Development and validation of an improved method for the detection of Salmonella in cinnamon bark and oregano leaves using the adsorbent beta zeolite in the pre-enrichment media.

The detection of Salmonella in spices is challenging due to the presence of antibacterial components. In this study, we evaluated the use of an adsorbent beta zeolite in pre-enrichment media to improve the recovery of Salmonella from cinnamon bark and oregano leaves. Samples (25 g) were spiked with varying levels of S. Montevideo or S. Senftenberg. After 2 weeks of stabilization at RT, betazeolite was added to cinnamon and oregano samples prior to the addition of 225 mL or 475 mL of pre-enrichment media, respectively. Detection sensitivity and rate of the test method were compared to the FDA Bacteriological Analytical Manual (BAM) method which requires the use of 2.5 L pre-enrichment broth. While Salmonella could not be detected in the test method using the reduced volume of pre-enrichment media alone, the addition of beta zeolite resulted in a positivity rate of 62% and 72.6% for cinnamon bark and oregano leaves respectively (all spike levels and both serovars combined). Furthermore, while there were differences in the LOD50 compared to the BAM method, there was no significant difference in the minimum level of detection between the betazeolite and the BAM methods. Our results demonstrate that the use of betazeolite in the pre-enrichment media offers a method with reduced media volumes without compromising on the sensitivity or efficiency of Salmonella detection in cinnamon bark and oregano leaves.

Fungal pretreatment to enhance the yield of phytochemicals and evaluation of α-amylase and α-glucosidase inhibition using Cinnamomum zeylanicum (L.) quills pressurized water extracts.

Bioactive compounds entrapped in plant materials can be effectively recovered using fungal enzymes. Cinnamomum zeylanicum Sri Wijaya (SW) and Sri Gemunu (SG) accessions and commercially available C. zeylanicum (CC) were subjected to fungal pretreatment and extracted with pressured water (PWE, 0·098 MPa). Thirteen fungal species were isolated and the substrate utilization ability of the species was tested using cellulose, pectin and lignin (indirectly). Total phenolic content (TPC, Folin-Ciocalteu method), proanthocyanidin content (PC, vanillin method) and α-amylase and α-glucosidase inhibitory potential of the extracts were evaluated. The anti-diabetic drug, Acarbose was used as the positive control. Trichoderma harzianum (MH298760) showed the highest cell lysis ability and hence was used for the microbial pretreatment process. Extracts of SW treated with T. harzianum species (Pre-SW) gave the highest percentage yield (4·08% ± 0·15%), significantly potent inhibition (P < 0·05) of α-amylase and α-glucosidase activities (IC50 57 ± 8 and 36 ± 8 µg ml-1 respectively), TPC (2·24 ± 0·02 mg gallic acid equivalent g-1 ), and PC (48·2 ± 0·4 mg of catechin equivalent g-1 ) compared to Pre-SG, Pre-CC and nontreated samples. Trichoderma harzianum treatment can enhance the hypoglycaemic properties, PC and TPC of Cinnamon extracts and provide new insights into the recovery of phytochemicals.

Optimization of Salmonella detection in garlic, onion, cinnamon, red chili pepper powders and green tea.

Salmonella is the causative agent of many outbreaks related to spice consumption. However, because of the antimicrobial properties of various spices which hinders recovery and detection, Salmonella detection in spices remains a challenge. The objective of this study was to optimize an enrichment broth for Salmonella growth in different spices and tea, in order to maintain an adequate pH and decrease the antimicrobial effects of spices during Salmonella enrichment and subsequent detection. Salmonella contaminated spice and tea dried samples were prepared and the detection of Salmonella was assessed using the developed broth and automated DNA extraction and RT-PCR. Double strength Buffered Peptone Water (BPW) was used to maintain pH, and L-cysteine and DL-serine were added to the broth to reduce the effects of antimicrobial compounds in spices. The modified enrichment broth allowed the growth of Salmonella from each spice sample. Sample to broth ratios varied from 1:9 (garlic powder, chili peppers and tea), to 1:20 (cinnamon). The pH value of each enrichment varied but remained above 4.8. The addition of L-cysteine (30 mmol/L) allowed Salmonella recovery and growth in garlic and onion samples and the addition of DL-serine (11.23 mmol/L) allowed the recovery and growth in cinnamon. The results indicated that Salmonella detection was achieved in <24 h in the modified (BPW + L-cysteine and DL-serine) enrichment broth followed by detection by RT-PCR. This protocol could allow for a more rapid, robust, and sensitive enrichment method for Salmonella in spices.

Optimization and characterization of red pigment production from an endophytic fungus, Nigrospora aurantiaca CMU-ZY2045, and its potential source of natural dye for use in textile dyeing.

Some of the most important natural pigments have been produced from fungi and used for coloring in food, cosmetics, textiles, and pharmaceutical products. Forty-seven isolates of endophytic fungi were isolated from Cinnamomum zeylanicum in northern Thailand. Only one isolate, CMU-ZY2045, produced an extracellularly red pigment. This isolate was identified as Nigrospora aurantiaca based on morphological characteristics and the molecular phylogenetic analysis of a combined four loci (large subunit and internal transcribed spacer of ribosomal DNA, ß-tubulin, and translation elongation factor 1-alpha genes). The optimum conditions for red pigment production from this fungus were investigated. The results indicated that the highest red pigment yield was observed in the liquid medium containing glucose as a carbon source and yeast extract as a nitrogen source, at a pH value of 5.0 and at 27 °C with shaking for 5 days. The crude red pigment revealed the highest level of solubility in methanol. A fungal red pigment was found to have high stability at temperatures ranging from 20 to 50 °C and pH values at a range of 5.0-6.0. Based on liquid chromatography-mass spectrometry analyses, the red pigment was characterized as bostrycin. The extracted pigment was used for the textile dyeing process. Crude fungal red pigment revealed the highest staining ability in cotton fabrics and displayed excellent fastness to washing, which showing negative cytotoxicity at the concentrations used to cell culture. This is the first report on bostrycin production from N. aurantiaca.

Veiculação de bactérias de importância alimentar em especiarias comercializadas em feira livre

O objetivo desse trabalho foi analisar a qualidade microbiológica das especiariascanela, orégano e pimenta do reino comercializadas na feira livre de Cruz das Almas no Recôncavo da Bahia, por meio da quantificação de coliformes a 45°C, Staphylococcus spp., Staphylococcus coagulase positiva, Bacillus spp., Escherichia coli, Bacillus cereus e Salmonella spp. A pimenta do reino diferiu estatísticamente (p>0,05) apresentando maior contagem quando comparado com a canela e o orégano. A prevalência de S. coagulase positiva foi de 60%, 57% e 47% para pimenta do reino moída, orégano e canela, respectivamente. E. coli e Salmonella estiveram presentes na pimenta do reino em 94% e 20%, respectivamente. A contagem de coliformes a 45ºC na pimenta foi maior (3,4 log NMP. g-1) em relação a canela (0,5 log NMP.g-1) e orégano (ausência). A pimenta do reino moída apresentou alta carga microbiana com patógenos de importância alimentar como Salmonella spp., Staphylococcus coagulase positiva e B. cereus.

Essential oil of Origanum majorana L., Illicium verum Hook. f. and Cinnamomum zeylanicum Blume: chemical and antimicrobial characterization / Óleos essenciais de Origanum majorana L., Illicium verum Hook. f. e Cinnamomum zeylanicum Blume: caracterização química e antimicrobiana

Essential oils of Origanum majorana L. (marjoram), Illicium verum Hook. f. (star-anise) and Cinnamomum zeylanicum Blume (cinnamon) were obtained by steam distillation using a modified Clevenger device. The antimicrobial activity of each oil was evaluated against the bacteria Staphylococcus aureus, Escherichia coli and the fungi Aspergillus flavus and Aspergillus parasiticus by observing their growth and/or mycelial inhibition through comparison with the standard dish (without oil). The essential oils were analyzed using a gas chromatograph coupled to a mass spectrometer for identification and coupled to a flame ionization detector for quantification. The major constituents of marjoram, star-anise and cinnamon essential oils were 4-terpineol, trans-anetole and cinnamic aldehyde, respectively. In in vitro tests, essential oils of marjoram and cinnamon promoted an inhibitory effect on the bacteria S. aureus and E. coli, while the essential oil of star-anise presented activity only against E. coli. Marjoram, star-anise and cinnamon oils were effective against the studied fungi, presenting an inhibitory effect. The minimal inhibitory concentration for the mycelial growth of A. parasiticus was 1 and 0.01 µL mL-1 for star-anise and cinnamon oils, respectively. The minimal inhibitory concentration for A. parasiticus was 0.25, 2 and 2 µL mL-1 for cinnamon, star-anise and marjoram oils, respectively.

Óleos essenciais de Origanum majorana L. (manjerona), Illicium verum Hook. f. (anis estrelado) e Cinnamomum zeylanicum Blume (canela) foram obtidos pela técnica de arraste a vapor d'água com aparelho de Clevenger modificado. Foram avaliadas as atividades antimicrobianas de cada um sobre as bactérias Staphylococcus aureus, Escherichia coli e para os fungos Aspergillus flavus e Aspergillus parasiticus, observando o crescimento e/ou inibição micelial, comparando-se estes com a placa-padrão (sem óleo). Os óleos essenciais foram analisados em cromatógrafo gasoso acoplado a espectrômetro de massa para a identificação e cromatógrafo gasoso com detector de ionização de chamas para a quantificação dos compostos. Os principais constituintes dos óleos essenciais de manjerona, anis-estrelado e canela foram o 4-terpineol, trans-anetol e aldeído cinâmico, respectivamente. Nos testes in vitro, os óleos essenciais de manjerona e canela promoveram efeito inibitório sobre as bactérias S. aureus e E. coli, enquanto o óleo essencial de anis estrelado apresentou atividade apenas frente E. coli. Os óleos de manjerona, anis estrelado e canela foram efetivos sobre os fungos estudados, apresentando efeito inibitório. A concentração mínima inibitória pra o crescimento micelial de A. parasiticus foi de 1 e 0,01 µL mL-1 para os óleos de anis-estrelado e canela, respectivamente. Enquanto a concentração mínima inibitória para A. parasiticus foi de 0,25; 2 e 2 µL mL-1 para os óleos de canela, anis-estrelado e manjerona, respectivamente.

Avaliação microbiológica de condimentos artesanais e industrializados da grande Recife, PE / Microbiological evaluation condiment artisan industrialized in Recife, PE

Os condimentos são importantes do ponto de vista microbiológico, já que quando produzidos e/ou comercializados em condições higiênicas precárias, favorecem o crescimento de microrganismos. Considerando tal fato, foram realizads análises microbiológicas em canela, colorífico, cominho e pimenta do reino artesanais e obtidos industrialmente , quanto ao Número Mais Provável (NMP) de coliformes; contagem de Staphylococcus aureus, Bacillus cereus, fungos e leveduras e pesquisa de Escherichia coli e Salmonella spp. Os resultados foram analisados através de Modelos Lineares Generalizados (GLM), com processo de Stepwise, ao nível de 5,0 por cento de probabilidade. Resultados negativos foram obtidos em todas as amostras para Staphylococcus aureus; a presença de Salmonella spp só foi confirmada em duas (2,08 por cento) amostras de condimentos industrializados (ComC e PrD), estando estes condimentos em condições sanitárias insatisfatórias e impróprios para o consumo humano; todas as amostras estavam em condições sanitárias satisfatórias de acordo com a legislação vigente para coliformes fecais; a presença de Escherichia coli foi confirmada em dez (10,42 por cento) das 96 amostras analisadas; 16 (16,67 por cento) amostras apresentavam-se contaminadas por B. cereus, estando três (3,13 por cento) dentro do padrão microbiológico e 13 (13,5 por cento) fora dos padrões aceitáveis; 40 (41,67 por cento) amostras estavam dentro dos padrões microbiológicos para fungos e leveduras, enquanto que 46 (47,92 por cento) estavam acima do limite estabelecido. Os resultados demonstraram que os condimentos, quando produzidos e/ou comercializados em condições higiênico-sanitárias insatisfatórias, podem veicular microrganismos potencialmente patogênicos.

Growth and toxigenesis of Aspergillus flavus isolates on selected spices.

None of the four Aspergillus flavus isolates studied was able to grow on wet cinnamon (5 g cinnamon + 3.3 mL distilled water) at 25 degrees C over 30 days. Fungal growth was weak on the curcumin, black pepper, and white pepper; good, but lower than on rice control, on the cumin and ginger; and similar to the control on sweet and hot paprika. No aflatoxin was detected in black or white pepper after 10 days at 25 degrees C. Cumin least inhibited aflatoxin synthesis, followed by hot paprika, cumin, sweet paprika, and ginger. The amount of aflatoxin detected in the spices at the end of the incubation period never exceeded 6.25% of that found in the rice control.

[Microbiological quality of spices consumed in Cuba]. / Calidad microbiológica de especias consumidas en Cuba.

The microbiological quality of some widely consumed spices in Cuba was evaluated by means of aerobic mesophilic microorganisms count, filamentous fungi, yeasts, coliforms, thermophilic and thermoresistant microorganisms. Salmonella spp was looked for too. Black pepper and cumin resulted the most contaminated spices with values of total count and thermoresistant microorganisms at levels of 10(6) per gram, and coliform values up to 10(5) per gram. Oregano and cinnamon showed satisfactory microbiological quality; the contamination detected in these spices was lower than 10(4) per gram. Salmonella spp and yeasts were not detected.