Investigating luxS gene expression in lactobacilli along lab-scale cocoa fermentations.

Previous metagenomic analyses have suggested that lactobacilli present potential for Quorum Sensing (QS) in cocoa fermentation, and in the present research, laboratory scale fermentations were carried out to monitor the expression of luxS, a universal marker of QS. For that, 96 h-fermentations were studied, as follows: F0 (non inoculated control), F1 (inoculated with yeasts, lactic acid bacteria, and acetic acid bacteria), F2 (inoculated with yeasts and acetic acid bacteria), F3 (inoculated with yeasts only). The parameters evaluated were: plate counting, quantification of key enzymes and analysis of volatile organic compounds associated with key sensory descriptors, using headspace gas chromatography-mass spectrometry (GC-MS). Furthermore, QS was estimated by the quantification of the expression of luxS genes by Reverse Transcriptase Real-Time PCR. The results demonstrated that microbial succession occurred in pilot scale fermentations, but no statistical differences for microbial enumeration and α-diversity index were observed among experiments and control. Moreover, it was not possible to make conclusive correlations of enzymatic profile and fermenting microbiota, likely due to the intrinsic activity of plant hydrolases. Regarding to the expression of luxS genes, in Lactiplantibacillus plantarum they were active along the fermentation, but for Limosilactobacillus fermentum, luxS was expressed only at early and middle phases. Correlation analysis of luxS expression and production of volatile metabolites evidenced a possible negative association of Lp. Plantarum with fermentation quality. In conclusion, these data corroborate former shotgun metagenomic analysis by demonstrating the expression of luxS by lactobacilli in pilot scale cocoa fermentation and evidence Lp. Plantarum is the main lactic acid bacteria related to its expression.

Simplified magnetic anchor-guided endoscopic submucosal dissection: an ex vivo porcine model.

INTRODUCTION AND AIMS: Endoscopic submucosal dissection (ESD) is a well-established treatment for superficial gastrointestinal tumors and enables en bloc resection. Adequate tissue tension is important for safe and effective dissection. Simplified magnetic anchor-guided ESD (MAG-ESD) with a neodymium magnet has potential benefits, compared with other current traction methods. We evaluated the feasibility of simplified MAG-ESD in an ex vivo porcine model. MATERIALS AND METHODS: An experimental study was conducted, utilizing the standard ESD technique. An external magnet and an internal magnet, both neodymium magnets, were used for the magnetic anchoring. The internal magnet was attached to an arm of a hemoclip with a 2-0 silk suture. After the incision, the clip with the internal magnet was placed at the edge of the lesion. The external magnet was maneuvered around the surface to apply adequate tension. RESULTS: A total of 15 en bloc ESDs (5 with no magnetic anchoring and 10 with magnetic anchoring) were carried out. Traction and dissection were feasible in all cases and the procedures were completed in fewer than 90min. Lesion size ranged from 15 to 50mm (mean 30mm). Two cases in the group with magnetic anchoring presented with punctate perforation (13.3%). CONCLUSIONS: Our study demonstrated the feasibility of simplified MAG-ESD and en bloc resection in an ex vivo porcine model.

Simplified magnetic anchor-guided endoscopic submucosal dissection: an ex vivo porcine model. / Disección endoscópica de la submucosa asistida con anclaje magnético simplificado: modelo porcino ex vivo.

INTRODUCTION AND AIMS: Endoscopic submucosal dissection (ESD) is a well-established treatment for superficial gastrointestinal tumors and enables en bloc resection. Adequate tissue tension is important for safe and effective dissection. Simplified magnetic anchor-guided ESD (MAG-ESD) with a neodymium magnet has potential benefits, compared with other current traction methods. We evaluated the feasibility of simplified MAG-ESD in an ex vivo porcine model. MATERIALS AND METHODS: An experimental study was conducted, utilizing the standard ESD technique. An external magnet and an internal magnet, both neodymium magnets, were used for the magnetic anchoring. The internal magnet was attached to an arm of a hemoclip with a 2-0 silk suture. After the incision, the clip with the internal magnet was placed at the edge of the lesion. The external magnet was maneuvered around the surface to apply adequate tension. RESULTS: A total of 15 en bloc ESDs (five with no magnetic anchoring and 10 with magnetic anchoring) were carried out. Traction and dissection were feasible in all cases and the procedures were completed in fewer than 90 min. Lesion size ranged from 15 to 50 mm (mean 30 mm). Two cases in the group with magnetic anchoring presented with punctate perforation (13.3%). CONCLUSIONS: Our study demonstrated the feasibility of simplified MAG-ESD and en bloc resection in an ex vivo porcine model.

Evidence for the occurrence of LDL receptors in extracts of schistosomula of Schistosoma mansoni.

Schistosomula of Schistosoma mansoni were shown to contain proteins on their surface membranes which bind iodinated human low density lipoproteins (125I-LDL). Treatment of the parasites with trypsin decreased the binding in comparison with untreated controls. Membrane-bound, acetone-insoluble proteins were extracted from the schistosomula with Triton X-100 and the extract in liposome form was incubated with 125I-LDL at room temperature. After incubation a complex was formed between the proteins present in the extract and 125I-LDL, as shown by a filter binding assay. 125I-LDL binding to filters was proportional to the amount of protein in the extract; it was inhibited by unlabelled LDL and VLDL and by EDTA. Binding of 125I-LDL to proteins present in the liposome suspension containing the Triton X-100 extract followed saturation kinetics, indicating the occurrence of receptors for lipoproteins in the extract.

Characterisation of Fasciola hepatica cytochrome c peroxidase as an enzyme with potential antioxidant activity in vitro.

Cytochrome c peroxidase oxidises hydrogen peroxide using cytochrome c as the electron donor. This enzyme is found in yeast and bacteria and has been also described in the trematodes Fasciola hepatica and Schistosoma mansoni. Using partially purified cytochrome c peroxidase samples from Fasciola hepatica we evaluated its role as an antioxidant enzyme via the investigation of its ability to protect against oxidative damage to deoxyribose in vitro. A system containing FeIII-EDTA plus ascorbate was used to generate reactive oxygen species superoxide radical, H2O2 as well as the hydroxyl radical. Fasciola hepatica cytochrome c peroxidase effectively protected deoxyribose against oxidative damage in the presence of its substrate cytochrome c. This protection was proportional to the amount of enzyme added and occurred only in the presence of cytochrome c. Due to the low specific activity of the final partially purified sample the effects of ascorbate and calcium chloride on cytochrome c peroxidase were investigated. The activity of the partially purified enzyme was found to increase between 10 and 37% upon reduction with ascorbate. However, incubation of the partially purified enzyme with 1 mM calcium chloride did not have any effect on enzyme activity. Our results showed that Fasciola hepatica CcP can protect deoxyribose from oxidative damage in vitro by blocking the formation of the highly toxic hydroxyl radical (.OH). We suggest that the capacity of CcP to inhibit .OH-formation, by efficiently removing H2O2 from the in vitro oxidative system, may extend the biological role of CcP in response to oxidative stress in Fasciola hepatica.

Investigations of the vertebrate hosts of eastern equine encephalitis during an epizootic in Michigan, 1980.

A study was undertaken to investigate an increase in reported cases of clinical encephalitis due to eastern equine encephalitis (EEE) virus in horses and to determine the natural vertebrate hosts of that virus. Horses, birds, and small mammals were sampled at sites in a contiguous area in St. Joseph and Kalamazoo counties, Michigan, from 25 August to 5 September 1980. Serum samples from four horses acutely ill with encephalitis and 16 of 39 pasture mates of ill horses had neutralizing (N) antibody against EEE virus (46.5%); no viruses were isolated from these 43 sera. None of 24 draft horses from a site in St. Joseph County 12 km southeast of the affected sites had EEE antibody. A strain of Cache Valley virus was isolated from the blood of one of the 24 draft horses. No viruses were isolated, and no antibodies to EEE virus were detected in 28 blood samples from small mammals captured at sites where equine cases of encephalitis were occurring. Six strains of EEE virus, five of Highlands J virus, and one of Flanders virus were isolated from the blood of 401 wild birds belonging to 42 species captured at eight sites in both counties. A total of 29.9% of the wild birds had EEE antibody. Five species of domestic birds, mostly chickens and ring-necked pheasants, were sampled in both counties. Six strains of EEE virus were isolated from 152 ring-necked pheasants; these included three isolates from the brains of dead birds. About 13% of 51 pheasants tested from two small flocks in backyard pens in Kalamazoo County and 9% of 103 pheasants tested from a large game farm in St. Joseph County had antibody to EEE virus. The source of the EEE virus and the factors responsible for this epizootic are unknown; however, the epizootic probably represented an explosive expansion of an enzootic level of virus transmission.

Experimental studies of St. Louis encephalitis virus in vertebrates.

Serologically negative birds and mammals of species, known from other studies to be exposed naturally to St. Louis encephalitis (SLE) virus in Memphis, Tennessee, and other selected species were inoculated experimentally with strains of SLE virus to determine their potential as natural hosts. Mosquitoes (Culex sp.) were allowed to feed on some of the inoculated vertebrate species, held for 14 days, and tested for SLE infection. The cardinals (Richmondena cardinalis), robins (Turdus migratorius), and baby chicks (Gallus gallus) all became viremic; 97% of the bobwhites (Colinus virginianus) and 20% of the Japanese quail (Coturnix coturnix) became viremic. No viremia was detected in raccoons (Procyon lotor), opossums (Didelphis virginiana), or adult cotton rats (Sigmodon hispidus). Only 20% of cottontail rabbits (Sylvilagus audubonii), 50% of wood rats (Neotoma mexicana), and 75% of hamsters (Mesocricetus auratus) but all the young cotton rats and least chipmunks (Eutamias minimus) were susceptible. Robins had the highest titered viremia but were viremic for the shortest period of time. Bobwhites had lower peak viremia titers but for a longer duration. Biologic differences in the response of some vertebrates to different SLE strains were noted. Culex pipiens quinquefasciatus mosquitoes readily became infected after feeding on viremic cardinals. Comparisons of the experimental data with information obtained from field investigations provided a better understanding of the contributions of the various vertebrate species to the transmission and maintenance of SLE virus in nature.

The role of deer as a possible reservoir host of potosi virus, a newly recognized arbovirus in the United States.

Potosi (POT) virus (Bunyaviridae) was isolated from Aedes albopictus, an introduced Asian mosquito species, collected at a used tire yard in Potosi, Missouri (USA), in August and September, 1989. In September, 1990, small animals were trapped at the tire yard and six cattle were sampled at an adjacent farm; in November 1990 and 1991, blood samples were collected with filter paper strips from 364 hunter-killed, white-tailed deer (Odocoileus virginianus) in the region to determine the possible reservoir hosts of the virus. Deer specimens from Arkansas (n = 70), Colorado (n = 29), and Iowa (n = 763) (USA) were also analyzed. Specimens from 33 small vertebrates captured at the tire yard were negative for viruses. Only one eastern chipmunk (Tamias striatus) and none of six cattle had neutralizing (N) antibody against POT virus by the plaque-reduction serum neutralization test in Vero cell culture but 45 (25%) of 178 deer specimens in 1990 and 55 (30%) of 186 in 1991 were antibody positive. The 186 deer sera from 1991 were tested further and 29 (16%) were also N antibody positive to Cache Valley (CV) virus. From the 763 deer specimens tested from Iowa in 1993, 114 (15%) had N antibody to POT virus. Of 70 serum specimens from Arkansas deer in 1990, 33 (47%) had N antibody to POT and 15 (21%) to CV viruses; two (7%) of 29 CV negative serum specimens from Colorado deer in 1981 were serologically positive to POT virus. Three eastern chipmunks were experimentally inoculated with POT virus to determine their reservoir potential; none became viremic but all developed N antibody. Thus we propose that POT virus may be another virus regularly infecting wild deer populations but its impact on the health of these animals is unknown.

Apparent influence of the stage of blood meal digestion on the efficacy of ground applied ULV aerosols for the control of urban Culex mosquitoes. II. Laboratory evidence.

The susceptibility of adult Culex pipiens s.l., Culiseta melanura and Aedes aegypti to insecticide aerosols in wind tunnel exposures varied with time, depending on the stage of blood meal digestion. Greater than 2-fold differences were observed in the concentrations of malathion and synergized resmethrin required to kill test mosquitoes, depending on whether they had been given a blood meal and, if they had, the length of time following the blood meal. The period of lowest susceptibility varied from 24 h after feeding in Ae. aegypti to 72 h in Cs. melanura. The greatest variability occurred during the period when undigested blood was present. Data from tests with a malathion-tolerant strain of Cx. pipiens s.l. suggested little change in susceptibility regardless of blood feeding and the associated weight changes that occur from ingestion of blood.

Apparent influence of the stage of blood meal digestion on the efficacy of ground applied ULV aerosols for the control of urban Culex mosquitoes. I. Field evidence.

The impact of ULV resmethrin on urban Culex mosquitoes was evaluated in 4 field trials by monitoring daily oviposition rate. A well-defined oscillation of effect, with a period corresponding to the duration of the gonotrophic cycle, was observed. We postulate that this oscillation arises from changes in susceptibility following blood feeding and/or behavioral factors. The data indicate that a single treatment with ULV may be inadequate for the effective control of vector mosquitoes.

Apparent influence of the stage of blood meal digestion on the efficacy of ground applied ULV aerosols for the control of urban Culex mosquitoes. III. Results of a computer simulation.

When posttreatment response to ultra low volume (ULV) application of insecticide has been followed for periods approaching a week, a damped oscillation in oviposition rates is observed, probably because females who have been recently blood-fed are more resistant to insecticides than their unfed siblings. We describe a simple model (ULVSIM) that incorporates physiologic changes in insecticide susceptibility and accounts for much of our field data. The model follows 30 cohorts over 30 days following insecticidal treatment. Multiple treatments or short-term residual activity can be evaluated. The model predicts that oviposition will follow a pattern of damped oscillations after an adulticidal treatment. The model gave a good fit to oviposition data obtained in 2 field trials of resmethrin for 7 to 9 days after treatment. It can be used to evaluate the effect of single and multiple treatments on the total female Culex population and on numbers of infected females surviving for different periods following an infective blood meal.

Aedes albopictus and other container-inhabiting mosquitoes in the United States: results of an eight-city survey.

Extensive surveys were conducted in 1987 in Baytown, TX; Lafayette, Shreveport and Baton Rouge, LA; Memphis, TN; Kansas City, MO; Evansville, IN; and Jacksonville, FL. The program objective was to determine the intensity of Aedes albopictus infestations, to evaluate the degree to which Ae. albopictus had spread into residential areas, to document habitat selection and to obtain background information for possible suppression or eradication projects. This report describes the survey methods and presents a preliminary analysis of the data. Larvae, pupae and adult mosquitoes were collected from container habitats in a randomized selection of urban premises as well as at and around sites known to be at high risk for introduction of Ae. albopictus. Adult or larval mosquitoes were collected from 24.4% of 5,728 premises inspected, and there were an average of 3.27 positive containers per positive premise. Several known disease vectors, especially Culex pipiens (s.l.), were frequently found in urban container habitats. The large numbers of specimens collected during the surveys and the detailed information available for each collection make this a useful database for comparison in future studies.

Antibody prevalence of St. Louis encephalitis virus in avian hosts in Los Angeles, California, 1986.

A study was conducted to determine the pattern of St. Louis encephalitis (SLE) virus activity in the avian populations of the Los Angeles metropolitan area in 1986. In total, 679 birds of 42 species were captured at 7 study sites. The overall prevalence of SLE neutralizing (N) antibody of 3% indicated enzootic transmission. Antibody prevalences were higher in birds sampled in the central part of the metropolitan area, which was consistent with other epidemiologic data. The use of specific avian species as sentinels for future surveillance of SLE virus activity was suggested.

Importation of Aedes albopictus and other exotic mosquito species into the United States in used tires from Asia.

Between May 18 and December 4, 1986, 79 seagoing containers and their contents of 22,051 used tires were inspected for adult mosquitoes as well as eggs and larvae. Of the total inspected, 5,507 tires (25%) contained significant amounts of water. No adults or eggs were found. Fifteen tires contained mosquito larvae that were identified as Ae. albopictus, Ae. togoi, Culex pipiens complex, Tripteroides bambusa and Uranotaenia bimaculata. The infestation rate for all species was 6.8 infested tires per 10,000 tires (wet and dry) inspected. Aedes albopictus larvae were most frequently collected, occurring at a rate of 20 infested wet tires per 10,000 inspected.