Traditional fermentations of the local Ecuadorian cocoa type Nacional, with its fine flavor, are carried out in boxes and on platforms for a short time. A multiphasic approach, encompassing culture-dependent and -independent microbiological analyses of fermenting cocoa pulp-bean samples, metabolite target analyses of both cocoa pulp and beans, and sensory analysis of chocolates produced from the respective fermented dry beans, was applied for the investigation of the influence of these fermentation practices on the yeast and bacterial species diversity and community dynamics during cocoa bean fermentation. A wide microbial species diversity was found during the first 3 days of all fermentations carried out. The prevailing ethanol-producing yeast species were Pichia kudriavzevii and Pichia manshurica, followed by Saccharomyces cerevisiae. Leuconostoc pseudomesenteroides (glucose and fructose fermenting), Fructobacillus tropaeoli-like (fructose fermenting), and Lactobacillus fermentum (citrate converting, mannitol producing) represented the main lactic acid bacterial species in the fermentations studied, resulting in intensive heterolactate metabolism of the pulp substrates. Tatumella saanichensis and Tatumella punctata were among the members of the family Enterobacteriaceae present during the initial phase of the cocoa bean fermentations and could be responsible for the production of gluconic acid in some cases. Also, a potential new yeast species was isolated, namely, Candida sorbosivorans-like. Acetic acid bacteria, whose main representative was Acetobacter pasteurianus, generally appeared later during fermentation and oxidized ethanol to acetic acid. However, acetic acid bacteria were not always present during the main course of the platform fermentations. All of the data taken together indicated that short box and platform fermentation methods caused incomplete fermentation, which had a serious impact on the quality of the fermented dry cocoa beans.

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Cacao (Theobroma cacao L.) breeding programmes in Ecuador have focused on obtaining high-yield clones with improved disease resistance. Cacao clones should also have photosynthetic characteristics which support increased productivity. Regarding the weather conditions at the coast of Ecuador, where most of the year there are overcasts and low air evaporative demand, there is the possibility to grow cacao without overhead shade. This study focused on the photosynthetic response at two different photosynthetic photon flux densities (PPFD) of Ecuadorian cacao clones. Seven-year old cacao clones were evaluated: eight clones of Nacional type and two commercial clones (CCN 51 and EET 103), used as controls. All clones showed an increase of 35 % on average in net photosynthetic rate (A) with increasing PPFD from the light saturation point for cacao (i.e. 400 µmol m -2 s ). Such light responsiveness in A has not been reported before. Higher A was associated with higher apparent electron transport rate, while high stomatal conductance was maintained under both PPFD conditions. Under high PPFD, low non-photochemical quenching values were found, suggesting low energy dissipation. All clones showed high maximum quantum yields of PSII (F v /F m ), suggesting the absence of damage of the photochemical system. Keywords: cacao yield, chlorophyll fluorescence, CO 2 assimilation rate, gas exchange. RESUMENLos programas de mejoramiento de cacao (Theobroma cacao L.) en Ecuador se han centrado en la obtención de clones de alto rendimiento con mayor resistencia a las enfermedades. Estos clones también deben tener características fotosintéticas que apoyen una mayor productividad. En las condiciones climáticas en la costa de Ecuador, donde la mayor parte del año hay alta densidad de nubes y baja demanda evaporativa, existe la posibilidad de cultivar cacao sin sombra. Este estudio se centró en la respuesta fotosintética de clones de cacao del Ecuador en dos diferentes densidades de flujo de fotones fotosintéticos (PPFD). Se evaluaron diez clones de cacao de siete años de edad: ocho clones de tipo Nacional recientemente desarrollados por el Instituto Nacional de investigaciones Agropecuarias, y dos clones comerciales utilizados como controles (CCN 51 y EET 103). Todos los clones de cacao mostraron un aumento del 35 % en promedio en la tasa fotosintética neta (A) con el incremento del PPFD desde el punto de saturación de luz para el cacao (400 μmol m -2 s -1 ) hasta valores altos (1000 μmol m -2 s -1 ). Dicha respuesta de A a estas condiciones de luz alta no se ha

Giardia, a flagellated protozoan that infects the upper small intestine of its vertebrate host, is the most common parasitic protist responsible for diarrhea worldwide. Molecules released by the parasite, particularly excretory and secretory antigens, seemed to be associated with pathogenesis as well as with the expression of Giardia virulence. In the present work, we examined the effect of oral administration of Giardia intestinalis excretory and secretory antigens on systemic and local antibody response as well as on mucosal injuries in BALB/c mice. Significant titers of serum-specific immunoglobulin G1 (IgG1) and specific IgG2a were observed. Systemic and mucosal specific IgA antibodies were also recorded. A transient production of serum-specific IgE antibody and high total IgE levels were also detected, suggesting the presence in excretory and secretory proteins of factors promoting a specific IgE response. The sera of excretory and secretory antigen-treated mice recognized proteins of 50 and 58 kDa as well as electrophoretic bands of 15, 63, and 72 kDa that could support a proteinase activity. The in vitro exposure of G. intestinalis trophozoites to heat-inactivated sera from mice orally inoculated with excretory and secretory antigens induced a decrease of growth, revealing a complementindependent inhibitory activity of specific serum antibodies. Furthermore, histological evaluation performed on the small and large intestines revealed moderate to acute histological changes comparable to those observed in natural or experimental Giardia infection characterized by eosinophilic infiltration, hypercellularity, and enterocytic desquamation. The present results suggested that Giardia excretory and secretory antigens stimulate a preferential Th2 response, which is probably involved in the intestinal alterations associated with giardiasis. Giardia intestinalis (synonymous with Giardia lamblia andGiardia duodenalis) is a common causative agent of diarrhea disease occurring in humans and various mammal species (1). This parasite has a worldwide distribution, and its prevalence varies from 15 to 30% in developing countries (26). The infection has been shown to be more prevalent in children less than 3 years of age as well as in undernourished and immunocompromised hosts (5). Moreover, previous studies have also showed that Giardia infection induces a mucosal immune response to other allergens (13) and may be related to an increased incidence of urticaria and food allergies (10). The pathogenesis of giardiasis is not clearly understood, but villous atrophy and reduction of the absorptive area of the small intestine have been reported, which result from a brush border enzyme deficiency responsible for malabsorption (6). Studies in vitro and in vivo have suggested that the parasite is able to produce toxin(s) (3, 8), but the factors inducing mucosal alterations in Giardia infection have not been identified yet, and at this point they are still controversial (12). It has been suggested that excretory and secretory (E/S) ...