Antimicrobial and Cytotoxic Activity of Fruit Extract from Syzygium cumini (L.) Skeels

The aim of the present study was to evaluate the antimicrobial and cytotoxic activity of the ethanolic extract of S. cumini according to the Clinical and Laboratory Standards Institute reference method (with modifications), determining the minimal inhibitory and lethal concentration. Activity against Gram-positive (Staphylococcus aureus and S. epidermidis), Gram-negative (Pseudomonas aeruginosa) and yeast of Candida sp and Cryptococcus neoformans was evaluated. The effects of the fruit extract were examined in hamster cells ovaries in concentrations ranging from 1250.0 a 4.9 mu g/ml, measuring the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium. The extract showed both bactericidal and fungicidal activity among the various microorganisms tested and the MIC ranging from 7.8 to 250 mu g/ml. The MIC, MBC and MFC should values that were similar for all the microorganisms. Cytotoxicity index of the dried extract corresponded to the concentration of 400 mu g/ml. The extract could potentially be used in topical antimicrobial products. Thus, the activity of extract was potent to bacteria and mainly to non-albicans species and C. neoformans.

Apoptosis and necrosis of polymorphonuclear leukocytes in goat milk with high and low somatic cell counts

The purpose of the present trial was to compare the percentages of necrotic and apoptotic polymorphonuclear leukocytes (PMNL) in goat milk with low and high somatic cell count (SCC). Twenty eight milk samples were collected from 20 lactating goats, determined to be negative in bacteriological examination, and divided in three groups, according to their SCC: samples with SCC lower than 500 x 10(3) cells/mL; between 500 and 1500 x 10(3) cells/mL; and higher than 1500 x 10(3) cells/m L. SCC was performed in an automatic somatic cell counter. Apoptosis and necrosis were quantified using dual-color flow cytometry with fluorescein labeled annexin-V and propidium iodide (PI). Results of the present study showed a significant positive correlation between the percentage of the viable PMNL and milk SCC(r = 0.495, P=0.008), as well as a significant negative correlation between apoptotic PMNL and milk SCC(r = -0.486, P = 0.009). Results also pointed out lower PMNL viability rates due to higher apoptosis rates in milk samples with SCC lower than 5 x 10(5) cells/mL. (C) 2011 Elsevier B.V. All rights reserved.

Microbial and sensory changes throughout the ripening of Prato cheese made from milk with different levels of somatic cells

The objective of this research was to evaluate the effects of 2 levels of raw milk somatic cell count (SCC) on the composition of Prato cheese and on the microbiological and sensory changes of Prato cheese throughout ripening. Two groups of dairy cows were selected to obtain low-SCC (<200,000 cells/mL) and high-SCC (>700,000 cells/mL) milks, which were used to manufacture 2 vats of cheese. The pasteurized milk was evaluated according to the pH, total solids, fat, total protein, lactose, standard plate count, coliforms at 45 degrees C, and Salmonella spp. The cheese composition was evaluated 2 d after manufacture. Lactic acid bacteria, psychrotrophic bacteria, and yeast and mold counts were carried out after 3, 9, 16, 32, and 51 d of storage. Salmonella spp., Listeria monocytogenes, and coagulase-positive Staphylococcus counts were carried out after 3, 32, and 51 d of storage. A 2 x 5 factorial design with 4 replications was performed. Sensory evaluation of the cheeses from low- and high-SCC milks was carried out for overall acceptance by using a 9-point hedonic scale after 8, 22, 35, 50, and 63 d of storage. The somatic cell levels used did not affect the total protein and salt: moisture contents of the cheeses. The pH and moisture content were higher and the clotting time was longer for cheeses from high-SCC milk. Both cheeses presented the absence of Salmonella spp. and L. monocytogenes, and the coagulase-positive Staphylococcus count was below 1 x 10(2) cfu/g throughout the storage time. The lactic acid bacteria count decreased significantly during the storage time for the cheeses from both low- and high-SCC milks, but at a faster rate for the cheese from high-SCC milk. Cheeses from high-SCC milk presented lower psychrotrophic bacteria counts and higher yeast and mold counts than cheeses from low- SCC milk. Cheeses from low- SCC milk showed better overall acceptance by the consumers. The lower overall acceptance of the cheeses from high-SCC milk may be associated with texture and flavor defects, probably caused by the higher proteolysis of these cheeses.

Short communication: Identification of subclinical cow mastitis pathogens in milk by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Subclinical mastitis is a common and easily disseminated disease in dairy herds. Its routine diagnosis via bacterial culture and biochemical identification is a difficult and time-consuming process. In this work, we show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows bacterial identification with high confidence and speed (1 d for bacterial growth and analysis). With the use of MALDI-TOF MS, 33 bacterial culture isolates from milk of different dairy cows from several farms were analyzed, and the results were compared with those obtained by classical biochemical methods. This proof-of-concept case demonstrates the reliability of MALDI-TOF MS bacterial identification, and its increased selectivity as illustrated by the additional identification of coagulase-negative Staphylococcus species and mixed bacterial cultures. Matrix-assisted laser desorption-ionization mass spectrometry considerably accelerates the diagnosis of mastitis pathogens, especially in cases of subclinical mastitis. More immediate and efficient animal management strategies for mastitis and milk quality control in the dairy industry can therefore be applied.

Long-Term Maternal Separation Differentially Alters Serum Corticosterone Levels and Blood Neutrophil Activity in A/J and C57BL/6 Mouse Offspring

Objectives: In this work, we searched for maternal separation effects on serum corticosterone levels and blood neutrophil activity in adult male A/J and C57BL/6 mouse offspring. Methods: 40 male A/J mice and 40 male C57BL/6 mice were divided within each strain into two groups. Mice in the maternal separation group were separated from their mothers (1 h/day) on postnatal days 0-13. Mice in the control group were left undisturbed. On postnatal day 45, blood was drawn from all mice and used to assess neutrophil activity by flow cytometry and serum corticosterone levels by radioimmunoassay. Results: The results showed that each mouse strain responded differently to maternal separation, but in both cases, serum corticosterone levels were affected. In both strains, adult mice that experienced maternal separation showed lower serum corticosterone levels than control mice. In relation to control mice kept together with their mothers, the levels of serum corticosterone were 72.7 and 36.36% lower in A/J and C57BL/6 mice submitted to maternal separation, respectively. The current findings showed that maternal separation increased neutrophil activity in mice after reaching adulthood. The observed effects, although in the same direction, differed between A/J and C57BL/6 mice. Maternal separation increased both the percentage and intensity of phagocytosis in C57BL/6 mice, but had no effects on A/J mice. Furthermore, maternal separation increased basal and propidium iodide-labeled Staphylococcus aureus-induced oxidative burst in A/J mice but did not affect oxidative burst in C57BL/6 mice. Finally, phorbol myristate acetate-induced oxidative burst increased in both strains. Conclusion: These results indicate that early maternal separation increases innate immunity, most likely by modifying hypothalamus-pituitary-adrenal axis activity. This suggests that maternal separation is a good model for stress which produces long-term neuroimmune changes whatever the animal species and strain used. Copyright (C) 2011 S. Karger AG, Basel

Heat stress impairs performance parameters, induces intestinal injury, and decreases macrophage activity in broiler chickens

Studies on environmental consequences of stress on animal production have grown substantially in the last few years for economic and animal welfare reasons. Physiological, hormonal, and immunological deficits as well as increases in animals` susceptibility to diseases have been reported after different stressors in broiler chickens. The aim of the current experiment is to describe the effects of 2 different heat stressors (31 +/- 1 and 36 +/- 1 degrees C/10 h per d) applied to broiler chickens from d 35 to 42 of life on the corticosterone serum levels, performance parameters, intestinal histology, and peritoneal macrophage activity, correlating and discussing the obtained data under a neuroimmune perspective. In our study, we demonstrated that heat stress (31 +/- 1 and 36 +/- 1 degrees C) increased the corticosterone serum levels and decreased BW gain and food intake. Only chickens submitted to 36 +/- 1 degrees C, however, presented a decrease in feed conversion and increased mortality. We also showed a decrease of bursa of Fabricius (31 +/- 1 and 36 +/- 1 degrees C), thymus (36 +/- 1 degrees C), and spleen (36 +/- 1 degrees C) relative weights and of macrophage basal (31 +/- 1 and 36 +/- 1 degrees C) and Staphylococcus aureus-induced oxidative burst (31 +/- 1 degrees C). Finally, mild multifocal acute enteritis characterized by an increased presence of lymphocytes and plasmocytes within the jejunum`s lamina propria was also observed. The stress-induced hypothalamic-pituitary-adrenal axis activation was taken as responsible for the negative effects observed on the chickens` performance and immune function and also the changes of the intestinal mucosa. The present obtained data corroborate with others in the field of neuroimmunomodulation and open new avenues for the improvement of broiler chicken welfare and production performance.

Neuroendocrine, behavioral and macrophage activity changes induced by picrotoxin effects in mice

The relevance and property of studies related to stress effects on immune function are undisputable. All studies conducted on stress-immune relationships, however, provide from physical and/or psychological stressors. Indeed, as far as it is of our knowledge brain-innate immune responses were not analyzed after anxiogenic-like drugs use. The present experiment was then undertaken to analyze the effects of picrotoxin (0.3, 0.6 and 1.0 mg/kg doses) on behavior, macrophage activity, serum corticosterone and noradrenaline (NE) levels and turnover in the brain of adult mice. Results showed that picrotoxin treatment in mice: (1) decreased motor and rearing activities in an open-field; (2) decreased the number of entries into the plus-maze open-arms and decreased the time spent in the exploration of the plus-maze open-arms; (3) decreased both motor activity and the level of holes exploration in the hole-board; (4) increased the levels of serum corticosterone in dose-dependent way; (5) increased noradrenaline (NE) and MHPG levels and NE turnover in the hypothalamus; and (6) increased Staphylococcus aureus and PMA-induced macrophage oxidative burst. However, and contrary to that reported after physical or psychological stress, this drug induced no effects on macrophage phagocytosis and NE levels and turnover in the frontal cortex. The present results are thus showing that picrotoxin induces some but not all neuro-innate immunity changes previously reported for inescapable foot-shock and psychological stressors in mice. These facts suggest that this chemical stressor triggers CNS pathways that might be somehow different from those fired by inescapable foot-shock and psychological stressors, leading to different neuro-innate immune responses. (C) 2007 Elsevier Ltd. All fights reserved.

Cohabitation with a sick cage mate: Effects on ascitic form of Ehrlich tumor growth and macrophage activity

The present study was designed to evaluate the effects of mice cohabitation with a sick conspecific cage mate on peritoneal macrophage activity and on resistance to Ehrlich tumor growth. Female mice housed in pairs were divided into control and experimental groups. One mouse of each control pair was inoculated with NaCl (0.1 ml/10 g) intraperitoneally and the other, called `companion of healthy partner` (CHP), was kept undisturbed. One animal of each experimental pair of mice was inoculated with 5.0 x 10(6) Ehrlich tumor cells intraperitoneally and the other, the subject of this study, was called `companion of sick partner` (CSP). Peritoneal macrophages were removed from CSP and CHP mice to analyze resident macrophage activity (experiment 1), macrophage activity after Mycobacterium bovis (experiment 2) or Ehrlich tumor cells (experiment 3) in vivo inoculations. The resistance of CSP and CHP mice to Ehrlich tumor growth was also analyzed (experiment 4). Differences between groups were not found on resident macrophage activity. However, Onco-BCG- and Ehrlich tumor-activated macrophages from CSP mice presented a decreased intensity and percentage of phagocytosis and an increased respiratory burst in the presence of Staphylococcus aureus stimulation in vitro. CSP animals at the same time displayed a decreased resistance to Ehrlich tumor growth. These data were discussed in light of a possible psychological stress effect imposed by the housing condition on mice`s peritoneal macrophage activity and, as a consequence, on their resistance to Ehrlich tumor growth. Copyright (c) 2008 S. Karger AG, Basel.

Microleakage into and from Two-Stage Implants: An In Vitro Comparative Study

Purpose: Gaps between an abutment and a dental implant are unavoidable, and microleakage may occur, leading to problems such as malodor and peri-implantitis. The aim of the present in vitro study was to investigate leakage of Staphylococcus aureus through the implant/abutment interface by the method of bacterial culture, and to compare the leakage rates of two different types of implant-abutment connections. Materials and Methods: Twenty Morse taper implants with abutments were divided into two groups: group A, which were evaluated for microleakage into the inner part of the implants, and group B, which were evaluated for microleakage from the inner part of the implants. Twenty internal-hexagon implants with abutments were also divided into two groups: group C, which were evaluated for microleakage into the inner part of the implants, and group D, which were evaluated for microleakage from the inner part of the implants. For the evaluation of leakage from the implants, the assemblies had the inner parts inoculated with S aureus, and each assembly was incubated in sterile brain heart infusion broth for 1 week. For assessment of leakage into the implants, each assembly was submerged in 4 mL S aureus culture in tubes and incubated for 2 weeks. The microleakage of the two implant connections was compared. Results: Microbial leakage occurred in all groups, and there was no statistically significant difference between groups A and C or between groups B and D. Conclusions: In vitro, S aureus leakage through the implant/abutment interface occurred with both Morse taper and internal-hexagon implants. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:56-62

Post-antibiotic growth suppression of linezolid against Gram-positive bacteria

The in vitro post-antibiotic effects (PAEs) of eight different concentrations of linezolid against Gram-positive cocci were investigated and the results analysed using the sigmoid E-max model for mathematically modelling the PAE. Mean maximal linezolid PAEs against strains of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium and Streptococcus pneumoniae were 2.2, 1.8, 2.8, 2.0 and 3.0 h, respectively. Resistance to methicillin (for the staphylococci), vancomycin (for the enterococci) and penicillin (for the pneumococci) had no effect on the duration of the PAE. Results of PAE testing support twice-daily dosing of linezolid in humans.

A survey of bacterial diversity in ticks, lice and fleas from Australia

We isolated bacteria from ticks, lice and fleas. Partial small subunit rRNA sequences were obtained for each isolate and the closest matches in the FastA database were determined. These bacteria were mostly Gram-positive (Firmicutes), although representatives from the Proteobacteria (alpha, beta, gamma subdivisions) and CFB group were also isolated. Most of the isolates we found were from genera that were present in most of the ectoparasites studied, but a few genera were restricted to one species of ectoparasite. The most commonly isolated genera were Stenotrophomonas, Staphylococcus, Pseudomonas, Acinetobacter and Bacillus. Species of Bacillus and Proteus, which have biopesticide potential, were found in some of these ectoparasites. Overall, the communities of bacteria were similar to those found in other studies of parasitic arthropods.

Efeito da adição de óleo essencial de pimenta rosa (Schinus terebinthifolius Raddi) microencapsulado em queijo minas frescal

A pimenta rosa é o fruto da aroeira (Schinus terebinthifolius Raddi), uma planta nativa do Brasil. A extração e caracterização do seu óleo essencial viabiliza a sua utilização industrial. Este trabalho teve como objetivo avaliar o efeito da adição de óleo essencial de pimenta rosa em queijo Minas Frescal . Foram identificados 95,36% dos constituintes do óleo essencial, sendo o δ-careno-3 o componente majoritário. O óleo essencial de pimenta rosa apresentou características antioxidante (2,53 ± 0,28 μmols de trolox por mL de óleo) e antimicrobiana para o desenvolvimento de Staphylococcus aureus. Foi observado, no teste de difusão em ágar, a formação do halo de inibição de 1,35 ± 0,32 cm, e na análise de concentração inibitória mínima, o valor de 3,13%. O óleo essencial é sensível à degradação no meio em que se encontra, portanto necessita de processos tecnológicos para assegurar sua ação. Para proteger o óleo essencial foi realizada a microencapsulação por meio da secagem por spray dryer. O óleo foi microencapsulado utilizando três formulações com diferentes concentrações de goma arábica, maltodextrina e amido modificado como papel de parede. As microcápsulas obtidas foram analisadas quanto às suas características físico-químicas e morfológicas e foi selecionada uma formulação para posterior adição ao queijo Minas Frescal. A formulação com 5% de amido modificado, 10% de maltodextrina e 5% de goma arábica foi selecionada. Para definição da concentração de óleo essencial que foi utilzada no queijo, foi realizado teste sensorial de ordenação-preferência, visto que a concentração inibitória mínima (3,13%) era muito alta para ser utilizada nos queijos. A formulação com 0,01% de óleo essencial foi a preferida e utilizada para fabricação dos queijos. Foram elaborados e caracterizados os queijos controle (CO) e o queijo com adição das microcápsulas de óleo essencial de pimenta rosa (OEPR). Em relação às características físico-químicas, os queijos CO e OEPR só apresentaram diferença significativa em relação à umidade (P= 0,0021; α=0,05). O queijo OEPR teve boa aceitação sensorial com média de aceitação global de 7,6 ± 0,97, não apresentando diferença significativa, ao nível de 95% de confiança, do queijo controle (7,6 ± 1,18). Para avaliar a capacidade do óleo essencial de pimenta rosa microencapsulado em inibir o desenvolvimento de S. aureus, foram inoculadas nos queijos CO e OEPR concentrações iniciais de 106 UFG.g. Os queijos foram armazenados em BOD à temperatura de 4 ± 1 °C até realização das análises, que ocorreram nos tempos: 0, 3, 6, 9, 12, 15 e 30 dias. Houve redução de 1,53 ciclos Log no queijo adicionado de óleo essencial de pimenta rosa. De acordo com os resultados obtidos, conclui-se que o óleo essencial de pimenta rosa é uma alternativa viável de utilização em queijo Minas Frescal , devido ao seu potencial antioxidante, antimicrobiano e à sua aceitação sensorial.

Microbiologia dos seios maxilar e etmoidal em pacientes com rinossinusite crônica submetidos à cirurgia funcional endoscópica dos seios paranasais

Estudos da microbiologia da rinossinusite crônica mostram a presença de microorganismos aeróbicos, anaeróbicos, fungos e vírus e sua incidência varia de acordo com cada estudo. Estes estudos nos guiam para a escolha do antimicrobiano mais adequado para eliminar o processo infeccioso, ajudando a restaurar a mucosa nasossinusal. FORMA DE ESTUDO: Clínico prospectivo. OBJETIVO: O objetivo deste trabalho foi estudar a microbiologia dos seios maxilar e/ou etmoidal de pacientes com rinossinusite crônica e com indicação de cirurgia funcional endoscópica dos seios paranasais. MATERIAIS E MÉTODOS: Durante a cirurgia coletamos, em 41 pacientes, secreção e/ou fragmento de mucosa dos seios maxilar e/ou etmoidal para realização de bacterioscopia, pesquisa direta de fungos, cultura para microorganismos aeróbios, anaeróbios e fungos. RESULTADOS: Identificou-se a presença de microorganismos aeróbios em 21 pacientes (51,2%), anaeróbios em 16 (39%) e fungos em 1 (2,4%). Na população estudada, apenas em 12 (29,2%) o microorganismo isolado foi considerado patogênico quando analisado junto à contagem semiquantitativa de leucócitos. O Staphylococcus coagulase-negativo e o Staphylococcus aureus foram os microorganismos mais freqüentes, em 5 (12,1%) e em 4 pacientes (9,75%) respectivamente. CONCLUSÃO: Este estudo revela que o Staphylococcus coagulase-negative e o Staphylococcus aureus foram os microorganismos mais freqüentes isolados nos pacientes com rinossinusite crônica.

Bacteriologia do aspirado do meato médio em pacientes com fibrose cística

A combinação de fatores como viscosidade das secreções dos seios paranasais, diminuição da drenagem sinusal e comprometimento do transporte mucociliar podem ser responsáveis pela criação de um ambiente propício e adequado para a colonização de bactérias nos seios paranasais de pacientes com fibrose cística. OBJETIVO: Analisar a bacteriologia do aspirado do meato médio de pacientes portadores de fibrose cística. MATERIAL E MÉTODO: Através de um estudo prospectivo de delineamento transversal, avaliou-se uma amostra composta de 23 pacientes, avaliados durante 2 anos. Realizaram-se relações entre a cultura do meato médio e a avaliação radiológica do seio maxilar e a avaliação clínica. Secundariamente, estudou-se a relação da bacteriologia do aspirado do meato médio e a do escarro. RESULTADOS: No total foram realizadas 42 aspirações do meato médio. Em 17 (73,91%) dos 23 pacientes, as culturas foram negativas e, em 6 (26,08%), positivas. Das 42 aspirações, 31 (73,8%) foram negativas e 11 (26,2%), positivas. A presença de Pseudomonas aeruginosa foi observada em 18,18% das culturas positivas e o Staphylococcus aerus em 27,28%. CONCLUSÃO: A maioria das culturas do aspirado do meato médio de pacientes com fibrose cística foi negativa.