Isolamento de Salmonella enterica em gambás (Didelphis aurita e Didelphis albiventris) do Estado de São Paulo, Brasil

No Brasil, não há relato de estudos de Salmonella em gambás, sendo assim, este trabalho tem por objetivo determinar a frequência de isolamento de Salmonella enterica em gambás (D. aurita e D. albiventris) no Estado de São Paulo. No período de janeiro de 2005 a dezembro de 2006, foram necropsiados 106 D. aurita e 40 D. albiventris e colhidos fragmentos de intestinos delgado, grosso e suabe da cloaca. As amostras foram plaqueadas diretamente em ágar Mac Conkey, paralelamente suspendidas nos caldos Rappaport-Vassiliadis e Tetrationato e posteriormente plaqueados em ágar XLT4. As colônias sugestivas de Salmonella foram confirmadas através de provas bioquímicas e sorotipagem. Encontrou-se Salmonella enterica em 17,0% (18/106) dos D. aurita. Destes, 50% apresentaram positividade no intestino delgado (ID), 88,9% no intestino grosso (IG) e 66,7% na cloaca. Da espécie S. enterica, as subespécies encontradas foram: diarizonae (11,1%) houtenae e enterica (5,5% cada um); enquanto da subespécie S. enterica enterica os sorotipos foram Newport (83,3%), Typhimurium e Cerro (5,5% cada um). Nos D. albiventris, 17,5% (7/40) eram positivos, sendo que se encontraram 42,8% no ID, 85,7% no IG e 71,4% na cloaca. O sorotipo mais prevalente também foi Newport (71,4%), seguido por Typhimurium, Bareilly e Thompson (14,3% cada um). Através dos resultados obtidos neste estudo pode-se comprovar a presença de Salmonella enterica no trato intestinal de gambás no Brasil.

Modulatory effect of Byrsonima basiloba extracts on the mutagenicity of certain direct and indirect-acting mutagens in Salmonella typhimurium assays

Byrsonima basiloba A. Juss. species is a native arboreal type from the Brazilian ""cerrado"" (tropical American savanna), and the local population uses it to treat diseases, such as diarrhea and gastric ulcer. It belongs to the Malpighiaceae family, and it is commonly known as ""murici."" Considering the popular use of B. basiloba derivatives and the lack of pharmacological potential studies regarding this vegetal species, the mutagenic and antimutagenic effect of methanol (MeOH) and chloroform extracts were evaluated by the Ames test, using strains TA97a, TA98, TA100, and TA102 of Salmonella typhimurium. No mutagenic activity was observed in any of the extracts. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine, sodium azide, mitomycin C, aflatoxin B(1), benzo[a] pyrene, and hydrogen peroxide. Both the extracts evaluated showed antimutagenic activity, but the highest value of inhibition level (89%) was obtained with the MeOH extract and strain TA100 in the presence of aflatoxin B(1). Phytochemical analysis of the extracts revealed the presence of n-alkanes, lupeol, ursolic and oleanolic acid, (+)-catechin, quercetin- 3-O-alpha-L-arabinopyranoside, gallic acid, methyl gallate, amentoflavone, quercetin, quercetin-3-O-(2 ''-O-galloyl)-beta-D-galactopyranoside, and quercetin-3-O-(2 ''-O-galloyl)-alpha-L-arabinopyranoside.

Spatial patterns of malaria in a land reform colonization project, Juruena municipality, Mato Grosso, Brazil

Background: In Brazil, 99% of malaria cases are concentrated in the Amazon, and malaria's spatial distribution is commonly associated with socio-environmental conditions on a fine landscape scale. In this study, the spatial patterns of malaria and its determinants in a rural settlement of the Brazilian agricultural reform programme called ""Vale do Amanhecer"" in the northern Mato Grosso state were analysed. Methods: In a fine-scaled, exploratory ecological study, geocoded notification forms corresponding to malaria cases from 2005 were compared with spectral indices, such as the Normalized Difference Vegetation Index (NDVI) and the third component of the Tasseled Cap Transformation (TC_3) and thematic layers, derived from the visual interpretation of multispectral TM-Landsat 5 imagery and the application of GIS distance operators. Results: Of a total of 336 malaria cases, 102 (30.36%) were caused by Plasmodium falciparum and 174 (51.79%) by Plasmodium vivax. Of all the cases, 37.6% (133 cases) were from residents of a unique road. In total, 276 cases were reported for the southern part of the settlement, where the population density is higher, with notification rates higher than 10 cases per household. The local landscape mostly consists of open areas (38.79 km(2)). Training forest occupied 27.34 km(2) and midsize vegetation 7.01 km(2). Most domiciles with more than five notified malaria cases were located near areas with high NDVI values. Most domiciles (41.78%) and malaria cases (44.94%) were concentrated in areas with intermediate values of the TC_3, a spectral index representing surface and vegetation humidity. Conclusions: Environmental factors and their alteration are associated with the occurrence and spatial distribution of malaria cases in rural settlements.

Vaccination of Mice with Salmonella Expressing VapA: Mucosal and Systemic Th1 Responses Provide Protection against Rhodococcus equi Infection

Conventional vaccines to prevent the pneumonia caused by Rhodococcus equi have not been successful. We have recently demonstrated that immunization with Salmonella enterica Typhimurium expressing the VapA antigen protects mice against R. equi infection. We now report that oral vaccination of mice with this recombinant strain results in high and persistent fecal levels of antigen-specific IgA, and specific proliferation of the spleen cells of immunized mice in response to the in vitro stimulation with R. equi antigen. After in vitro stimulation, spleen cells of immunized mice produce high levels of Th1 cytokines and show a prominent mRNA expression of the Th1 transcription factor T-bet, in detriment of the Th2 transcription factor GATA-3. Following R. equi challenge, a high H(2)O(2), NO, IL-12, and IFN-gamma content is detected in the organs of immunized mice. On the other hand, TNF-alpha and IL-4 levels are markedly lower in the organs of vaccinated mice, compared with the non-vaccinated ones. The IL-10 content and the mRNA transcription level of TGF-beta are also higher in the organs of immunized mice. A greater incidence of CD4(+) and CD8(+) T cells and B lymphocytes is verified in vaccinated mice. However, there is no difference between vaccinated and non-vaccinated mice in terms of the frequency of CD4(+)CD25(+)Foxp3(+) T cells. Finally, we show that the vaccination confers a long-term protection against R. equi infection. Altogether, these data indicate that the oral vaccination of mice with S. enterica Typhimurium expressing VapA induces specific and long-lasting humoral and cellular responses against the pathogen, which are appropriately regulated and allow tissue integrity after challenge.

Study of Salmonella typhimurium mutagenicity assay of (E)-piplartine by the Ames test

Phytochemical studies carried out with Piperaceae species have shown great diversity of secondary metabolites among which are several displayed considerable biological activities. The species Piper tuberculatum has been intensively investigated and a series of amides have been described. For instance, (E)-piplartine showed significant cytotoxic activity against tumor cell lines, especially human leukemia cell lines; antifungal activity against Cladosporium species; trypanocidal activity and others. Considering the popular use of P. tuberculatum and the lack of pharmacological studies regarding this plant species, the mutagenic and antimutagenic effect of (E)-piplartine was evaluated by the Ames test, using the strains TA97a, TA98, TA100 and TA102 of Salmonella typhimurium. No mutagenic activity was observed for this compound.

Colonization and extinction of land planarians (Platyhelminthes, Tricladida) in a Brazilian Atlantic Forest regrowth remnant

Long-term assessments of species assemblages are valuable tools for detecting species ecological preferences and their dispersal tracks, as well as for assessing the possible effects of alien species on native communities. Here we report a 50-year-long study on population dynamics of the four species of land flatworms (Platyhelminthes, Tricladida, Terricola) that have colonized or become extinct in a 70-year-old Atlantic Forest regrowth remnant through the period 1955-2006. On the one hand, the two initially most abundant species, which are native to the study site, Notogynaphallia ernesti and Geoplana multicolor have declined over decades and at present do not exist in the forest remnant. The extinction of these species is most likely related with their preference for open vegetation areas, which presently do not exist in the forest remnant. On the other hand, the neotropical Geoplaninae 1 and the exotic Endeavouria septemlineata were detected in the forest only very recently. The long-term study allowed us to conclude that Geoplaninae 1 was introduced into the study area, although it is only known from the study site. Endeavouria septemlineata, an active predator of the exotic giant African snail, is originally known from Hawaii. This land flatworm species was observed repeatedly in Brazilian anthropogenic areas, and this is the first report of the species in relatively well preserved native forest, which may be evidence of an ongoing adaptive process. Monitoring of its geographic spread and its ecological role would be a good practice for preventing potential damaging effects, since it also feeds on native mollusk fauna, as we observed in lab conditions.

Endophytic Colonization of Potato (Solanum tuberosum L.) by a Novel Competent Bacterial Endophyte, Pseudomonas putida Strain P9, and Its Effect on Associated Bacterial Communities

Pseudomonas putida strain P9 is a novel competent endophyte from potato. P9 causes cultivar-dependent suppression of Phytophthora infestans. Colonization of the rhizoplane and endosphere of potato plants by P9 and its rifampin-resistant derivative P9R was studied. The purposes of this work were to follow the fate of P9 inside growing potato plants and to establish its effect on associated microbial communities. The effects of P9 and P9R inoculation were studied in two separate experiments. The roots of transplants of three different cultivars of potato were dipped in suspensions of P9 or P9R cells, and the plants were planted in soil. The fate of both strains was followed by examining colony growth and by performing PCR-denaturing gradient gel electrophoresis (PCR-DGGE). Colonies of both strains were recovered from rhizoplane and endosphere samples of all three cultivars at two growth stages. A conspicuous band, representing P9 and P9R, was found in all Pseudomonas PCR-DGGE fingerprints for treated plants. The numbers of P9R CFU and the P9R-specific band intensities for the different replicate samples were positively correlated, as determined by linear regression analysis. The effects of plant growth stage, genotype, and the presence of P9R on associated microbial communities were examined by multivariate and unweighted-pair group method with arithmetic mean cluster analyses of PCR-DGGE fingerprints. The presence of strain P9R had an effect on bacterial groups identified as Pseudomonas azotoformans, Pseudomonas veronii, and Pseudomonas syringae. In conclusion, strain P9 is an avid colonizer of potato plants, competing with microbial populations indigenous to the potato phytosphere. Bacterization with a biocontrol agent has an important and previously unexplored effect on plant-associated communities.

Fate of Surface-Inoculated Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium on Kippered Beef during Extended Storage at Refrigeration and Abusive Temperatures

The behavior of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium on kippered beef was evaluated. Individual pieces of the product were separately inoculated on the top and bottom surfaces with each three- to six-strain pathogen cocktail at ca. 6.0 log CFU per piece and stored at 4, 10, 21, or 30 degrees C for up to 28 days in each of two trials. When kippered beef was inoculated with E coli O157:H7, Salmonella Typhimurium, or L. monocytogenes and stored at 4, 10, 2 1, or 30 degrees C for up to 28 days, pathogen numbers decreased ca. 0.4 to 0.9, 1.0 to 1.8, 3.0 to >= 5.25, and >= 5.0 to 5.25 log CFU per piece, respectively. Average D-values for E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes stored at 4 to 30 degrees C for 28 days were ca. 41 to 4.6, 40.8 to 5.3, and 29.5 to 4.3 days, respectively. As expected, the higher the storage temperature, the greater the level and rate of inactivation for all three pathogens. These data establish that kippered beef does not provide an environment conducive to proliferation of these pathogens.

Diversity of endophytic bacteria from Eucalyptus species seeds and colonization of seedlings by Pantoea agglomerans

The diversity and beneficial characteristics of endophytic microorganisms have been studied in several host plants. However, information regal-ding naturally, occurring seed-associated endophytes and vertical transmission among different life-history stages of hosts is limited. Endophytic bacteria were isolated from seeds and seedlings of 10 Eucalyptus species and two hybrids. The results showed that endophytic bacteria, Such as Bacillus, Enterococcus, Paenibacillus and Methylobacterium, are vertically transferred from seeds to seedlings. In addition, the endophytic bacterium Pantoea agglomerans was tagged with the gfp gene, inoculated into seeds and further reisolated from seedlings. These results suggested it novel approach to change the profile of the plants, where the bacterium is a delivery vehicle for desired traits. This is the first report of an endophytic bacterial community residing in Eucalyptus seeds and the transmission of these bacteria from seeds to seedlings. The bacterial species reported ill this work have been described as providing benefits to host plants. Therefore, we Suggest that endophytic bacteria can be transmitted vertically from seeds to seedlings, assuring the support of the bacterial community in the host plant.

Listeria monocytogenes and Salmonella spp. in raw milk produced in Brazil: Occurrence and interference of indigenous microbiota in their isolation and development

This study aimed to verify the occurrence of Listeria monocytogenes and Salmonella spp. in raw milk produced in Brazil. On account of the poor microbiological quality of this product, possible interference from the indigenous microbiota in these pathogens was also evaluated. Two-hundred and ten raw milk samples were collected in four important milk-producing areas in Brazil, tested for L. monocytogenes and Salmonella spp. presence, and for enumeration of indicator microorganisms: mesophilic aerobes, total coliforms and Escherichia coli. The interference of the indigenous microbiota in the isolation procedures was also tested, as well the frequency of naturally occurring raw milk strains with antagonistic activity against both pathogens. The pathogens were not isolated in any raw milk sample, but poor microbiological quality was confirmed by the high levels of indicator microorganisms. When present at high levels, the indigenous microbiota generated an evident interference in the methodologies of L. monocytogenes and Salmonella spp. isolation, mainly when the pathogens appeared at low levels. Three-hundred and sixty raw milk strains were tested for antagonistic activity against both pathogens, and 91 (25.3%) showed inhibitory activity against L. monocytogenes and 33 (9.2%) against Salmonella spp. The majority of the antagonistic strains were identified as Lactic Acid Bacteria species, mainly Lactococcus lactis subsp. lactis and Enterococcus faecium, known by antimicrobial substance production.

Radioresistance of Salmonella species and Listeria monocytogenes on minimally processed arugula (Eruca sativa Mill.): Effect of irradiation on flavonoid content and acceptability of irradiated produce

This work studied the radiation resistance of Listeria monocytogenes and Salmonella species and the effect of irradiation on leaf flavonoid content and sensory acceptability of minimally processed arugula. Immersion in ozone-treated water reduced the analyzed microorganisms by 1 log. L. monocytogenes and Salmonella were not isolated from samples. Samples of this vegetable were inoculated with a cocktail of Salmonella spp. and L. monocytogenes and exposed to gamma irradiation. D-10 values for Salmonella ranged from 0.16 to 0.19 kGy and for L. monocytogenes from 0.37 to 0.48 kGy. Kaempferol glycoside levels were 4 and ca. 3 times higher in samples exposed to 1 and 2 kGy, respectively, than in control samples. An increase in quercetin glycoside was also observed mainly in samples exposed to 1 kGy. In sensory evaluation, arugula had good acceptability, even after exposure to 2 and 4 kGy. These results indicate that irradiation has potential as a practical processing step to improve the safety of arugula.

The introduction of the Salmonella/microsome mutagenicity assay in a groundwater monitoring program

The objective of this study was to verify the possible inclusion of the Salmonella/microsome mutagenicity assay in a groundwater monitoring program as a complementary assay to assess water quality. Groundwater samples belonging to seven wells from different types of aquifers were analyzed. Three different methods for sample preparation were used: membrane filtration; liquid-liquid and XAD-4 extraction. The filtered samples were tested using TA98, TA100, YG1041 and YG1042 and the water extracts only with TA98 and TA100. No mutagenic activity was observed in any of the 16 filtered samples tested. Out of the 10 samples analyzed using XAD-4 extraction, five showed mutagenic activity with potency ranging from 130 to 1500 revertants/L. Concerning the liquid-liquid extraction, from the 11 samples analyzed, 3 showed mutagenicity. The XAD-4 extraction was the most suitable sample preparation. TA98 without S9 was found to be the most sensitive testing condition. The wells presenting water samples with mutagenic activity belonged to unconfined aquifers, which are more vulnerable to contamination. The data suggest that Salmonella/microsome assay can be used as an efficient screening tool to monitor groundwater for mutagenic activity. (C) 2009 Elsevier B.V. All rights reserved.

Comparison of the Salmonella/Microsome Microsuspension Assay with the new Microplate Fluctuation Protocol for Testing the Mutagenicity of Environmental Samples

The objective of this study was to compare the responses of the Salmonella/microsome microsuspension assay with the new microplate fluctuation protocol (MPF) for the evaluation of the mutagenic activity of environmental samples. Organic extracts of total particulate atmospheric air samples, surface waters, and effluents were tested in dose-response experiments. The assays were performed with strain TA98 in the absence and presence of S9 mix. Both protocols produced similar results, despite the fact that the maximum score of the MPF is limited to 48 wells, whereas in the regular plate assay it is possible to count up to 1,500 colonies using an automatic counter. Similar sensitivities based on the lowest dose that resulted in a positive response were obtained for both assays. The MPF procedure is less laborious (e.g., all-liquid format, use of multi-channel pipettors) and allows for automation of the pipetting and dispensing steps, thus, reducing time of the analysis which is particularly important in environmental quality monitoring programs or in effect-directed analysis. The results show that the MPF procedure is a promising tool to test environmental samples for mutagenic activity. Environ. Mol. Mutagen. 51:31-38, 2010. (C) 2009 Wiley-Liss, Inc.

OCCURRENCE AND ANTIBIOTIC SENSIBILITY PROFILE OF Salmonella spp. ISOLATED FROM PORK MEAT CUTS COMMERCIALIZED IN THE OPEN MARKETS IN PELOTAS, RS (BRAZIL)

The objective of the present study was to evaluate the occurrence of Salmonella spp. in 15 samples of pork meat cuts (T-bone, shank, sausage and ribs) commercialized in open markets of Pelotas (RS, Brazil) and verify the prevalent serovars, and test the isolates profile of sensitivity to several antibiotics of importance in medicine (nalidixic acid, ampicillin, aztreonam, kanamycin, carbenicillin, cephalothin, cefoxitin, ceftriaxone, ciprofloxacin, chloramphenicol, gentamicin, sulfonamide, tetracycline and trimetoprina). Twelve samples (80%) were contaminated by Salmonella enterica, serovars Infantis, Derby, Panama and Typhimurium. All isolates were susceptible to trimetoprin, aztreonam, ciprofloxacin, ceftriaxone and cefoxitin. For the other antibiotics, the pattern of sensitivity varied as serovar. In addition, 39.1% of isolates showed up to be multiresistant.

Colonization and risk factors for Brachyspira aalborgi and Brachyspira pilosicoli in humans and dogs on tea estates in Assam, India

The prevalence of colonization with the anaerobic intestinal spirochaetes Brachyspira aalborgi and Brachyspira pilosicoli was investigated in humans (n = 316) and dogs (n = 101) living on three tea estates in Assam, India. Colonization was detected using PCR on DNA from faeces. Nineteen (6%) human faecal samples contained B. aalborgi DNA, 80 (25.3%) contained B. pilosicoli DNA, and 10 (3.2%) contained DNA from both species. One canine sample contained DNA from B. pilosicoli. Significant factors for B. aalborgi colonization in logistic regression were: infection of family members with B. aalborgi (P < 0.001), being a resident of Balipara (P = 0.03), and use of water treatment (P = 0.03). For B. pilosicoli, significant factors were: other family members being positive for B. pilosicoli (P < 0.001), water obtained from a well (P = 0.006), water treatment (P = 0.03), and not having visited a doctor in the previous 12 months (P = 0.03).