Chill brines used in ready-to-eat meat processing may be an important source of post-processing contamination by Listeria monocytogenes. The purpose of this study was to determine the efficacy of ultraviolet light (UV) in combination with antimicrobials to reduce L. monocytogenes in fresh and used chill brines. Three different antimicrobials were used in combination with UV; citric acid (CA, 0.2 and 0.5%), dimethyl dicarbonate (DMDC, 250 and 500 ppm), and hydrogen peroxide (HP, 2000 and 4000 ppm). For fresh brine studies, brine (8.0% w/v NaCl) was prepared and inoculated with a cocktail of three L. monocytogenes strains (approximately 6 log CFU/mL). Brine was treated with UV alone, antimicrobials alone, and combination of UV and antimicrobials. Moreover, to observe the effect of treatment temperature and brine circulation through the UV system on survival of listeriae cells, inoculated brine was circulated through the system without any treatment that served as control for all the treatments. For UV treatment, inoculated brine solution was exposed to UV in an Ultraviolet Water Treatment Unit (Model: AMD 150B/1/2T D; Aquionics Inc., Peak output: 254 nm) fitted with an inline chiller to maintain brine temperature of -1°C. Samples were withdrawn at regular intervals for 120 minutes. When L. monocytogenes population was no longer detectable via direct plating on MOX, enrichment was performed and suspect colonies were confirmed using API-Listeria. For antimicrobial-only (i.e., no UV) treatments, a specific concentration of antimicrobial was added in inoculated brine and samples were taken for 120 minutes. For the brine that received combination of UV and antimicrobial treatments, UV was turned on once a specific concentration of antimicrobial was added in inoculated brine and samples were withdrawn at regular intervals for 120 minutes. When treated with UV alone, L. monocytogenes population decreased from approximately 6 log CFU/mL to below the detection limit (i.e., 1 log CFU/mL) in 15 minutes with the reduction rate of 0.87 log CFU/mL per minute. However, cells were detectable by enrichment through 120 minutes. The highest rate of decline (0.90 log CFU/mL per minute) was achieved by the combination of UV and 500 ppm DMDC (UV+500 ppm DMDC), which was not significantly different from the reduction rates of UV and UV+0.5% CA. UV+500 ppm DMDC reduced L. monocytogenes to the detection limit in 15 minutes and the organism was not detected by enrichment after 60 minutes. Though the reduction rate of UV+0.5% CA was not significantly lower than the rate of UV+500 ppm DMDC (P>0.05), the former treatment resulted in non-detectable levels more quickly (45 minutes) than the latter (60 minutes). Thus, based on enrichment studies UV+0.5% CA was the most effective treatment in reducing the population of L. monocytogenes in fresh brine. Moreover, when brine was treated with 0.5% CA alone the population decreased to below detection limit in 15 minutes with the rate significantly lower than UV+500 ppm DMDC and UV+0.5% CA (P<0.05). However, L. monocytogenes was not detectable by enrichment from 60 minutes. To summarize, through enrichment studies we observed that UV+0.5% CA, UV+500 DMDC, and 0.5% CA Control were more effective than other treatments in reducing the listeriae population to a non-detectable level. Spent brine is recycled brine that was obtained from a frankfurter processor after its maximum usage. Results of spent brine studies showed that when brine was treated with UV+4000 ppm HP and UV+2000 ppm HP, L. monocytogenes population decreased to the detection limit in 45 minutes and was not detected by enrichment from 120 minutes. These treatments were observed to be the most effective treatments with a reduction rate of 0.12 log CFU/mL per minute. The reduction rate of some other treatments such as, UV+250 and 500 ppm DMDC, UV+0.2% and 0.5% CA, and UV alone was not significantly different from UV+4000 and 2000 ppm HP. However, the population was detected through enrichment up to 120 minutes in all other treatments. The results of these studies indicate that combinations of UV and antimicrobial may be more effective than either treatment alone (except 0.5% CA treatment) to process fresh chill brines. However, the antimicrobials and UV were less effective for controlling L. monocytgoenes in spent brine; presumably due to the presence of organic matter. / Ph. D.

The antioxidant properties of various blends of rosemary extract and tocopherols, either alone or with citric and ascorbic acid, were compared in filleted, headed and gutted, and minced rainbow trout (Oncorhynchus mykiss). The filleted and headed and gutted products were stored at -29 degrees C for twelve months, while the minced was stored at the same temperature for 24 weeks. Oxidation was measured by following changes in thiobarbituric reactive substances, conjugated diene hydroperoxides, texture, drip loss, pH, sensory evaluation, and gas chromatographic detection of aldehydes. Natural antioxidants, in particular those that contained citric and ascorbic acids, were effective at retarding the development of conjugated diene hydroperoxides and malonaldehyde (p<0.05). Furthermore, sensory evaluation indicated that treated samples were less oxidized. In subsequent studies, however, it was determined that the herbal flavor notes associated with natural antioxidants complicated the ability of the experienced panel to judge extent of oxidation. Also, using filleted samples, further consumer sensory panels indicated that after 12 months frozen storage, the treated and control samples were equally acceptable. For both the filleted and headed and gutted samples, no texture differences were noted over storage time or between control or variable treatments. When using natural antioxidant products, drip loss and pH were found unreliable predicators of oxidation or muscle degradation. / Master of Science

Chapter 2: Germination and Growth of Radish (Raphanus sativus) and Lettuce (Lactuca sativus) Exposed to Arsenite and Arsenate in Hydroponic Growth Solution Little information is available on the survival, uptake, and dry mass production of vegetable seedlings and maturing plants in arsenic enriched environments. Such information is however very important to many vegetable growers in areas of subsistent agricultural like Bangladesh or home-gardeners in closer proximity of As sources such as metal smelters. Accordingly we conducted research investigating (i) the germination and radical formation of radish and lettuce seeds at varying As (V) and As (III) concentrations and (ii) radish and lettuce plants in solution culture. Seed germination studies demonstrated that 0.1mM and 0.025mM are toxic threshold levels of As (III and V) for radishes and lettuce, respectively, while As (V) is more toxic to radish seeds than As (III). Arsenic (III and V) impacted both germination and radical development in radish seeds. For lettuce we observed that As had no impact on germination but reduced radical length significantly (p < 0.01). At most equimolar concentrations, As (III) was more toxic than As (V) in lettuce seeds (0.025 - 0.10mM As), a result contrary to those obtained in radish seeds (0.05 - 0.5mM As). The hydroponic growth studies showed that losses and increases in dry weight are a function of absorbed As and are dependent on the source of As: As (V) or As (III). Moreover, the effect of absorbed As (V) or As (III) on dry weight reductions and increases differed between root and shoot portions of the plants and are crop dependent. Tissue-As (originally solution As (V)) was more toxic at the radish root level and tissue-As (originally solution As (III)) was more toxic at the radish shoot level. Conversely for lettuce, As (III) caused reductions in dry weight, while As (V) had a stimulating effect on biomass production. Lower As (V) concentrations in plant tissue throughout the lettuce study and at low As (V) concentrations (0.02mM) in the radish study may be explained by the molar ratio of P:As of approximately 5. From a food nutrition safety standpoint, studies need to concentrate on sub-lethal levels in order to ensure the proper formation of the harvestable portion of the plant. Chapter 3: Adsorption of Arsenate (V) and Arsenite (III) on Goethite in the Presence and Absence of Soluble Organic Carbon The environmental fate of arsenic is of utmost importance as the U.S. EPA has recently proposed to tighten the arsenic drinking water standard from 50 ppb to 5 ppb. In natural systems the presence of dissolved organic carbon (DOC) may compete with As for adsorption to mineral surfaces, hence increasing its potential bioavailability. Accordingly, the adsorption of arsenate As (V) and As (III) on goethite (α-FeOOH) was investigated in the presence of either a peat humic acid (Hap), a Suwannee River Fulvic Acid (FA) (IHSS) or citric acid (CA). Adsorption edges and kinetic experiments were used to examine the effects of equimolar concentrations of organic adsorbates on arsenic adsorption. Adsorption envelopes were conducted over a pH range of 11 to 3, while the kinetic studies were conducted at pH 6.5 for As (V) and pH 5.0 for As (III). Arsenate adsorption was inhibited in the order of Hap > FA > CA while arsenite adsorption was inhibited in the order of CA > FA > Hap. Humic acid reduces As V adsorption starting at pH 9, with a maximum reduction at pH 6.5. Fulvic acid slightly inhibited As (V) adsorption starting at pH 5, and this inhibition increased with a decrease in pH. No effect was observed in the presence of CA. Arsenite adsorption is inhibited by HA starting a pH 7 and increases with a decrease in pH, while FA and CA reduce As (III) adsorption beginning at pH 8, with a continuous reduction as the pH decreases. The differential extent of As V adsorption in the presence of the organic acids suggests that the distribution and the respective densities of the abundant functional groups (phenol/ catechol OH or COO⁻) are significant in the adsorption of As (V). Furthermore, larger organic acids may hydrophobically partition to surfaces via a more favorable entropy driven reaction mechanism which may influence As (V) diffusion and its subsequent adsorption to surfaces. The decrease in As (III) adsorption is caused by its reduced affinity for the surface at pH values lower than 9, and the simultaneous increase in surface activity by the organic substances' via their COO⁻ functional groups. The results of these experiments suggests that dissolved organic carbon substances are capable of increasing the bioavailability of As in soil and water systems in which the dominant solid phase is a crystalline iron oxide. Chapter 4: Adsorption of Arsenate and Arsenite on Ferrihydrite in the Presence and Absence of Dissolved Organic Carbon (DOC) The adsorption of As (V) and As (III) on synthetic 2-line ferrihydrite in the presence and absence of a peat humic acid (Hap), Suwannee River Fulvic Acid (FA) or citric acid (CA) was investigated. Previous work with goethite has demonstrated the ability of DOC materials to reduce As (V) and As (III) adsorption. In this study, a batch technique was used to examine the adsorption of arsenic (III and V) and DOCs on ferrihydrite in the pH range from 3 to 11. The results obtained demonstrated that As (V) adsorption on ferrihydrite was reduced only in the presence of CA. Arsenate reduced the adsorption of all organic acids except Hap. Both FA and CA reduced As (III) adsorption on ferrihydrite, while Hap had no effect. Fulvic and citric acid adsorption on ferrihydrite was reduced in the presence of As (III), however, adsorption increases of FA and CA were observed at lower pH, which is consistent with a decrease in As(III) adsorption. The peat humic acid had no effect on As (III) adsorption, and we believe that the adsorption process of Hap and As (III and V) on ferrihydrite are independent of each other. The observed differences between this study and the study on goethite are believed to be an intricate function of ferrihydrite's surface characteristics, which affects the mechanisms of surface adsorption and hence the affinity of organic acids such as Hap, FA, and CA for the ferrihydrite surface. As such, the adsorption of DOCs to ferrihydrite are assumed to be energetically less favorable and to occur with a fewer number of ligands, resulting in lower surface coverage of weaker bond strength. Additional factors for the observed differences are discussed. This work demonstrates the importance of the solid phase in adsorption processes and functional group composition, as noticeable differences are observed in comparison to a crystalline Fe-oxide solid phase. / Master of Science

A hibridação somática é uma nova opção em programas de melhoramento de espécies cítricas, obtendo-se híbridos somáticos que podem manter integralmente a combinação genética de ambos os progenitores envolvidos na hibridação. Assim, o objetivo desse trabalho foi estudar a propagação e crescimento de combinações parentais de híbridos somáticos com potencial para serem utilizados como porta-enxertos e verificar possíveis resistências/tolerâncias à infecção de tronco e raízes por Phytophthora nicotianae e ao vírus da tristeza dos citros (CTV). Os híbridos somáticos utilizados nos trabalhos foram: limão 'Cravo' + laranja azeda, laranja 'Caipira' + limão 'Cravo', laranja 'Caipira' + tangerina 'Cleópatra', laranja 'Caipira' + limão 'Volkameriano', laranja 'Caipira' + limão 'Rugoso', tangerina 'Cleópatra' + limão 'Volkameriano', tangerina 'Cleópatra' + laranja azeda, limão 'Cravo' + tangerina 'Sunki', laranja 'Ruby Blood' + limão 'Volkameriano', laranja 'Rohde Red' + limão 'Volkameriano' e laranja 'Valência' + Fortunella obovata. Na propagação dos híbridos, estacas de 15 cm de comprimento foram retiradas de plantas matrizes dos respectivos híbridos somáticos e submetidas ao enraizamento em câmara de nebulização intermitente por 100 dias. Posteriormente, foram analisadas o sistema radicular das estacas e parte aérea. Em seguida, foram transplantadas para sacos plásticos, conduzidas em haste única e mantidas em casa-de-vegetação por 210 dias, realizando-se avaliações mensais em relação à parte aérea e ao final avaliações do sistema radicular. Para os ensaios de infecção por Phytophthora nicotianae e CTV, foram utilizadas mudas dos respectivos híbridos somáticos oriundos de estacas e ainda a inclusão de plantas testemunhas. Para os testes de resistência/tolerância a P. nicotianae, utilizou-se o método da agulha para o teste de infecção de tronco, sendo as lesões quantificadas após 25 dias da inoculação. Para o teste de podridão das raízes e radicelas, o substrato foi infectado com estruturas do patógeno, sendo analisados a parte aérea das plantas quinzenalmente e o sistema radicular após 60 dias da instalação do ensaio. Para o teste de tolerância dos híbridos somáticos ao CTV, adotou-se o método de união dos tecidos (enxertia), analisando-se a parte aérea das plantas mensalmente em três avaliações. No enraizamento das estacas, os híbridos laranja 'Caipira' + tangerina 'Cleópatra', laranja 'Caipira' + limão 'Volkameriano', limão 'Cravo' + tangerina 'Sunki' e laranja 'Rohde Red' + limão 'Volkameriano' apresentaram melhores resultados, sendo que os híbridos laranja 'Caipira' + limão 'Volkameriano' e laranja 'Rohde Red' + limão 'Volkameriano' se destacaram em relação ao desenvolvimento das estacas após o tranplantio. Os híbridos somáticos tangerina 'Cleópatra' + laranja azeda, limão 'Cravo' + tangerina 'Sunki', tangerina 'Cleópatra' + limão 'Volkameriano', laranja 'Ruby Blood' + limão 'Volkameriano', laranja 'Rohde Red' + limão 'Volkameriano' e laranja 'Caipira' + limão 'Volkameriano' apresentaram bons resultados em relação à redução da podridão do tronco e os híbridos somáticos tangerina 'Cleópatra' + limão 'Volkameriano', tangerina 'Cleópatra' + laranja azeda, laranja 'Caipira' + limão 'Volkameriano' e laranja 'Caipira' + limão 'Cravo' apresentaram tolerância a podridão das raízes e radicelas ocasionado por P. nicotianae. Os híbridos tangerina 'Cleópatra' + laranja azeda e laranja 'Valência' + Fortunella obovata apresentaram intolerância ao vírus da tristeza dos citros. / Somatic hybridization is a new alternative in citric species breeding, yielding somatic hybrids which may integrally keep the genetic combination of both progenitors involved in the hybridization. Thus, the objective of this work was to study the propagation and growth of somatic hybrids parental combinations with potential to be used as rootstock and to verify possible resistance/tolerance to trunk and roots infection by Phytophthora nicotianae and citrus tristeza virus (CTV). This work applied the following somatic hybrids: 'Cravo' lemon + sour orange, 'Caipira' orange + 'Cravo' lemon, 'Caipira' orange + 'Cleopatra' tangerine, 'Caipira' orange + 'Volkamerian' lemon, 'Caipira' orange + 'Rough' lemon, 'Cleopatra' tangerine + 'Volkamerian' lemon, 'Cleopatra' tangerine + sour orange, 'Cravo' lemon + 'Sunki' tangerine, 'Ruby Blood' orange + 'Volkamerian' lemon, 'Rohde Red' orange + 'Volkamerian' lemon and 'Valência' orange + Fortunella obovata. In hybrids propagation, stem cuttings of approximately 15 cm length were excised from matrix plants of respective somatic hybrids and submmited to rooting in intermitent nebulization chamber for 100 days. Later, the stem cuttings root system and air part were analyzed. In sequence, they were transplanted to plastic bags, conduced in only one hast and kept in greenhouse during 210 days, when monthly evaluations were performed concerning the air part and, in the end, root system evaluations. For the Phytophthora nicotianae and CTV infection essays, plants derived from the respective somatic hybrids originated from stem cuttings were used, as well as control plants. For the resistance/tolerance to P. nicotianae analysis, the needle method for the trunk infection test was applied, being the lesions quantified after 25 days post-inoculation. For the roots and radicels flashening test, substrate was infected with pathogen structures and the air part of the plants was analyzed at every 15 days; roots were analyzed 60 days after the essay implementation. To evaluate the somatic hybrids tolerance to CTV, it was adopted the method of tissues union (grafting), where the air part of the plants was analyzed once a month in three evaluations. On the stem cuttings rooting, the hybrids of 'Caipira' orange + 'Cleopatra' tangerine, 'Caipira' orange + 'Volkamerian' lemon, 'Cravo' lemon + 'Sunki' tangerine and 'Rohde Red' orange + 'Volkamerian' lemon presented the best results, with the hybrids of 'Caipira' orange + 'Volkamerian' lemon and 'Rohde Red' orange + 'Volkamerian' lemon being the top concerning the stem cuttings development after transplanting. The somatic hybrids of 'Cleopatra' tangerine + sour orange, 'Cravo' lemon + 'Sunki' tangerine, 'Cleopatra' tangerine + 'Volkamerian' lemon, 'Ruby Blood' orange + 'Volkamerian' lemon, 'Rohde Red' orange + 'Volkamerian' lemon and 'Caipira' orange + 'Volkamerian' lemon showed good results related to reducing the trunk flashening, and the somatic hybrids of 'Cleopatra' tangerine + 'Volkamerian' lemon, 'Cleopatra' tangerine + sour orange, 'Caipira' orange + 'Volkamerian' lemon and 'Caipira' orange + 'Cravo' lemon presented tolerance to roots and radicels flashening caused by P. nicotianae. The hybrids of 'Cleopatra' tangerine + sour orange and 'Valência' orange + Fortunella obovata showed to be intolerant to citrus tristeza virus.

A hibridação somática é uma nova opção em programas de melhoramento de espécies cítricas, obtendo-se híbridos somáticos que podem manter integralmente a combinação genética de ambos os progenitores envolvidos na hibridação. Assim, o objetivo desse trabalho foi estudar a propagação e crescimento de combinações parentais de híbridos somáticos com potencial para serem utilizados como porta-enxertos e verificar possíveis resistências/tolerâncias à infecção de tronco e raízes por Phytophthora nicotianae e ao vírus da tristeza dos citros (CTV). Os híbridos somáticos utilizados nos trabalhos foram: limão 'Cravo' + laranja azeda, laranja 'Caipira' + limão 'Cravo', laranja 'Caipira' + tangerina 'Cleópatra', laranja 'Caipira' + limão 'Volkameriano', laranja 'Caipira' + limão 'Rugoso', tangerina 'Cleópatra' + limão 'Volkameriano', tangerina 'Cleópatra' + laranja azeda, limão 'Cravo' + tangerina 'Sunki', laranja 'Ruby Blood' + limão 'Volkameriano', laranja 'Rohde Red' + limão 'Volkameriano' e laranja 'Valência' + Fortunella obovata. Na propagação dos híbridos, estacas de 15 cm de comprimento foram retiradas de plantas matrizes dos respectivos híbridos somáticos e submetidas ao enraizamento em câmara de nebulização intermitente por 100 dias. Posteriormente, foram analisadas o sistema radicular das estacas e parte aérea. Em seguida, foram transplantadas para sacos plásticos, conduzidas em haste única e mantidas em casa-de-vegetação por 210 dias, realizando-se avaliações mensais em relação à parte aérea e ao final avaliações do sistema radicular. Para os ensaios de infecção por Phytophthora nicotianae e CTV, foram utilizadas mudas dos respectivos híbridos somáticos oriundos de estacas e ainda a inclusão de plantas testemunhas. Para os testes de resistência/tolerância a P. nicotianae, utilizou-se o método da agulha para o teste de infecção de tronco, sendo as lesões quantificadas após 25 dias da inoculação. Para o teste de podridão das raízes e radicelas, o substrato foi infectado com estruturas do patógeno, sendo analisados a parte aérea das plantas quinzenalmente e o sistema radicular após 60 dias da instalação do ensaio. Para o teste de tolerância dos híbridos somáticos ao CTV, adotou-se o método de união dos tecidos (enxertia), analisando-se a parte aérea das plantas mensalmente em três avaliações. No enraizamento das estacas, os híbridos laranja 'Caipira' + tangerina 'Cleópatra', laranja 'Caipira' + limão 'Volkameriano', limão 'Cravo' + tangerina 'Sunki' e laranja 'Rohde Red' + limão 'Volkameriano' apresentaram melhores resultados, sendo que os híbridos laranja 'Caipira' + limão 'Volkameriano' e laranja 'Rohde Red' + limão 'Volkameriano' se destacaram em relação ao desenvolvimento das estacas após o tranplantio. Os híbridos somáticos tangerina 'Cleópatra' + laranja azeda, limão 'Cravo' + tangerina 'Sunki', tangerina 'Cleópatra' + limão 'Volkameriano', laranja 'Ruby Blood' + limão 'Volkameriano', laranja 'Rohde Red' + limão 'Volkameriano' e laranja 'Caipira' + limão 'Volkameriano' apresentaram bons resultados em relação à redução da podridão do tronco e os híbridos somáticos tangerina 'Cleópatra' + limão 'Volkameriano', tangerina 'Cleópatra' + laranja azeda, laranja 'Caipira' + limão 'Volkameriano' e laranja 'Caipira' + limão 'Cravo' apresentaram tolerância a podridão das raízes e radicelas ocasionado por P. nicotianae. Os híbridos tangerina 'Cleópatra' + laranja azeda e laranja 'Valência' + Fortunella obovata apresentaram intolerância ao vírus da tristeza dos citros. / Somatic hybridization is a new alternative in citric species breeding, yielding somatic hybrids which may integrally keep the genetic combination of both progenitors involved in the hybridization. Thus, the objective of this work was to study the propagation and growth of somatic hybrids parental combinations with potential to be used as rootstock and to verify possible resistance/tolerance to trunk and roots infection by Phytophthora nicotianae and citrus tristeza virus (CTV). This work applied the following somatic hybrids: 'Cravo' lemon + sour orange, 'Caipira' orange + 'Cravo' lemon, 'Caipira' orange + 'Cleopatra' tangerine, 'Caipira' orange + 'Volkamerian' lemon, 'Caipira' orange + 'Rough' lemon, 'Cleopatra' tangerine + 'Volkamerian' lemon, 'Cleopatra' tangerine + sour orange, 'Cravo' lemon + 'Sunki' tangerine, 'Ruby Blood' orange + 'Volkamerian' lemon, 'Rohde Red' orange + 'Volkamerian' lemon and 'Valência' orange + Fortunella obovata. In hybrids propagation, stem cuttings of approximately 15 cm length were excised from matrix plants of respective somatic hybrids and submmited to rooting in intermitent nebulization chamber for 100 days. Later, the stem cuttings root system and air part were analyzed. In sequence, they were transplanted to plastic bags, conduced in only one hast and kept in greenhouse during 210 days, when monthly evaluations were performed concerning the air part and, in the end, root system evaluations. For the Phytophthora nicotianae and CTV infection essays, plants derived from the respective somatic hybrids originated from stem cuttings were used, as well as control plants. For the resistance/tolerance to P. nicotianae analysis, the needle method for the trunk infection test was applied, being the lesions quantified after 25 days post-inoculation. For the roots and radicels flashening test, substrate was infected with pathogen structures and the air part of the plants was analyzed at every 15 days; roots were analyzed 60 days after the essay implementation. To evaluate the somatic hybrids tolerance to CTV, it was adopted the method of tissues union (grafting), where the air part of the plants was analyzed once a month in three evaluations. On the stem cuttings rooting, the hybrids of 'Caipira' orange + 'Cleopatra' tangerine, 'Caipira' orange + 'Volkamerian' lemon, 'Cravo' lemon + 'Sunki' tangerine and 'Rohde Red' orange + 'Volkamerian' lemon presented the best results, with the hybrids of 'Caipira' orange + 'Volkamerian' lemon and 'Rohde Red' orange + 'Volkamerian' lemon being the top concerning the stem cuttings development after transplanting. The somatic hybrids of 'Cleopatra' tangerine + sour orange, 'Cravo' lemon + 'Sunki' tangerine, 'Cleopatra' tangerine + 'Volkamerian' lemon, 'Ruby Blood' orange + 'Volkamerian' lemon, 'Rohde Red' orange + 'Volkamerian' lemon and 'Caipira' orange + 'Volkamerian' lemon showed good results related to reducing the trunk flashening, and the somatic hybrids of 'Cleopatra' tangerine + 'Volkamerian' lemon, 'Cleopatra' tangerine + sour orange, 'Caipira' orange + 'Volkamerian' lemon and 'Caipira' orange + 'Cravo' lemon presented tolerance to roots and radicels flashening caused by P. nicotianae. The hybrids of 'Cleopatra' tangerine + sour orange and 'Valência' orange + Fortunella obovata showed to be intolerant to citrus tristeza virus.

Background: Trauma is a major cause of death and disability. Adverse events, such as prolonged prehospital time, hypoxia, hypotension and/or hyperventilation have been reported to correlate to poor outcome. Adequate cortisol levels are essential for survival after major trauma. In hypotensive critically ill patients, lack of sufficient amount of cortisol can be suspected, and a concept of critical illness related corticosteroid insufficiency has been proposed. Corticosteroid therapy has many adverse effects in critically ill patients and should only be given if life-saving. Correct measurement of serum cortisol levels is important but difficult in critically ill patients with capillary leakage. Estimation of the free and biologically active cortisol is preferable. In serum less than 10% of cortisol is free and biologically active and not possible to measure with routine laboratory methods. Salivary cortisol can be used as a surrogate for free cortisol, but salivary production is reduced in critically ill patients. Liver resection could reduce cortisol levels due to substrate deficiency. Aims: 1. Evaluate the occurrence of early adverse events in patients with traumatic brain injury and relate them to outcome. 2. Assess cortisol levels over time after trauma and correlate to severity of trauma, sedative/analgesic drugs and cardiovascular function. 3. Evaluate if saliva stimulation could be performed without interfering with salivary cortisol levels. 4. Assess cortisol levels over time after liver resection in comparison to other major surgery. Results: There was no significant correlation between prehospital time ³60 minutes, hypoxia (saturation &lt;95%), hypotension (systolic blood pressure &lt;90 mmHg), or hyperventilation (ETCO2 &lt;4.5 kPa) and a poor outcome (Glasgow Outcome Scale 1-3) in patients with traumatic brain injury. Cortisol levels decreased significantly over time after trauma, but there was no correlation between low (&lt;200 nmol/L) serum cortisol levels and severity of trauma. Infusion of sedative/analgesic drugs was the strongest predictor for a low (&lt;200 nmol/L) serum cortisol. The odds ratio for low serum cortisol levels (&lt;200 nmol/L) was 8.0 for patients receiving continuous infusion of sedative/analgesic drugs. There was no significant difference between unstimulated and stimulated salivary cortisol levels (p=0.06) in healthy volunteers. Liver resection was not associated with significantly lower cortisol levels compared to other major surgery. Conclusion: There was no significant correlation between early adverse events and outcome in patients with traumatic brain injury. Cortisol levels decreased significantly over time in trauma patients. Low cortisol levels (&lt;200 nmol/L) were significantly correlated to continuous infusion of sedative/analgesic drugs. Saliva stimulation could be performed without interfering with salivary cortisol levels. Liver resection was not associated with low cortisol levels compared to other major surgery.

Com o objetivo de avaliar o efeito da cobertura morta, produzida pelos adubos verdes lab-lab (Dolichos lablab L.), guandu-anão (Cajanus cajan (L.) Millsp), crotálaria (Crotalaria juncea L.), guandu (Cajanus cajan (L.) Millsp), milheto (Pennisetum glaucum (L.) Leeke), uma mistura de três coberturas (D. lablab + C. cajan + C. juncea), e a infestação natural do pomar de laranja, composta principalmente por capim-colonião (Panicum maximum Jacq.), sobre as plantas daninhas, bem como avaliar o efeito destas coberturas sobre as seguintes plantas daninhas: poaia (Richardia brasiliensis Gomes.), capim-colonião, erva-palha (Blainvillea biaristata DC.), falsa-serralha (Emilia sonchifolia (L.) DC.), capim-colchão (Digitaria horizontalis (Retz.) Koel.), corda-de-viola (Ipomea grandifolia (Dammer.) O’Donell), picão-preto (Bidens pilosa L.), guanxuma (Sida santaremnensis H. Monteiro.) e tiririca (Cyperus rotundus (L.) Pers.), em condições de pós e pré-emergência, foram instalados dois experimentos em condições de campo e quatro em condições de casa de vegetação, na Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Departamento de Produção Vegetal, Piracicaba, S.P. O delineamento experimental foi de blocos casualizados para um experimento em campo e blocos casualizados no esquema fatorial para os demais. O manejo mecânico das diferentes vegetações, foi realizado mediante a utilização de roçadeira lateral, marca KAMAQ, modelo NINJA MAC 260 que esta desenhada para projetar o material verde cortado, sobre a projeção da copa da árvore cítrica, formando dessa maneira uma camada de cobertura morta. Os parâmetros avaliados durante a condução dos experimentos, foram os seguintes: avaliações de contagem da infestação de plantas daninhas por vaso e ou por m2 (densidade), determinações de algumas propriedades químicas do solo e avaliações de produção de biomassa das plantas daninhas. Pelos dados obtidos nas condições locais dos experimentos conduzidos pode-se observar que: a utilização de coberturas mortas produzidas pelas distintas vegetações contribui, significativamente, na redução das populações das plantas daninhas, sendo os melhores resultados obtidos com a vegetação natural e a mistura dos adubos verdes; e que a adubação em área total, independentemente da cobertura vegetal utilizada apresenta uma menor densidade de plantas daninhas nas linhas das plantas cítricas. Sendo assim pode se concluir que a utilização de adubos verdes pode contribuir de maneira significativa na redução da comunidade infestante, auxiliando de forma sustentável o manejo integrado de plantas daninhas; além de contribuir com outros benefícios inerentes a utilização dessa prática. / With the objective of evaluating the effect of the mulches produced by the cover crop Dolichos lablab, Cajanus cajan, Crotalaria juncea, Pennisetum glaucum, a mixture of three cover crops (Dolichos lablab + Cajanus cajan + Crotalaria juncea), and a natural infestation in orange tree plantations composed mostly by Panicum maximum on weeds and to evaluate the effect of these cover crop on the following weeds Richardia brasiliensis, Panicum maximum, Blainvillea biaristata, Emilia sonchifolia, Digitaria horizontalis, Ipomea grandifolia, Bidens pilosa, Sida santaremnensis and Cyperus rotundus, in conditions of pos and pre-emergence, two experiments in field conditions and four in greenhouse conditions were carried out at the Superior School of Agricultura "Luiz of Queiroz", University of São Paulo, Plant Production Department, São Paulo, Piracicaba - SP - Brazil. The experimental design used was of randomized blocks for an experiment in the field and randomized blocks in the factorial design for the others. The mechanical management of the different crops was carried out utilizing a KAMAQ lateral rotary mower Ninja Mac 260, designed to throw the cut crops down forming a layer of mulch around the citric plant. The data collected during the experiments were planta/m2 and plant/pot density, determination of some chemical conditions of the soil and weed dry biomass production in g/pot. In the carried out experiments, it could be observed that the use of mulches produced by different crops contributes significantly to reduce weeds. The best results were obtained in a natural infestation and the mixture of three cover crops. The fertilization in the whole area regardless the cover crops, presents a minor density of weeds in the lines of the citric plants. Furthermore, it can be concluded that the use of cover crops contributes widely to reduce weeds, supporting integrated management of weeds and providing with other inherent benefits.

Microrganismos endofíticos são aqueles, cultiváveis ou não, que habitam o interior da planta hospedeira sem causar danos aparentes, podendo ou não apresentar estruturas externas visíveis. Esta interação endófito-planta é intrínseca a determinadas espécies de plantas e/ou bactérias. Embora bactérias do gênero Curtobacterium sejam normalmente estudadas como fitopatógenas, este grupo vem sendo isolado como endófito de diferentes espécies vegetais, tais como citros, trevo, arroz, batata, milho, olmeiro, café e álamo. Tais bactérias vem sendo estudada no controle de doenças em pepino, batata e fumo, na promoção de crescimento, interagindo com bactérias promotoras de crescimento (PGPR) e fitopatógenas. Neste contexto, o objetivo do presente estudo foi estudar a variabilidade genética e fisiológica de Curtobacterium sp. isolados endofiticamente de citros, por meio das técnicas moleculares ARDRA, RAPD, AFLP e sequenciamento do gene 16S rRNA, além de características fisiológicas (coloração da colônia, fitopatogenicidade e perfil enzimático). Além dos isolados endofíticos de citros, foram utilizados isolados endofíticos e fitopatogênicos de diversas culturas proveniente de diferentes regiões do Brasil e outros países. Embora a coloração das colônias seja uma característica altamente variável, foi observada correlação, onde os isolados endofíticos de citros apresentaram coloração variando de alaranjado a róseo, enquanto os outros isolados analisados apresentaram coloração amarelada e creme. A análise por ARDRA possibilitou a formação de 4 ribotipos (A, B, C e D), sendo que todos os fitopatógenos foram agrupados no ribotipo D. As análises de variabilidade por meio do RAPD e AFLP permitiram a separação dos isolados bacterianos em 2 grupos principais, endófitos e fitopatógenos, além de mostrar grande variabilidade dentro do grupo de isolados endofíticos de citros. De acordo com os resultados do sequenciamento do gene 16S rRNA, os isolados endofíticos de citros foram agrupados separadamente das demais espécies analisadas. Os resultados do presente estudo demonstram que os isolados endofíticos de citros apresentam genótipo divergente, sugerindo que estes isolados possam pertencer a uma nova espécie ou subespécie de Curtobacterium. / The endophytic microorganisms are those, cultivated or not, that inhabit the interior of the plant host without causing apparent damages, presenting or not visible external structures. This interaction microorganisms-plant is specific to certain species of plants and/or bacteria. Although bacteria from genus Curtobacterium genus are normally studied as phytopathogens, this group has been isolated as endophytes from several plants, such as citrus, potato, rice, maize, coffee, elm trees, red clover and poplar. Also, this bacterium has been associated to disease biocontrol in tobacco, cucumber and potato; influencing host growth or interacting with plant growth-promoting rhizobacteria and other phytopathogenic bacteria. Therefore, the aim of this study was to evaluate the genetic and physiological variability of citrus endophytic strains of Curtobacterium by physiological traits (colony color, pathogenicity and enzymatic profile), as well as molecular techniques, such as ARDRA, RAPD, AFLP and 16S rRNA gene sequencing. In the present study endophytic and phytopathogenic strains from many different plants and regions around the world were used. Although the colony color is a high variable character, correlation was observed, since phytopathogenic strains presented yellow and ivory colors, while endophytes from citrus presented orange and pink colors. Four ARDRA profile (A, B, C, D) were observed in the evaluated population, being all phytopathogenic strains include in the ribotype D. The RAPD and AFLP analysis allocated the endophytes and phytopathogens in two different major groups, besides a high variability inside citrus endophytes cluster. According to 16S rRNA gene sequence analysis, citrus endophytic were clustered in a different group of others Curtobacterium strains. The results of the present study demonstrated that the citrus endophytic strains is a divergent genotype, suggesting that citrus endophytic strains could belong to a new Curtobacterium species or subspecies.

Esse estudo tem como hipótese que a variação sazonal da fotossíntese de laranjeira ‘Valência’ está relacionada a fatores ambientais e endógenos. A menor atividade fotossintética durante os meses frios e secos está possivelmente associada à baixa temperatura e ao acúmulo de reservas, podendo ocorrer mesmo em condições de boa disponibilidade hídrica. A alta atividade fotossintética durante os meses quentes e chuvosos é possivelmente induzida pela maior disponibilidade de recursos naturais (água, energia solar e temperatura) e maior demanda por carboidratos, haja vista que nessa ocasião ocorre fluxo vegetativo intenso. O objetivo foi testar a hipótese acima, avaliando a variação anual de algumas características fisiológicas relacionadas à fotossíntese (trocas gasosas, fluorescência da clorofila, potencial de água no ramo e conteúdo foliar de carboidratos) de laranjeiras ‘Valência’ em condições de campo, em Cordeirópolis (SP). A influência da baixa disponibilidade hídrica sazonal foi verificada comparando-se plantas em condições naturais com plantas sob irrigação. A produção e algumas características biométricas foram avaliadas. A maior atividade fotossintética de laranjeiras ‘Valência’ ocorre durante a primavera, sendo induzida pelo aumento da temperatura e disponibilidade hídrica, enquanto que a menor fotossíntese ocorre durante o inverno, quando há baixa temperatura e deficiência hídrica. Durante o inverno, a menor fotossíntese não é associada ao conteúdo total de reservas foliares. Os valores máximos de assimilação de CO2 são observados na primavera, quando as condições ambientais são intermediárias entre o verão e o inverno e há fluxos vegetativo e reprodutivo nas laranjeiras. A alta demanda de carboidratos por esses drenos causa decréscimos no conteúdo de reservas foliares entre outubro e novembro, possivelmente estimulando a fotossíntese na primavera. Embora a fotossíntese das plantas no verão seja maior que no inverno, a atividade fotossintética não alcança valores próximos aos da primavera por limitações de origem bioquímica e estomática. A menor atividade bioquímica é induzida pela maior temperatura do ar em relação à primavera, em especial durante o período vespertino, enquanto que a limitação de origem estomática é causada pela alta demanda evaporativa do ar no verão. Durante o período quente e chuvoso, a atividade fotoquímica se ajusta às condições ambientais e não limita a fixação de carbono. A menor atividade fotossintética durante o inverno é ocasionada pela menor condutância estomática, devida à baixa temperatura do ar e solo. A ocorrência de deficiência hídrica no inverno causa redução do potencial de água nos ramos, diminuindo ainda mais a condutância estomática. No inverno, há limitação bioquímica da fotossíntese devida à baixa temperatura do ar, indicada pela redução da eficiência instantânea de carboxilação. Em relação ao metabolismo fotoquímico, não há indícios de limitação da fotossíntese. / This study has as hypothesis that the seasonal variation of photosynthesis in ‘Valencia’ sweet orange plant is related to environmental and endogenous factors. The lower photosynthetic activity during cold and dry months is possibly associated to low temperature and accumulation of leaf reserves, occurring even in well-watered plants.The higher photosynthetic activity during warm and rainy months is probably induced by higher availability of natural resources (water, sunlight and temperature) and by carbohydrate demand, since there is intense vegetative flush during this period. The objective was to test the above hyphotesis, evaluating the annual variation of some physiological characteristics related to photosynthesis (gas exchange, chlorophyll fluorescence, stem water potential and leaf carbohydrate content) of ‘Valencia’ sweet orange plants under field conditions, in Cordeirópolis (SP). The influence of low seasonal water availability was verified by comparing plants under natural with irrigated plants. Fruit production and some biometric characteristics were evaluated. The highest photosynthetic activity in ‘Valencia’ sweet orange plants occurred in spring, being induced by increase of temperature and water availability, while the lowest photosynthesis occurred in winter, when there is low temperature and water deficiency. During the winter, the lowest photosynthesis is not associated to the total reserve content in leaves. The maximum CO2 assimilation values are observed in spring, when environmental conditions are intermediate between summer and winter and there are vegetative and reproductive flushes in sweet orange plants. The high demand for carbohydrates in those sinks causes reduction in leaf reserve contents between october and november, probably estimulating the photosynthesis in spring. Although the photosynthesis in summer is higher than in winter, the photosynthetic activity does not reach values close to ones observed in spring due to biochemical and stomatal limitations. The low biochemical activity is induced by higher air temperature when compared to spring, mainly during afternoon, while the stomatal limitation is caused by the high air evaporative demand in summer. During the warm and wet period, the photochemical activity adjusts itself to environmental conditions and does not limit the carbon fixation. The low photosynthetic activity during winter is caused by low stomatal conductance as a consequence of low air and soil temperature. The occurrence of water deficiency in winter causes reduction of stem water potential, which is an additional factor leading to reduced stomatal conductance. In winter, there is biochemical limitation of photosynthesis due to low air temperature, as indicated by reduced instantaneous carboxylation efficiency. Regarding the photochemical metabolism, there are not signs of limitation to photosynthesis.

Para testar a hipótese de que a desmineralização in situ das superfícies de contato enxerto-leito, e a forma como o enxerto é estabilizado ao leito, podem influenciar os mecanismos envolvidos na consolidação do enxerto, fragmentos ósseos de 10 mm de diâmetro foram removidos das metáfises proximais tibiais de 36 coelhos (Oryctolagus Cuniculus) e transplantados para uma área adjacente. Na tíbia esquerda dos animais, as superfícies de contato do enxerto e do leito hospedeiro foram desmineralizadas com ácido cítrico pH 1,0 por 3 minutos antes dos enxertos serem fixados ao leito. Na tíbia direita, o transplante do bloco ósseo não foi precedido de desmineralização. Metade dos enxertos foi imobilizada sobre o leito pela superposição de uma membrana não reabsorvível de politetrafluoretileno colada com cianoacrilato ao leito à distância da interface enxerto-leito. A outra metade dos enxertos foi fixada por um parafuso de titânio no centro do enxerto. Assim, foram formados 4 grupos de estudo: membrana (M), membrana + ácido (MA), parafuso (P) e parafuso + ácido (PA). Três animais de cada grupo forneceram espécimes para análise microscópica quantitativa e qualitativa aos 15, 30 e 45 dias de pós-operatório. A análise qualitativa demonstrou que não houve formação óssea na interface em nenhum espécime aos 15 dias e que nos demais períodos, em todos os grupos, a quantidade de tecido ósseo neoformado na interface e seu estágio de maturação aumentaram com o tempo. Ambos os métodos de fixação empregados foram eficientes em manter os enxertos em posição, porém a membrana promoveu menor reabsorção da estrutura do enxerto. A análise quantitativa computadorizada revelou que, aos 30 dias, os grupos MA e PA apresentaram maior área de formação óssea na interface (71,34 ± 12,03%; 56,74 ± 2,15% respectivamente) em relação aos grupos M e P (51,75 ± 11,02%; 43,95 ± 4,05% respectivamente) e superfícies de consolidação óssea mais extensas (93,41 ± 5,95%; 93,73 ± 4,96% respectivamente) do que os grupos sem tratamento ácido (73,49 ± 7,7%; 73,77 ± 11,77% respectivamente para M e P), sendo essas diferenças estatisticamente significantes (p<0,05). Aos 45 dias de pós-operatório, os grupos MA e PA (71,18 ± 8,9%; 58,97 ± 3,97% respectivamente) apresentaram resultados superiores aos grupos M e P (59,78 ± 11,28%; 46,08 ± 3,53% respectivamente) em relação à área de neoformação óssea na interface, porém essa diferença não foi significativa. Concluiu-se que a desmineralização ácida das superfícies contactantes nos enxertos ósseos autógenos em bloco na tíbia de coelhos promoveu a osteogênese na interface enxerto-leito e acelerou a consolidação dos enxertos. Além disso, quando o tratamento ácido foi associado ao uso de membrana como método de fixação, a consolidação e grau de reabsorção óssea foram otimizados. / In order to test the hypothesis that the demineralization \"in situ\" of contacting surfaces of bone graft/bone bed and the fixation method used for graft stabilization can influence the mechanisms involved in the consolidation of the graft, bone fragments of 10 mm in diameter were removed from the proximal tibial metaphysis of thirty-six male rabbits (Oryctolagus Cuniculus) and transplanted to an adjacent area. In the left tibia of the animals, the contacting surfaces of the graft and host bed were demineralized with citric acid pH 1.0 for 3 minutes before the grafts were fixed to the receptor bed. In the right tibia, the bone block transplantation was not preceded by demineralization. Half of the grafts were immobilized on the bone bed by a nonresorbable polytetrafluoroethylene membrane glued with cyanoacrylate adhesive to the host bed distant from the bone graft-bone bed interface. The other half of the grafts were fixed by a titanium screw in the center of the graft. Thus, four groups were formed: membrane (M), membrane + acid (MA), screw (P) and screw + acid (PA). Three animals from each group provided specimens for quantitative and qualitative microscopic analysis at 15, 30 and 45 days postoperatively. Qualitative analysis showed no significant bone formation at the interface in any specimen of the groups at 15 days and on the other periods in all groups, the amount of newly formed bone at the interface as well as the stage of bone maturation increased with time. Both fixation methods were effective in maintaining the graft in position, but the membrane resulted in less resorption of the graft. Quantitative analysis, performed by means of a computer program for image analysis, showed that at day 30, groups MA and PA, showed greater area of bone formation at the interface (71.34 ± 12.03%; 56.74 ± 2 15%) than groups M and P (51.75 ± 11.02%, 43.95 ± 4.05%) and more osseointegrated bone surfaces (93.41 ± 5.95%, 93.73 ± 4.96%) than those without acid treatment (73.49 ± 7.7%, 73.77 ± 11.77%), and these differences were statistically significant. At 45 days postoperatively MA and PA groups (71.18 ± 8.9%, 58.97 ± 3.97%) showed better results than the M and P groups (59.78 ± 11.28%, 46 , 08 ± 3.53%) compared to the area of new bone formation at the interface and osseointegrated surfaces, but these differences were not significant. It was concluded that the acid demineralization of contacting surfaces in autogenous onlay bone grafts in the tibia of rabbits promotes osteogenesis in bone graft-host bed interface and accelerates the consolidation of the grafts. Furthermore, the association of this surface treatment to the use of membrane/cyanoacrylate fixation method, optimizes the results regarding consolidation and degree of bone graft resorption.