mais eficiente para avaliar a doença, proporcionando um coeficiente de determinação (R²) médio de 0,91 e ausência de erros constantes e sistemáticos. Com a escala da ferrugem marrom, o R² médio foi de 0,84, e três avaliadores cometeram desvios constantes. Com a escala elaborada, aproximadamente 92% das estimativas estiveram na faixa de variação de 5% do valor real, enquanto que nas avaliações sem escala e com a escala da ferrugem marrom, aproximadamente 78% e 84% das estimativas, estiveram nessa faixa, respectivamente.

The aim of this work was to elaborate and validate a diagrammatic scale to quantify the severity of orange rust of sugarcane and to compare its efficiency in assessing the disease with the scale of brown rust. For the design of the scale, 120 sugarcane leaves were collected from a susceptible cultivar and the highest and the lowest severity founded were selected as extremes of scale. The intermediate levels were calculated according to the law of Weber-Fechner. The scale was proposed with nine levels of severity: 0,06; 0,14; 0,36; 0,89; 2,17; 5,18; 11,87; 24,92 and 45,00%. The validation was accomplished by eight appraisers who estimated the severity of 120 leaves in three assessments: without scale, with scale of brown rust and with the proposed diagrammatic scale. The scale proposed was more efficient than the scale of brown rust to assess the severity of this disease, providing an average coefficient of determination (R²) of 0,91 and absence of constant and systematic errors. With the scale of brown rust, the average R² was 0,84 and three appraisers committed constant errors. Moreover, with the scale proposed, approximately 92% of the estimates of the appraisers had been in the range of variation of 5% of the value of real severity, while in the assessments without scale and with scale of brown rust, approximately 78% and 84% of the estimates were in that range, respectively.

O objetivo deste trabalho foi avaliar o efeito dos sistemas de manejo do solo e de coberturas de inverno sobre o número de propágulos de Fusarium spp. no solo, a incidência da podridão‑vermelha‑da‑raiz (PVR) e a produtividade das cultivares de soja CD 206 e FT Fênix. Foram realizados dois experimentos nos anos agrícolas de 2006/2007 e 2007/2008. Utilizou-se o delineamento experimental de blocos ao acaso, em parcelas subsubdivididas, com três repetições. Foram avaliados dois sistemas de preparo do solo: plantio direto e revolvimento do solo na profundidade de 25 cm. As coberturas de inverno utilizadas foram: aveia‑preta, com duas densidades de plantio; aveia‑preta + ervilhaca; azevém; e pousio. A incidência da doença, na safra de 2006/2007, na cultivar FT Fênix, foi menor que na CD 206. Na safra 2007/2008, não houve diferença significativa. Houve incremento na produtividade, de 125 kg ha‑1, com o solo revolvido, em comparação ao plantio direto. A cobertura com aveia‑preta + ervilhaca apresentou maior número de propágulos de Fusarium spp. no solo, na safra de 2006/2007. No entanto, no segundo ano, essa diferença não foi observada. Os sistemas de preparo do solo e as coberturas de inverno utilizadas não influenciam a incidência da PVR em cultivares de soja ou o número de propágulos de Fusarium spp. no solo. O sistema com solo revolvido proporciona aumento de produtividade da soja, no segundo ano de manejo.

The objective of this work was to assess the effect of soil management systems and winter cover crops on the number of propagules of Fusarium spp. in soil, the incidence of sudden death syndrome (SDS), and the productivity of the soybean cultivars CD 206 and FT Fênix. Two experiments were carried out in the 2006/2007 and 2007/2008 crop years. The experimental design was a randomized complete block in a split‑split plot arrangement, with three replicates. Two soil tillage systems were evaluated: no‑tillage and plowed soil at a depth of 25 cm. The soil covers used were: black oat, with two planting densities; black oat + vetch; ryegrass; and fallow. The incidence of the disease in the 2006/2007 crop year in the cultivar FT Fênix was lower than in CD 206. In the 2007/2008 crop, there was no significant difference. There was an increase in productivity, of 125 kg ha‑1, in the plowed treatment, when compared to no‑tillage. The cover with black oat + vetch showed a higher number of propagules of Fusarium spp. in soil in the 2006/2007 crop year. However, in the second year, this difference was not observed. The soil management systems and winter cover crops used do not influence the incidence of SDS in soybean cultivars or the number of Fusarium spp. propagules in soil. The plowed system provides an increase in soybean yield in the second year of management.

A maioria dos patógenos de soja é transmitida pela semente, merecendo destaque o fungo Sclerotinia sclerotiorum, o qual tem apresentando níveis preocupantes de incidência em campo, em vários estados brasileiros. O trabalho teve por finalidade verificar a eficiência de métodos de detecção do patógeno Sclerotinia sclerotiorum em sementes de soja, infectadas artificialmente em laboratório e oriundas de áreas com o histórico da incidência da doença em campo. Para tanto, foram utilizadas amostras de sementes de sete cultivares, provenientes de áreas infestadas e uma amostra de sementes inoculada artificialmente. Foram comparados os seguintes métodos de detecção recomendados pela literatura: Papel de Filtro ("Blotter Test"), a 7 ºC, 14 ºC e 20 ºC; Rolo de Papel; e Neon-S. Pelos resultados obtidos, verificou-se que os métodos estudados não apresentaram repetibilidade, sendo pouco sensíveis para a detecção do patógeno em sementes provenientes de áreas com incidência da doença; porém, foram eficientes para a detecção em sementes inoculadas artificialmente. No método de incubação em Papel de Filtro a 7 ºC observou-se menor incidência de outros fungos considerados indesejáveis durante as análises de sementes realizadas.

Most soybean pathogens are seed transmitted, deserving emphasis the fungus Sclerotinia sclerotiorum, which has been presenting worrying levels of field incidence in some soybean cropping areas in several Brazilian states. The objective of this study was to verify the efficiency of different methods for detecting S. sclerotiorum on soybean seeds artificially infected in the laboratory and from field production areas with a historical disease incidence. Seed samples of seven different cultivars collected from naturally infested fields, and one seed sample artificially inoculated in the laboratory were used. The following detection methods recommended in the literature were compared: Blotter test at 7 ºC, 14 ºC, and 21 ºC; Rolled Paper; and Neon-S. Results demonstrated that these methods showed no repeatability and had a low sensitivity for detecting the pathogen in seeds from areas with disease incidence. They were effective, however, for its detection on artificially inoculated seeds. In the Blotter test method at 7 ºC, there was a lower incidence of other fungi considered undesirable during seed analysis.

RB966928 is an early to medium maturing sugarcane cultivar and in Central-Southern Brazil it is harvested between April and June; it is recommended for planting in environments with medium to high soil fertility. It stands out with a high ratoon yield and with excellent sprouting in both plant cane and ratoon cane. It is tolerant to the major diseases of economic importance of sugarcane.

A hérnia das crucíferas causada por Plasmodiophora brassicae é uma das mais importantes doenças no cultivo de espécies de Brassicas no Brasil. Nos Municípios da Região Metropolitana de Curitiba, PR a maioria dos solos está contaminada pelo patógeno Plasmodiophora brassicae agente causal da hérnia das crucíferas, inviabilizando o cultivo de espécies de Brassicas em diversas propriedades. A calagem utilizada para elevar o pH do solo é uma das medidas de controle mais indicadas. Diante disso, o objetivo deste trabalho foi avaliar o efeito do pH do solo em diferentes concentrações de inóculo, no controle de P. brassicae. Foram utilizados quatro níveis de pH do solo: 4,3; 5,5; 6,2 e 7,3 e três concentrações de inóculo:1.2 x 10(7) ; 2.5 x 10(7); e 5 x 10(7) esporos.mL-1 e testemunha não inoculada. A suspensão de esporos foi obtida a partir de raízes de couve-chinesa com sintomas de hérnias e adicionado no colo das plantas, por ocasião do transplante das mudas de couve-chinesa (Brassica rapa var. pekinensis), para os vasos. Aos 45 dias após a inoculação, foram realizadas as avaliações. O efeito do pH sobre a severidade da doença foi mais expressivo em concentrações médias de inóculo (1.2 x 10(7) a 2.5 x 10(7) esporos.mL-1). Em concentrações elevadas de inóculo, a severidade da doença foi reduzida em pH do solo de 6.2 e 7.3. O melhor desenvolvimento das plantas, quantificado pelo acúmulo de massa seca foliar foi obtido nos tratamentos onde o solo apresentou pH 6,2 e 7,3 e com a menor concentração de inóculo.

Clubroot of the crucifers caused by Plasmodiophora brassicae is one of the most important diseases in the cultivation of Brassica species. In the Metropolitan Region of Curitiba, PR most of the soil is contaminated by the pathogen Plasmodiophora brassicae infesting agent of the clubroot of the crucifers making the cultivation of different species not feasible in many properties. Liming to increase soil pH is one of the most recommended control measures. Whereupon the objective of this work was to evaluate the effect of soil pH in different inoculums concentrations in the control of P. brassicae. Four levels of soil pH were used: 4,3; 5,5; 6,2 and 7,3 and three inoculums concentrations: 1.2 x 10(7), 2.5 x 10(7) 5 x 10(7) spores.mL-1 and non-inoculated control. The spores suspension was obtained from Chinese cabbage roots showing galls and it was added to the plant collars when the Chinese cabbage (Brassica rapa var. pekinensis) seedlings were transplanted to pots. At 45 days after inoculation the evaluations were performed. The effect of pH on the disease severity was more expressive with the average inoculums (1.2 x 10(7) to 2.5 x 10(7) spores.mL-1) With the higher inoculums concentrations the disease severity was reduced in soil pH 6,2 and 7,3. The best development of the plants assessed by the accumulation of foliar dry phytomass was obtained where the soil presented pH between 6,2 and 7,3 and with the lowest inoculum concentration.