Antibacterial Activity of Lactic Acid Bacteria to Improve Shelf Life of Raw Meat.

Lactic Acid Bacteria （LAB） are generally recognized as safe. It has been used to increase the shelf-life of fermented products, and its antimicrobial action is based on the metabolites secretions, such as lactic acid, hydrogen peroxide, reuterin, bacteriocins and the like-bacteriocins substances. It has been proven that LAB are able to inhibit deteriorating bacteria of raw meat, but improper handling of live cultures could lead to spoilage. So, the use of their bacteriocins, small antimicrobial peptides, could be an alternative. Besides reducing the number of spoilage bacteria, it seeks to inhibit pathogenic bacteria such as Salmonella, enterohemorrhagic Escherichia coli and Listeria. The food industry uses few bacteriocins and now bacterial resistance has been reported. For that reason, the search of novel bacteriocins produced by LAB is a priority. Moreover, the natural microbiota of meat could be a reservoir of LAB.

Serial monitoring of isavuconazole blood levels during prolonged antifungal therapy.

BACKGROUND: Isavuconazole is the newest triazole antifungal approved for the treatment of invasive aspergillosis (IA) and invasive mucormycosis in adult patients. OBJECTIVES: To characterize the assessment of the blood levels of isavuconazole and their association with efficacy and toxicity. METHODS: From January 2017 to May 2018, blood samples obtained from patients receiving isavuconazole were analysed for therapeutic drug monitoring. Factors influencing the blood concentrations of isavuconazole, such as weight, length of treatment, route of administration and results of selected liver function tests, were analysed in univariate and multivariate models. The receiver operating characteristic (ROC) curve was analysed to detect the best cut-off for isavuconazole toxicity. RESULTS: A total of 264 isavuconazole blood concentrations in 19 patients were analysed. The median value of isavuconazole concentration in all patients during the first 30 days of therapy was 3.69 mg/L (range 0.64-8.13 mg/L). A linear increase of 0.032 mg/L (range 0.023-0.041 mg/L) for each day of treatment (P = 0.002) was observed. In multivariate analysis the association between the length of treatment and higher levels of isavuconazole (P < 0.001) and higher serum GGT and lower isavuconazole levels (P = 0.001) was confirmed. Adverse events, mainly gastrointestinal, were reported in six patients (31.6%). Based on time-dependent and fixed-time ROC curve analysis, 4.87 mg/L and 5.13 mg/L, respectively, were the identified thresholds for toxicity. CONCLUSIONS: Isavuconazole was efficacious and well tolerated. Side effects, mainly gastrointestinal, were associated with prolonged administration and high serum levels.

Adenoid cystic carcinoma with myoepithelial predominance affecting maxilla and maxillary sinus.

Adenoid cystic carcinoma (ACC) represents less than 10% of all salivary gland tumors, rarely occurring centrally in the jaws. Herein we describe the case of a 36-year-old female patient presenting a painless swelling in the right maxilla, resulting in marked facial asymmetry. Intra-orally it was observed a swelling covered by an erythematous and irregular-surfaced mucosa, affecting the gingiva and crossing the midline of the hard palate. Imaging studies showed a mixed radiolucent-radiopaque lesion with ill-defined borders, involving the right side of the maxilla and the maxillary sinus. Incisional biopsy revealed basophilic cribriform tumoral islands and solid sheets of neoplastic cells invading bone trabeculae. Most of the tumoral cells presented myoepithelial characteristics, while few true luminal/epithelial cells were observed. To illustrate the epithelial-myoepithelial pattern, immunohistochemical reactions were performed, as well as double immunohistochemical staining. The diagnosis was intraosseous ACC, which features were discussed as well as the potential differential diagnosis.

Gamma radiation treatment applied to microbial decontamination of products derived from eggs collected from the retail market in São Paulo / Tratamento por radiação gama aplicado na descontaminação microbiana de produtos derivados de ovos coletados no mercado varejista da cidade de São Paulo

The quality of products derived from eggs depends on the quality of the raw material source and the industrial production. The contamination by fungi and bacteria can occur after exposure of the product in the retail market. The objective of this study was to evaluate the microbiological quality of 60 samples of derivated egg products and evaluate the effects of gamma radiation of cobalt-60, using the dose of 5 and 10 kGy, for decontamination of products collected in the retail market in the São Paulo city. The bacterial count was performed by the most probable number (MPN) and the fungal counts by serial dilution after plating surface. It was observed a decrease in the humidity and water activity (Aw) values of irradiated whole egg powder and white egg powder samples (p<0.05), but there was no significant change in the pH of the same samples irradiated with 5 and 10 kGy (p>0.05). In the powdered yolk the values of pH and Aw presented a significant difference (p<0.05), but no significant difference in the humidity values of 5 and 10 kGy irradiated samples (p>0.05). Effective microbial sterilization of all products occurred at the dose of 10 kGy.(AU)

A qualidade dos produtos derivados de ovos depende da qualidade da matéria-prima e da produção industrial. A contaminação por fungos e bactérias pode ocorrer após a exposição do produto no mercado varejista. O objetivo deste estudo foi avaliar a qualidade microbiológica de 60 amostras de derivados de ovos e avaliar os efeitos da radiação gama de cobalto-60, utilizando-se as doses de 5 e 10 kGy, para descontaminação dos produtos coletados no mercado varejista da cidade de São Paulo. A contagem bacteriana foi realizada pelo número mais provável (MPN), e as contagens fúngicas por diluição seriada em plaqueamento de superfície. Foi observada uma diminuição dos valores da umidade e da atividade de água (Aa) das amostras irradiadas de ovo integral em pó e clara em pó (p<0,05), mas sem alteração significativa do pH nas mesmas amostras irradiadas com 5 e 10 kGy (p>0,05). Em gema em pó, observou-se diferença significativa (p<0,05) nos valores de pH e Aa, mas não houve mudança expressiva nos valores de umidade nas amostras irradiadas com 5 e 10 kGy (p>0,05). A esterilização microbiana eficaz de todos os produtos ocorreu com a dose de 10 kGy.(AU)

Waardenburg syndrome type I: Dental phenotypes and genetic analysis of an extended family.

BACKGROUND: The aim of this study was to describe the pattern of inheritance and the clinical features in a large family with Waardenburg syndrome type I (WS1), detailing the dental abnormalities and screening for PAX3 mutations. MATERIAL AND METHODS: To characterize the pattern of inheritance and clinical features, 29 family members were evaluated by dermatologic, ophthalmologic, otorhinolaryngologic and orofacial examination. Molecular analysis of the PAX3 gene was performed. RESULTS: The pedigree of the family,including the last four generations, was constructed and revealed non-consanguineous marriages. Out of 29 descendants, 16 family members showed features of WS1, with 9 members showing two major criteria indicative of WS1. Five patients showed white forelock and iris hypopigmentation, and four showed dystopia canthorum and iris hypopigmentation. Two patients had hearing loss. Dental abnormalities were identified in three family members, including dental agenesis, conical teeth and taurodontism. Sequencing analysis failed to identify mutations in the PAX3 gene. CONCLUSIONS: These results confirm that WS1 was transmitted in this family in an autosomal dominant pattern with variable expressivity and high penetrance. The presence of dental manifestations, especially tooth agenesis and conical teeth which resulted in considerable aesthetic impact on affected individuals was a major clinical feature. CLINICAL RELEVANCE: This article reveals the presence of well-defined dental changes associated with WS1 and tries to establish a possible association between these two entities showing a new spectrum of WS1.

Evaluation of EPS extraction protocols from anaerobic sludge for gel-based proteomic studies.

Despite the importance of anaerobic sludge extracellular polymeric substances (EPSs), their characterization is limited to information regarding their chemical classes and molecular size. This work explores the possibility of using proteomic techniques to study the proteins present in this matrix. Thus, this paper compares eight EPS extraction methods regarding extraction yield, protein/carbohydrate ratio, size distribution profile and suitability to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses. Despite the differences found in quantification and size exclusion chromatography assays, the band profile found for all methods was very similar. Considering the band pattern, extraction time and background level, heating method followed by ammonium sulfate precipitation proved to be the most appropriate method for gel-based analyses of anaerobic sludge EPS proteins.

PREPARATION OF B-CYCLODEXTRIN-THIABENDAZOLE-PIPERONYL BUTOXIDE SUPRAMOLECULAR COMPLEX AND ITS ACTIVITY AGAINST BLUE AND GREEN MOULD DECAY ON INOCULATED SATSUMA FRUIT.

The supramolecular complex of ß-cyclodextrin-thiabendazole-piperony butoxide (ßCD-TBZ/PBO) was prepared and its structure was characterized by 1H NMR. Additionally, the antifungal activity of ßCD-TBZ/PBO was investigated in comparison with the commercially available thiabendazole (TBZ) fungicide by in vitro tests and on artificially inoculated 'Okitsu' satsuma fruit dipped in water at 20 degrees C or at 50 degrees C to control postharvest blue (Penicillium italicum) and green mould (P. digitatum). ß-CD-TBZ/PBO is stable for several months when stored as powder in a dark bottle. At pH 7.0 the water solubility of the ßCD-TBZ/PBO complex was consistently higher than free TBZ. Water dip at 20 degrees C did not affect decay incidence caused by blue mould but favoured the development of green mould during 4-8 days of storage at 20 degrees C with respect to untreated (control) fruit. Water at 50 degrees C effectively reduced the incidence of blue mould and totally suppressed green mould during the first 4 days but lost its efficacy afterwards. By contrast, both TBZ and ßCD-TBZ/PBO had a lasting effect and were equally effective in controlling green and blue mould decay when applied at 20 degrees C and 60 mg L(-1) active ingredient (a.i.). When applied at 50 degrees C and 20 mg L(-1) a.i. the activity of the complex against blue mould was inferior than the corresponding treatment with TBZ. In vitro assays revealed a significant effectiveness of ßCD-TBZ/PBO complex at low concentration compared to commercial formulation of TBZ.

Atividade antimicrobiana in vitro de extratos de plantas do bioma caatinga em isolados de Escherichia coli de suínos / Antimicrobial activity in vitro extracts from caatinga plants in isolated Escherichia coli from pigs

RESUMO A suinocultura é uma atividade pecuária bem consolidada no Brasil. Por outro lado a colibacilose neonatal, cujo patógeno é Escherichia coli, pode diminuir a produtividade nas granjas e causar prejuízos aos produtores. O tratamento baseia-se na utilização de drogas antimicrobianas. Todavia, o uso indiscriminado dessas substâncias tem levado a seleção de cepas resistentes. Diante disso, a busca por alternativas terapêuticas, como as plantas medicinais, tem se tornado cada vez mais comum. Dessa maneira, objetivou-se determinar a atividade antimicrobiana de cinco extratos etanólicos de plantas do bioma caatinga: Amburana cearensis (Fr. Allem) A.C. Smith, Encholirium spectabile Mart., Hymenaea courbaril L, Neoglaziovia variegata Mez e Selaginella convoluta Spring frente a 43 isolados de Eschericha coli coletados de suínos. Para o teste de sensibilidade in vitro foi realizada a técnica da Concentração Bactericida Mínima (CBM) pelo método da microdiluição em microplaca. Os extratos apresentaram atividade antimicrobiana nas seguintes médias 138,75 175,28, 128,36, 127,71 e 129,33 μg/mL, respectivamente. Essa atividade antibacteriana pode estar relacionada a ação de metabólitos secundários presentes nos extratos dessas plantas. Dessa forma, nosso estudo pode contribuir para o desenvolvimento de alternativas terapêuticas no tratamento de infecções, como a colibacilose neonatal em suíno, bem como para o conhecimento acerca das plantas medicinais da Caatinga.

ABSTRACT Swine production is a well-established livestock activity in Brazil. On the other hand, the Neonatal Colibacillosis, whose pathogen is Escherichiacoli, can decrease the productivity on farms and cause losses to producers. The treatment of the disease is based on the use of antimicrobial drugs. However, the free use of these substances has led to the selection of resistant strains. Thus, the search for alternative therapies such as medicinal plants has become becoming increasingly common. In this context, we aimed to determine the antimicrobial activity of ethanol extracts of five plants from the caatinga biome: A. cearensis (Fr. Allem) AC Smith, Encholirium spectabile Mart, Hymenaea courbaril L, Neoglaziovia variegata Mez and Selaginella convoluta Spring in face of isolates of Eschericha coli collected from pigs. For the in vitro susceptibility testing, the method of Minimum Bactericidal Concentration (MBC) was chosen The extracts showed antimicrobial activity in the following averages 138.75 175.28, 128.36, 127.71 and 129.33 mg / mL, respectively. This antibacterial activity could be related to the action of secondary metabolites in the extracts of these plants. Thus, the current study can contribute to the development of alternative therapies for the treatment of infections such as swine Colibacillosis Neonatal, as well as to the knowledge of Caatinga medicinal plants.

Microbial dynamics during azo dye degradation in a UASB reactor supplied with yeast extract

The present work aimed to investigate the microbial dynamics during the anaerobic treatment of the azo dye blue HRFL in bench scale upflow anaerobic sludge bed (UASB) reactor operated at ambient temperature. Sludge samples were collected under distinct operational phases, when the reactor were stable (low variation of color removal), to assess the effect of glucose and yeast extract as source of carbon and redox mediators, respectively. Reactors performance was evaluated based on COD (chemical oxygen demand) and color removal. The microbial dynamics were investigated by PCR-DGGE (Polimerase Chain Reaction - Denaturing Gradient of Gel Electrophoresis) technique by comparing the 16S rDNA profiles among samples. The results suggest that the composition of microorganisms changed from the beginning to the end of the reactor operation, probably in response to the presence of azo dye and/or its degradation byproducts. Despite the highest efficiency of color removal was observed in the presence of 500 mg/L of yeast extract (up to 93%), there were no differences regarding the microbial profiles that could indicate a microbial selection by the yeast extract addition. On the other hand Methosarcina barkeri was detected only in the end of operation when the best efficiencies on color removal occurred. Nevertheless the biomass selection observed in the last stages of UASB operation is probably a result of the washout of the sludge in response of accumulation of aromatic amines which led to tolerant and very active biomass that contributed to high efficiencies on color removal.

Terpinen-4-ol and alpha-terpineol (tea tree oil components) inhibit the production of IL-1ß, IL-6 and IL-10 on human macrophages.

OBJECTIVE: Tea tree oil (TTO) is an essential oil with anti-inflammatory properties, steam distilled from the plant Melaleuca alternifolia. We investigated the immunomodulatory properties of TTO and its components (terpinen-4-ol and alpha-terpineol) using lipopolysaccharide (LPS)-stimulated macrophages. METHODS: The ability of TTO, terpinen-4-ol and alpha-terpineol to modulate the macrophage response to bacterial LPS stimulation was assessed by ELISA for tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6 and IL-10 cytokine production and by western blotting for the activation of nuclear factor kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signaling, which are associated with the expression of pro-inflammatory cytokines. We used a human monocytic cell line (U937) differentiated into macrophages. RESULTS: LPS induced the production of all cytokines, and TTO and its components significantly reduced the production of IL-1ß, IL-6 and IL-10. The production of TNF-α was not affected by either TTO or its major components. The modulation of cytokine production was not mediated by changes in NF-κB or p38 MAPK activation. CONCLUSION: TTO, terpinen-4-ol and alpha-terpineol can suppress the production of inflammatory mediators in LPS-stimulated human macrophages; this inhibition was mediated by interfering with the NF-kB, p38 or ERK MAPK pathways.

Clinicopathological prognostic factors of oral tongue squamous cell carcinoma: a retrospective study of 202 cases.

Although several histopathological parameters and grading systems have been described as predictive of the treatment response and outcome of oral squamous cell carcinoma (OSCC), none is universally accepted. A new scoring system, the histological risk model, was recently described to be a powerful predictive tool for recurrence and overall survival in OSCC. The aim of this study was to verify the predictive role of the histological risk model in a cohort of 202 patients at all stages of oral/mobile tongue squamous cell carcinoma (OTSCC). Demographic and clinical data were collected from the medical records and the tumours were evaluated using the histological risk model. Statistical analyses were performed using the χ(2) test, the Kaplan-Meier method, and the Cox regression model. The histological risk model showed no statistical correlation with demographic or clinical parameters and did not Predict the outcome of the OTSCC patients. However, multivariate regression analysis revealed a significant correlation of the clinical disease stage with the disease outcome. Despite major efforts to identify new predictive parameters and histological systems, clinical features are still the most reliable prognostic factors for patients with OTSCC.

Microbial dynamics during azo dye degradation in a UASB reactor supplied with yeast extract.

The present work aimed to investigate the microbial dynamics during the anaerobic treatment of the azo dye blue HRFL in bench scale upflow anaerobic sludge bed (UASB) reactor operated at ambient temperature. Sludge samples were collected under distinct operational phases, when the reactor were stable (low variation of color removal), to assess the effect of glucose and yeast extract as source of carbon and redox mediators, respectively. Reactors performance was evaluated based on COD (chemical oxygen demand) and color removal. The microbial dynamics were investigated by PCR-DGGE (Polimerase Chain Reaction - Denaturing Gradient of Gel Electrophoresis) technique by comparing the 16S rDNA profiles among samples. The results suggest that the composition of microorganisms changed from the beginning to the end of the reactor operation, probably in response to the presence of azo dye and/or its degradation byproducts. Despite the highest efficiency of color removal was observed in the presence of 500 mg/L of yeast extract (up to 93%), there were no differences regarding the microbial profiles that could indicate a microbial selection by the yeast extract addition. On the other hand Methosarcina barkeri was detected only in the end of operation when the best efficiencies on color removal occurred. Nevertheless the biomass selection observed in the last stages of UASB operation is probably a result of the washout of the sludge in response of accumulation of aromatic amines which led to tolerant and very active biomass that contributed to high efficiencies on color removal.

First Report of Asiatic Brown Rot Caused by Monilinia polystroma on Peach in Italy.

Monilinia spp. are well-known pathogens causing brown rot of fruit trees in many fruit production areas worldwide. In Italy, three Monilinia species are particularly significant with regard to fruit trees, causing blossom and twig blight and brown rot in fruits: Monilinia laxa (Aderhold and Ruhland) Honey, M. fructicola (Winter) Honey, and M. fructigena (Aderhold and Ruhland). In 2009, a new species, M. polystroma, was distinguished from M. fructigena based on morphological and molecular characteristics in Europe (3). M. polystroma is not known to occur in Italy and to date has been reported from the Czech Republic (1), Hungary (3), Poland (4), Serbia (5), and Switzerland (2). In July 2013, during a survey for fungal postharvest pathogens, stored peaches (Prunus persica (L.) Batsch) belonging to different cultivars showing brown rot symptoms were observed in the Emilia Romagna and Sardinia regions of Italy. Typical decay spots were circular and brown, tending toward black, and 5% of peaches presented a large number of yellowish or buff-colored stromata and firm decayed tissues, the symptoms originated by M. polystroma. The pathogen was isolated on V8 agar (V8A) and culture plates were incubated at 25°C in darkness for 5 days. A conidial suspension was spread on malt extract agar (MEA) and single spores were selected. M. polystroma colonies grown on potato dexstrose agar (PDA) were yellowish in color. Irregular black stromatal crusts occurred on the edges of the colonies after 10 to 12 days of incubation and on the margin was present sporogenous tissue slightly elevated above the colony surface, color buff/pale luteous (1). The conidia were one-celled, ovoid or limoniform, smooth and hyaline, and 12 to 20 × 8 to 12 µm in distilled water when grown on V8A at 22°C. The ribosomal ITS1-5.8S-ITS2 region was PCR-amplified from genomic DNA obtained from mycelium using primers ITS1 and ITS4. A BLAST search in GenBank revealed the highest similarity (99%) to M. polystroma sequences (GenBank Accession No. GU067539). Pathogenicity was confirmed using surface-sterilized mature 'Red Heaven' peaches. The fruits were wounded (2 × 2 × 2 mm) twice with a sterile needle and inoculated with 2-mm plugs of 7-day-old mycelia from fungal colony margins. The sample unit was represented by 10 fruits. Control fruits were inoculated with PDA. After 7 days of incubation at 20°C in plastic containers with high humidity, typical symptoms of brown rot developed on both the wounds of all inoculated fruits, while control fruits remained symptomless. By the 14th day, all fruits had rotted and the yellowish exogenous stromata appeared on the surface of infected peaches. The fungus isolated from inoculated fruit exhibited the same morphological and molecular features of the original isolates; the molecular analysis performed using the primers by Petroczy (3) confirmed the result of the PCR with ITS1 and ITS4 primers. To our knowledge, this is the first report of M. polystroma on peach in Italy. This is relevant because the new pathogen could spread into other European countries that are main peach producers (such as Spain), causing economic losses. Bringing it to the attention of the scientific community allows the arrangement of research studies for assessing potential resistances with a significant impact on disease control management. Further studies are necessary to determine geographic distribution, prevalence, and economic importance of this organism in Italy. References: (1) EPPO Reporting Service. 2011/134: First reports of Monilinia polystroma in Hungary and the Czech Republic. No. 6, 2011. (2) M. Hilber-Bodmer et al. Plant Dis. 96:146, 2012. (3) M. Petroczy and L. Palkovics. Eur. J. Plant Pathol. 125:343, 2009. (4) A. Poniatowska et al. Eur. J. Plant Pathol. 135:855, 2013. (5) M. Vasic et al. Plant Dis. 97:145, 2013.

Degradation of a model azo dye in submerged anaerobic membrane bioreactor (SAMBR) operated with powdered activated carbon (PAC).

This work investigated the anaerobic degradation of the model azo dye Remazol Yellow Gold RNL in an upflow anaerobic sludge blanket reactor (UASB) and two submerged anaerobic membrane (SAMBR) bioreactors, one of which (SAMBR-1) was operated with powdered activated carbon (PAC) in its interior. The reactors were operated at 35 °C with a hydraulic retention time of 24 h in three operational phases, aimed to assess the effect of external sources of carbon (glucose) or redox mediator (yeast extract) on the removal or color and organic matter. The results showed that removal efficiencies of COD (73-94%) and color (90-94%) were higher for SAMBR-1 when compared to SAMBR-2 (operated without PAC) and UASB reactors. In addition, the presence of PAC in SAMBR-1 increased reactor stability, thereby leading to a lower accumulation of volatile fatty acids (VFA). The microfiltration membrane was responsible for an additional removal of ~50% of soluble residual COD in the form of VFA, thus improving permeate quality. On its turn, PAC exhibited the ability to adsorb byproducts (aromatic amines) of azo dye degradation as well as to act as source of immobilized redox mediator (quinone groups on its surface), thereby enhancing color removal.

Quality changes of 'Sanguinello' oranges wrapped with different plastic films under simulated marketing conditions.

Chemical and eating quality of citrus fruit changes slowly after harvest, and quality alteration is mainly due to shrinkage, loss of firmness, excessive weight loss and decay rather than a reduction of nutritional value and taste features. Film wrapping may be a suitable means to reduce transpiration and preserve market quality provided film permeability to gases does not lead to: 1) a reduction of in-package O2 partial pressure at a point that would induce anaerobic respiration; 2) an increase of CO2 concentration to toxic levels. This experiment was carried out to study quality changes of 'Sanguinello' oranges treated or not treated with 500 mg/L imazalil (IMZ) and wrapped with continuous, macro- or micro-perforated polyolefinic films. Wrapped and no-wrapped fruit were stored at 20 degrees C and 60% RH for 20 or 30 days. In-package gas composition of the macro-perforated film showed no significant difference compared to air composition, while in-package partial pressure of CO2 and O2 ranged between 4 (continuous film) and 9.8 kPa (micro-perforated films), and 14.8 (continuous film) and 5 kPa (micro-perforated films), respectively. After 30 days of storage weight loss in fruit wrapped with the macro-perforated film was (4.3%) slightly lowerthan un-packed fruit (5%), while in all other packages weight loss never exceeded 0.7%.Quality changes were quite stable over storage in all treatments, although slight but significantly lower levels of total soluble solids and ascorbic acid were detected in micro-perforated films with the lowest degree of perforation. However, the sensory analysis denoted a remarkable decrease of firmness in un-wrapped or wrapped fruit with macro-perforated film, while a moderate build-up of off-flavour, which reduced the eating quality, developed in micro-perforated films. Decay ranged between 6 and 12% in not treated fruit, with the lowest incidence detected in un-wrapped fruit, whereas differences among the different films were not significant. No decay was detected in fruit treated with IMZ. 'Sanguinello' oranges can be stored under retail conditions for a month by the only means of film wrapping without important changes in chemical, eating and marketing quality provided fruit are treated with an effective fungicide to prevent decay and that in-package gas composition is not markedly changed with respect to air.

Effect of different film packaging on microbial growth in minimally processed cactus pear (Opuntia ficus-indica).

Microorganisms are natural contaminants of fresh produce and minimally processed products, and contamination arises from a number of sources, including the environment, postharvest handling and processing. Fresh-cut products are particularly susceptible to microbial contaminations because of the changes occurring in the tissues during processing. In package gas composition of modified atmosphere packaging (MAP) in combination with low storage temperatures besides reducing physiological activity of packaged produce, can also delay pathogen growth. Present study investigated on the effect of MAPs, achieved with different plastic films, on microbial growth of minimally processed cactus pear (Opuntio ficus-indica) fruit. Five different plastic materials were used for packaging the manually peeled fruit. That is: a) polypropylene film (Termoplast MY 40 micron thickness, O2 transmission rate 300 cc/m2/24h); b) polyethylene film (Bolphane BHE, 11 micron thickness, O2 transmission rate 19000 cc/m2/24h); c) polypropylene laser-perforated films (Mach Packaging) with 8, 16 or 32 100-micron holes. Total aerobic psychrophilic, mesophilic microorganisms, Enterobacteriaceae, yeast, mould populations and in-package CO2, O2 and C2H4 were determined at each storage time. Different final gas compositions, ranging from 7.8 KPa to 17.1 KPa O2, and 12.7 KPa to 2.6 KPa CO2, were achieved with MY and micro perforated films, respectively. Differences were detected in the mesophilic, Enterobacteriaceae and yeast loads, while no difference was detected in psychrophilic microorganisms. At the end of storage, microbial load in fruits sealed with MY film was significantly lower than in those sealed with BHE and micro perforated films. Furthermore, fruits packed with micro-perforated films showed the highest microbial load. This occurrence may in part be related to in-package gas composition and in part to a continuous contamination of microorganisms through micro-holes.

Postharvest decay control of citrus fruit by preharvest pyrimethanil spray.

Preharvest infections or conidia load on fruit surface by Penicillium digitatum, P. italicum, Alternaria citri and other filamentous fungi can cause important postharvest losses of citrus fruit. Reduction in pruning frequency occurred in the last decade together with un-picked yield that eventually rots on the trees have increased the risk of postharvest decay especially when environmental conditions at picking time are favourable to pathogens' development. Sanitation procedures in the packinghouses, alternate use of postharvest fungicides with different modes of action, along with fungicide application before harvest could be an effective approach to minimize postharvest decay in citrus fruit. The present study investigated the effectiveness of a preharvest treatment with pyrimethanil (PYR), a broad spectrum fungicide, recently registered in different citrus-producing countries for postharvest treatments of citrus fruit and widely used worldwide as a preharvest treatment to control various diseases in different crops. PYR (750 mg/L) was sprayed by a hand-back sprayer at run-off on 'Fremont' mandarins. The day after the treatment, half of the trees were sprayed with a 10(4) conidial suspension of P. digitatum at run-off. Fruit were harvested following 2 or 4 weeks from treatments. Sound or either wounded 2-mm-deep and 2-mm-wide or superficial wound-scratched fruit were stored at 20 degrees C and 90% RH and inspected for decay after 1, 2 or 3 weeks of storage. In fruit harvested after 2 weeks from field treatment, PYR remarkably reduced decay development during two weeks of storage in sound fruit and in wound-scratched fruit and was fairly effective even after 4 weeks from treatment, but was ineffective in fruit wounded 2 mm deep and 2 mm wide. PYR was also effective in reducing preharvest decay incited by P. digitatum, P. italicum and Botrytis cinerea, but not by other pathogens. Results show that preharvest treatment with PYR could be a feasible approach to reduce postharvest chemical control of decay of citrus fruit.

Residue levels, persistence and effectiveness of imazalil against a resistant strain of penicillium digitatum when applied in combination with heat and sodium bicarbonate.

Green and blue molds, respectively caused by Penicillium digitatum Sacc., and P. italicum Wehmer, are the most important postharvest diseases of citrus fruit Postharvest management of these pathogens is mainly based on the application of thiabendazole (TBZ) or imazalil (IMZ) fungicides. However, their intensive and prolonged use has led to the selection of TBZ- IMZ-resistant strains of these pathogens and to a reduction of TBZ and IMZ effectiveness to control postharvest decay. However, while TBZ may become completely ineffective against TBZ-resistant strains of P. digitatum, reduction of IMZ efficacy is only partial, and an effective control of decay can still be achieved by increasing its concentration, heating the treatment-solution and/or combining IMZ with sodium bicarbonate (SBC) or other food additives or natural salts. In this study, 'Desiderio' and 'Nova' mandarins were inoculated with spores of a sensitive strain of P. digitatum to IMZ and TBZ (PDs) or with a strain of P. digitatum with double resistance to both fungicides (PDr) and immersed in IMZ or TBZ emulsions at increasing concentrations up to 1000 mg/L or in IMZ (25, 200 or 400 mg/L), SBC (0.5, 1 or 2%) or IMZ + SBC emulsions either at 20 or 40 degrees C. IMZ was superior to TBZ to control decay of 'Desiderio' mandarins incited by PDs and was also effective to control decay in fruit inoculated with PDr, while TBZ even at the highest rate was completely ineffective. In 'Desiderio' mandarins inoculated with PDs, a complete control of decay was achieved with 25 mg/L IMZ but in fruit inoculated with PDr, 25 mg/L IMZ were ineffective to control decay despite in combination with SBC at 2% a synergistic effect was detected. In contrast, a good control of decay was achieved with 400 mg/L IMZ. In 'Nova' mandarins after 1 week of incubation at 20 degrees C decay incidence in fruit dipped in 400 mg/L at 20 degrees C or 200 mg/L IMZ at 40 degrees C was almost completely inhibited, while the addition of SBC at 0.5, 1 or 2% did not improve treatments performance in fruit inoculated with PDs. However, when 'Nova' mandarins were inoculated with PDr, SBC showed a modest but significant control of decay and in combination with IMZ either at 400 mg/L and 20 degrees C or 200 mg/L and 40 degrees C, significantly improved decay control. SBC did not affect IMZ residue load in 'Valencia' oranges, whereas dipping the fruit in 400 mg/L IMZ at 20 degrees C produced similar IMZ residue load as dips at 200 mg/L IMZ at 40 degrees C. In all cases, residue levels of IMZ never exceeded 2 mg/kg, which is about 40% of the maximum residue limits (MRLs) allowed in European countries. Thus, despite the selection of IMZ-resistant strains of P. digitatum, IMZ continues to be highly effective to control green mold of citrus fruit at concentrations leaving on fruit surface residue levels below the MRLs.

Analysis of mycobiota linked to insects isolated from pet food collected in pet shops / Análise da micobiota associada à entomofauna em rações a granel para animais domésticos

Insects invade stored foods such as corn, nuts and dried fruits that are ingredients of pet food. Toxigenic fungi (Aspergillus, Penicillium and Fusarium) are also contaminants of these substrates. The insects are mechanical vectors and the infestation increases the humidity of dried food, thus promoting the fungal growth and the production of mycotoxins. The objective of this study was to analyze the mycobiota associated with insect infestation found in pet foods. Twenty samples were plated on potato agar incubated at 25º C during 7 days, for the isolation of fungi from pet food and from insects isolated from each ingredient. The insects were identified by using a dichotomous key. All samples showed fungal presence, and Sitophilus zeamais was the predominant insect with the highest frequency in the collected samples, indicating it as a vector of many fungal genera. Pet foodscomposed by a mixture of seeds and grains are subject to microbial contamination, and the insects are important mechanical vectors of spoilage and toxigenic fungi, endangering animal health.

Insetos podem infestar sementes e grãos que integram as rações destinadas à alimentação de pequenos roedores e aves domésticas como amendoim, girassol, trigo, milho, aveia e outros. Fungos toxigênicos (Aspergillus, Penicillium e Fusarium) também são contaminantes destes substratos e podem ser carreados por insetos. A deterioração da massa de sementes e grãos pela atividade dos insetos associado à presença de umidade, pode ser um facilitador para o crescimento fúngico e produção de micotoxinas. O objetivo do estudo foi o de analisar a micobiota associada às sementes, grãos e insetos em ração mista para animais domésticos. Vinte amostras foram plaqueadas em ágar-batata-dextrose incubadas a 25º C durante 7 dias para o isolamento fúngico de ração e em insetos isolados de cada componente da ração. Todas as amostras apresentaram presença de fungos e Sitophilus zeamais, inseto predominante e com maior frequencia nas amostras coletadas, demonstrando ser um vetor de vários gêneros fúngicos. Rações animais compostas de misturas de sementes e grãos estão sujeitas à contaminação microbiológica, sendo os insetos importantes vetores mecânicos de fungos deteriorantes e toxigênicos, comprometendo a saúde animal.

Occurrence of pharmaceuticals and endocrine disruptors in raw sewage and their behavior in UASB reactors operated at different hydraulic retention times.

This work investigated the occurrence of pharmaceuticals and endocrine disrupting compounds (EDCs) in raw sewage (from Belo Horizonte city, Minas Gerais state, Brazil) and assessed their behavior in demo-scale upflow anaerobic sludge blanket reactors (UASB reactors) operated at different hydraulic retention times (HRT). The dissolved concentration of the studied micropollutants in the raw and treated sewage was obtained using solid phase extraction (SPE) followed by analysis in a liquid chromatography system coupled to a hybrid high resolution mass spectrometer consisting of an ion-trap and time of flight (LC-MS-IT-TOF). The natural (estradiol) and synthetic (ethinylestradiol) estrogens were hardly detected; when present, however, their concentrations were lower than the method quantification limits. The concentrations of bisphenol A and miconazole in raw sewage were similar to that reported in the literature (around 200 ng L⁻¹ and hardly detected, respectively). The antibiotics sulfamethoxazole (median 13.0 ng L⁻¹) and trimethoprim (median 61.5 ng L⁻¹), and the other pharmaceutical compounds (diclofenac and bezafibrate, with median 99.9 and 94.4 ng L⁻¹, respectively) were found in lower concentrations when compared with reports in the literature, which might indicate a lower consumption of such drugs in Brazil. The UASB reactors were inefficient in the removal of bisphenol A, and led to an increased concentration of nonylphenol in the effluent. The anaerobic reactors were also inefficient in the removal of diclofenac, and led to a partial removal of bezafibrate; whereas, for sulfamethoxazole there seemed to be a direct relationship between the HRT and removal efficiencies. For trimethoprim the sludge retention time (SRT) seemed to play an important role, although it was only partially removed in the UASB reactors.