Assessing the cadmium content of cacao crops in Arauca, Colombia.

The district of Arauca is the second-largest producer of cacao in Colombia. However, despite its quality, it faces issues for export due to levels of cadmium (Cd) higher than the regulatory thresholds. A central question is how it may impact agricultural performance in the presence of Cd in cacao and chocolates. This study quantified Cd in cacao plantations from Arauca. Thus, 180 farms were assessed in the municipalities of Arauquita, Fortul, Saravena, and Tame. Five sample types (soil, irrigation channel sediment, soil litter, cacao seeds, and chocolates) were assessed for Cd. As a technological innovation, the new MXRF technology was used for Cd in chocolates. The sequence of Cd content was soil litter > chocolate > soils > cacao seeds > irrigation-channel sediment. A gradient north-south of Cd content in soil was observed, where highest content was found in farms near the Arauca River, and lower farther away. In irrigation channel sediment, Cd levels averaged 0.07 mg kg-1. The Cd content in cacao seeds was 0.78 mg kg-1 on average. Cd content in chocolates was above the threshold (1.10 mg kg-1 on average, including several cacao mass percentages). These artisanal chocolate bars produced by single farms were near the limit of Cd set by the European Union (up to 0.8 mg kg-1). Therefore, mixing beans from different farms could reduce their Cd content. The present study underscores the complexity of Cd distribution, emphasizing the importance of integrating soil, crop, and landscape features in managing and mitigating Cd levels in cacao.

FPGA-Based High-Performance Embedded Systems for Adaptive Edge Computing in Cyber-Physical Systems: The ARTICo³ Framework.

Cyber-Physical Systems are experiencing a paradigm shift in which processing has been relocated to the distributed sensing layer and is no longer performed in a centralized manner. This approach, usually referred to as Edge Computing, demands the use of hardware platforms that are able to manage the steadily increasing requirements in computing performance, while keeping energy efficiency and the adaptability imposed by the interaction with the physical world. In this context, SRAM-based FPGAs and their inherent run-time reconfigurability, when coupled with smart power management strategies, are a suitable solution. However, they usually fail in user accessibility and ease of development. In this paper, an integrated framework to develop FPGA-based high-performance embedded systems for Edge Computing in Cyber-Physical Systems is presented. This framework provides a hardware-based processing architecture, an automated toolchain, and a runtime to transparently generate and manage reconfigurable systems from high-level system descriptions without additional user intervention. Moreover, it provides users with support for dynamically adapting the available computing resources to switch the working point of the architecture in a solution space defined by computing performance, energy consumption and fault tolerance. Results show that it is indeed possible to explore this solution space at run time and prove that the proposed framework is a competitive alternative to software-based edge computing platforms, being able to provide not only faster solutions, but also higher energy efficiency for computing-intensive algorithms with significant levels of data-level parallelism.

A Deep Learning Approach for Fear Recognition on the Edge Based on Two-Dimensional Feature Maps.

Applying affective computing techniques to recognize fear and combining them with portable signal monitors makes it possible to create real-time detection systems that could act as bodyguards when users are in danger. With this aim, this paper presents a fear recognition method based on physiological signals obtained from wearable devices. The procedure involves creating two-dimensional feature maps from the raw signals, using data augmentation and feature selection algorithms, followed by deep learning-based classification models, taking inspiration from those used in image processing. This proposal has been validated with two different datasets, achieving, in WEMAC, WESAD 3-classes, and WESAD 2-classes, F1-score results of 78.13%, 88.07%, and 99.60%, respectively, and 79.90%, 89.12%, and 99.60% in accuracy. Furthermore, the paper demonstrates the feasibility of implementing the proposed method on the Coral Edge TPU device, prepared to make inferences on the edge.

A hurdle strategy based on the combination of non-thermal treatments to control diarrheagenic E. coli in cheese.

This study aimed to assess the efficacy of a multi-hurdle process combining mild High Hydrostatic Pressure (HHP) treatments and Thyme Oil (TO) edible films as a non-thermal method to combat pathogenic E. coli (aEPEC and STEC) in raw cow's-milk cheese stored at 7 °C and packaged under modified atmosphere. Changes in headspace atmosphere of cheese packs and treatment effects on Lactic Acid Bacteria (LAB) counts and diarrheagenic E. coli strains (aEPEC and STEC) were evaluated over a 28 d storage period. The results demonstrated that the combined treatment exhibited the most significant antimicrobial effect against both strains compared to individual treatments, achieving reductions of 4.30 and 4.80 log cfu/g after 28 d of storage for aEPEC and STEC, respectively. Notably, the synergistic effect of the combination treatment resulted in the complete inactivation of intact cells for STEC and nearly completed inactivation for aEPEC by the end of the storage period. These findings suggest that the combination of HHP with selected hurdles could effectively enhance microbial inactivation capacity, offering promising alternatives for improving cheese safety without affecting the starter microbiota.

Genetic characterization of atypical enteropathogenic Escherichia coli isolates from ewes' milk, sheep farm environments, and humans by multilocus sequence typing and pulsed-field gel electrophoresis.

A collection of 81 isolates of enteropathogenic Escherichia coli (EPEC) was obtained from samples of bulk tank sheep milk (62 isolates), ovine feces (4 isolates), sheep farm environment (water, 4 isolates; air, 1 isolate), and human stool samples (9 isolates). The strains were considered atypical EPEC organisms, carrying the eae gene without harboring the pEAF plasmid. Multilocus sequence typing (MLST) was carried out with seven housekeeping genes and 19 sequence types (ST) were detected, with none of them having been previously reported for atypical EPEC. The most frequent ST included 41 strains isolated from milk and human stool samples. Genetic typing by pulsed-field gel electrophoresis (PFGE) resulted in 57 patterns which grouped in 24 clusters. Comparison of strains isolated from the different samples showed phylogenetic relationships between milk and human isolates and also between milk and water isolates. The results obtained show a possible risk for humans due to the presence of atypical EPEC in ewes' milk and suggest a transmission route for this emerging pathogen through contaminated water.

Cacao (Theobromacacao L.) climate zones and its associated agrobiodiversity in Arauca, Colombia.

Background: Cacao (Theobromacacao L) is one of the most relevant crops in terms of economy and social rural development in Colombia. Cacao is also an important crop due to its potential to replace illicit crops and it is related to less deforestation and preserves the biodiversity. There are several cacao districts in Colombia, one of these being Arauca. The Department of Arauca is the second largest cocoa producing region in Colombia; however, it is heavily affected by armed conflict. To raise the knowledge and technology available in the region, integrating data on the occurrence of cacao farms with climatic variables becomes a powerful socioeconomic mapping tool for maintaining agrobiodiversity and food security in the region. Consequently, this type of agrodiversity data and agroclimatic approaches help to better manage agrobiodiversity, as in the cacao region of Arauca. These tools are even more relevant in biodiverse regions, such as flooded savannahs and tropical forest ecosystems, which are currently undergoing drastic changes due to agricultural expansion and climate change. One of the knowledge gaps in Colombia´s cacao regions is that there are currently no agroclimatic maps made with a social and scientific approach. This study aimed to provide a database of the spatial distribution of cacao farms in Arauca, as well as agroclimatic maps that identify and locate cacao climate regions in Arauca. We also present a presence-only matrix consisting of twenty-six tree species, or agrobiodiversity, distributed across the study region and specifically associated with the cacao forestry systems in Arauca. New information: We present the first database of both climate and agrobiodiversity data related to cacao farms in Arauca, developed with a research and socioeconomic vision that generated a novel approach for the agroclimatic zoning of cocoa in the Arauca Region and Colombia. Using 1,538 cacao farms at the regional scale, we identified two national and six regional-scale climate and soil regions. The selection at the local scale allowed us to classify 180 cacao farms comprising nine agroclimatic clusters in Arauca. We found twenty-six tree species distributed across the cacao climate zones. This dataset and its related maps also represent the agrobiodiversity of cultivated cacao locally. This is the most complete climate and agrobiodiversity dataset of cacao farms distribution in one of the top cocoa-producing regions in the country. These outputs are crucial because they constitute a baseline for developing research in the biodiversity of agroforestry systems, pests and diseases, pollutant presence, genetics, post-harvest processing and cocoa quality and safety.

Using SRAM based FPGAs for power-aware high performance wireless sensor networks.

While for years traditional wireless sensor nodes have been based on ultra-low power microcontrollers with sufficient but limited computing power, the complexity and number of tasks of today's applications are constantly increasing. Increasing the node duty cycle is not feasible in all cases, so in many cases more computing power is required. This extra computing power may be achieved by either more powerful microcontrollers, though more power consumption or, in general, any solution capable of accelerating task execution. At this point, the use of hardware based, and in particular FPGA solutions, might appear as a candidate technology, since though power use is higher compared with lower power devices, execution time is reduced, so energy could be reduced overall. In order to demonstrate this, an innovative WSN node architecture is proposed. This architecture is based on a high performance high capacity state-of-the-art FPGA, which combines the advantages of the intrinsic acceleration provided by the parallelism of hardware devices, the use of partial reconfiguration capabilities, as well as a careful power-aware management system, to show that energy savings for certain higher-end applications can be achieved. Finally, comprehensive tests have been done to validate the platform in terms of performance and power consumption, to proof that better energy efficiency compared to processor based solutions can be achieved, for instance, when encryption is imposed by the application requirements.

Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae in Fresh Produce.

Fresh vegetables are an essential part of a healthy diet, but microbial contamination of fruits and vegetables is a serious concern to human health, not only for the presence of foodborne pathogens but because they can be a vehicle for the transmission of antibiotic-resistant bacteria. This work aimed to investigate the importance of fresh produce in the transmission of extended-spectrum ß-lactamases (ESBL)-producing Enterobacteriaceae. A total of 174 samples of vegetables (117) and farm environment (57) were analysed to determine enterobacterial contamination and presence of ESBL-producing Enterobacteriaceae. Enterobacterial counts above the detection limit were found in 82.9% vegetable samples and 36.8% environmental samples. The average count was 4.2 log cfu/g or mL, with a maximum value of 6.2 log cfu/g in a parsley sample. Leafy vegetables showed statistically significant higher mean counts than other vegetables. A total of 15 ESBL-producing isolates were obtained from vegetables (14) and water (1) samples and were identified as Serratia fonticola (11) and Rahnella aquatilis (4). Five isolates of S. fonticola were considered multi-drug resistant. Even though their implication in human infections is rare, they can become an environmental reservoir of antibiotic-resistance genes that can be further disseminated along the food chain.

Behaviour of Non-O157 STEC and Atypical EPEC during the Manufacturing and Ripening of Raw Milk Cheese.

This study was carried out to assess the survival of Shiga toxin-producing E. coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) during the traditional manufacturing and ripening of Spanish hard cheese from raw cow's milk. Milk samples were spiked with up to 3.1-3.5 log cfu/mL of one strain of STEC (O140:H32 serotype) and one of aEPEC (serotype O25:H2). The first steps of cheesemaking allow for a STEC and aEPEC increase of more than 1 log cfu/mL (up to 4.74 log cfu/g and 4.55 log cfu/g, respectively). After cheese pressing, a steady reduction of both populations was observed, with the STEC strain being more sensitive. The studied pathogenic E. coli populations decreased by 1.32 log cfu/g in STEC and 0.59 log cfu/g in aEPEC in cheese ripened during a minimum period of 60 d. Therefore, a moderate contamination by these diarrhoeagenic E. coli pathotypes, in particular, with aEPEC, on cheese manufactured from raw milk may not be totally controlled through the cheesemaking process and during a maturation of 90 d. These findings remark the importance of improvement in bacteriological quality of raw milk and cross-contamination prevention with diarrhoeagenic E. coli in the dairy industry.

Effect of different storage conditions on E. coli O157:H7 and the indigenous bacterial microflora on lamb meat.

Lamb chops inoculated with 2.23-2.83 log cfu/g of E. coli O157:H7 strain NCTC 12900 were packed in air (AP), vacuum (VP), and two modified atmospheres (MAP) consisting of 100% CO2 and a commercial mixture of 35% CO2/35% O2/30% N2. All samples (initial total counts <3.5 log cfu/g) were stored in a commercial cold storage facility set at 4 degrees C and one AP trial also at 12+/-1 degrees C in a temperature controlled incubator. Pathogen and indigenous flora evolution, physicochemical and sensory changes, surface packages temperature and MAP gas composition were monitored throughout the lamb meat shelf life. Temperature monitoring revealed that during chilled storage packed chops exceeded 7 degrees C about 3% of the time for periods of 10-20 min at 6 h intervals corresponding to defrosting cycles. In AP samples under these conditions, the E. coli O157:H7 strain had an overall increase of 0.48 log cfu/g by day 12. This increase, which may be regarded as an artefact of the sampling procedure, might also be a response to fluctuating temperatures. Regardless of rapid proliferation of the background microflora on AP lamb meat kept at 12+/-1 degrees C, the pathogen significantly increased by 2.35 log cfu/g after nine days. There was a slight decrease (0.20 log cfu/g) of the pathogen numbers after four weeks cold storage in VP despite a significant increase in lactic acid bacteria (LAB). With a relatively small outgrowth of LAB, chilled storage in 100% and 35% CO2 resulted in significant differences compared to similar conditions in air (decrease from initial numbers of 0.80 and 0.45 log cfu/g, respectively). Our data confirm the importance of effective temperature control to prevent pathogen growth on raw meat and also that contaminated meat remains hazardous regardless of refrigeration and protective packaging. Further studies are needed to determine the behaviour of E. coli O157:H7 at temperatures that fluctuate around the minimum for growth.

Cell-associated hemolytic activity in environmental strains of Plesiomonas shigelloides expressing cell-free, iron-influenced extracellular hemolysin.

Hemolysis is a means of providing pathogenic bacteria with heme iron in vivo. In a previous work, iron-influenced hemolytic activity against sheep erythrocytes was detected in cell-free supernatants, but not in the cell fraction of two environmental Plesiomonas shigelloides strains incubated without shaking. Both strains have the hugA gene, which encodes an outer membrane receptor required for heme iron utilization. The present study was undertaken to investigate the expression of a second hemolytic activity detected during aerated incubation in normal and iron-depleted tryptone soya broth (id-TSB). An agar overlay procedure and doubling dilution titrations were employed to detect the hemolytic activity against several erythrocyte species. The kinetics of growth and hemolytic activity were assayed at 35 degrees C in aerated normal and id-TSB and salmon extract. Overlaid colonies showed a cell-associated beta-hemolytic activity within 4 h. For aerated cell-free supernatants, titers above 16 were not attained until 30 to 48 h of incubation; the best activity was noted with dog and mouse erythrocytes. After 24 h of aerated incubation, sonicated cells yielded high hemolytic activity against dog erythrocytes without activity in supernatants, but after 48 h, only 28 to 30% of the total activity remained cell associated. The hemolytic factor was released in broths during the death phase. Hemolytic activity was not detected in fish extract. This and other studies suggest that P. shigelloides may produce at least two hemolytic factors, their expression and detection being influenced by environmental growth conditions and testing procedures. The overlay assay appears to be the best routine method for detecting hemolytic activity in P. shigelloides.

Molecular and phenotypic typing of Staphylococcus aureus isolates from rabbit meat.

Twenty-six Staphylococcus aureus isolates were recovered from rabbit carcasses and cuts during a period of seven months. Samples from 51 different animals, flocks and farms were obtained from five establishments in four Spanish provinces. To determine their diversity and possible origin, isolates were typed by three molecular and three phenotypic methods. PFGE, with the highest discrimination index (D=0.966), identified 19 patterns (more than one band difference) and 10 types (more than three band differences). Based on > or = 90% similarity, RAPD (D=0.877) produced nine patterns while ribotyping (D=0.786) produced seven types. On the basis of biotyping (D=0.644), 11 isolates belonged to human ecovars and 15 to the non-host-specific crystal violet type C (NHS CV:C) biotypes. By direct phage typing (D=0.761), 17 isolates were lysed by human phages into groups II (8 isolates), III (5 isolates), I/III (2 isolates) and V (2 isolates). The overall resistance to antimicrobials (D=0.783) was 76.9%, with most isolates being resistant to tetracycline (61.5%) and penicillin G (26.9%). PFGE showed that samples from each processing plant carried different S. aureus types, some of them persisting over time. There also was evidence of interestablishment dissemination of genetically related clones, most of them belonging to the PFGE type A and phenotype "NHS CV:C biotypes-3A/3C/55/71 phage type", which is highly virulent for European commercial rabbitries. The ubiquity of the virulent phenotype, as well as the high incidence of resistance to antibiotics with application in human medicine, is a matter of concern in public and animal health.

Occurrence of Plesiomonas shigelloides in displayed portions of saltwater fish determined by a PCR assay based on the hugA gene.

PCR primers were designed and used to amplify a 435-bp fragment from the Plesiomonas shigelloides hugA gene. The PCR assay combined with a non-selective enrichment step proved to be a reliable procedure for P. shigelloides detection in fish meat. The incidence of this bacterium was investigated in 52 lots of pre-packed saltwater fish portions (conger, swordfish, sole, grouper, whiting and halibut) displayed at two hypermarkets by a conventional two-step procedure and the PCR assay. Using the former, P. shigelloides was isolated from three lots of grouper fillets and one lot of halibut fillets. When PCR was performed with non-selective enriched cultures of fish portions, amplification products were obtained from samples that were positive by the culturing method and from eight additional lots of grouper fillets that gave negative results with the conventional procedure. After a secondary enrichment in tetrathionate broth without iodine, all PCR-positive non-selective enrichments yielded P. shigelloides colonies. Overall, P. shigelloides was found in 23% of the examined lots of marine fish (11 of grouper and one of halibut).

Rabbit meat as a source of bacterial foodborne pathogens.

Even though worldwide production of rabbit meat is >1,000,000 tons, little information is available for rabbit meat microbiology. This study provides data on the prevalence of Salmonella, Escherichia coli O157:H7, Yersinia enterocolitica, Listeria spp., motile Aeromonas spp., and Staphylococcus aureus on rabbit meat. A total of 24 rabbit carcasses from two abattoirs and 27 rabbit meat packages from supermarket displays were examined. In addition to culturing methods, associated virulence genes were investigated by PCR in suspect isolates and samples. Neither Salmonella nor E. coli O157:H7 was detected. All samples were negative for virulence-associated invA, stx1, and stx2 genes. At one abattoir, two carcasses (3.9%) carried Y. enterocolitica yst-, and two were positive for the yst gene, although viable Y. enterocolitica cells were not recovered from these samples. Seven samples (13.7%) were contaminated with Listeria. Of them, three were positive for hly and iap genes (Listeria monocytogenes hly+ / iap+), two carried Listeria seeligeri, one carried Listeria ivanovii, and one carried Listeria innocua. For detectable motile Aeromonas spp. (average count, 1.77 +/- 0.62 log CFU/g), the contamination rate was 35.3%, although ca. 90% of the samples were positive for the aerA and/or hlyA genes. The majority of aeromonad isolates were Aeromonas hydrophila aerA+ / hlyA+. Aeromonas caviae, Aeromonas popoffii, Aeromonas schubertii, and the two biovars of Aeromonas veronii were also isolated. The prevalence of S. aureus contamination (average count, 1.37 +/- 0.79 log CFU/g) was 52.9%. Among 27 S. aureus isolates, two harbored genes for staphylococcal enterotoxin B (seb), and two harbored genes for staphylococcal enterotoxin C (sec). The remaining isolates were negative for sea, seb, sec, sed, and see.

Genetic characterization of Shiga toxin-producing Escherichia coli (STEC) and atypical enteropathogenic Escherichia coli (EPEC) isolates from goat's milk and goat farm environment.

The aim of this study was to characterize a collection of 44 Shiga toxin-producing (STEC) and enteropathogenic Escherichia coli (EPEC) isolated from goat milk and goat farm environment. Of the 19 STEC isolates, five (26.3%) carried the stx1 gene, four (21.1%) the stx2 gene and 10 (52.6%) presented both stx genes. Six (31.6%) STEC strains were eae-positive and belonged to serotypes related to severe human disease (O157:H7 and O5:HNM). Another seven STEC strains were of serotype O146:H21 and three of serotype O166:H28, also linked to human disease. The STEC strains isolated from goat milk were of serotypes potentially pathogenic for humans. All the 25 EPEC isolates were considered atypical (aEPEC) and one aEPEC strain was of serotype O26:H11, a serotype frequently isolated in children with diarrhea. Multilocus sequence typing (MLST) was carried out with seven housekeeping genes and 23 sequence types (ST) were detected, 14 of them newly described. Twelve STs grouped STEC isolates and 11 STs grouped EPEC isolates. Genetic typing by pulsed field gel electrophoresis (PFGE) resulted in 38 patterns which grouped in 10 clusters. Well-defined groups were also observed for strains of pathogenic serotypes. In conclusion, strains of STEC and aEPEC belonging to serotypes related to severe human disease have been detected in goat milk and the goat farm environment. Ruminants are an important reservoir of STEC strains and the role of these animals as carriers of other pathogenic types of E. coli seems to be an emerging concern.

Incidence, radioresistance, and behavior of Psychrobacter spp. in rabbit meat.

The relative incidence of Psychrobacter spp. in rabbit meat, the radioresistance of these bacteria, and the growth of nonirradiated and irradiated psychrobacter isolates, alone and in coculture, during chilled storage of inoculated sterile rabbit meat was investigated. Psychrobacter spp. accounted for 4.2% of the storage psychrotrophic flora of 30 rabbit carcasses. The radiation D10-values of 10 Psychrobacter isolates, irradiated at 4 degrees C in minced rabbit meat, ranged from 0.8 to 2.0 kGy, with significant (P < 0.05) differences among strains. Over 12 days of storage at 4 degrees C, pure cultures of two nonirradiated psychrobacter strains (D10 = 2 kGy) were capable of substantial increases (up to 3 log CFU/g) in sterile rabbit meat, but when the fastest growing strain was cocultured with Pseudomonas fluorescens and Brochothrix thermosphacta isolates, maximum cell densities and growth rates were significantly (P < 0.01) lower. After irradiation (2.5 kGy) of pure cultures in sterile rabbit meat, surviving cells of both Psychrobacter strains decreased for a period of 5 to 7 days and then resumed multiplication that, at day 12, resulted in a similar increase (1.6 to 1.7 log CFU/g) over initial survivor numbers. When irradiated in combination with the spoilage bacteria, one of the strains required 12 days to reach initial numbers. In conclusion, Psychrobacter spp. are radioresistant nonsporeforming bacteria with a low relative incidence among the storage flora of rabbit meat, unable to compete with food spoilage bacteria in this ecosystem and apparently not a major contributor to the spoilage of rabbit meat after irradiation.

Development of the aerobic spoilage flora of chilled rabbit meat.

Even though worldwide production of rabbit meat is over 1,000,000ton, little information is available on rabbit meat microbiology. This paper reports on the microflora developing on chill-stored rabbit carcasses. Four different lots of 24h post-mortem rabbit carcasses dressed and kept at 0°C in a medium-size abattoir were collected and evaluated for sensory, physicochemical and microbiological changes during aerobic storage at 3±1°C. Mean initial pH value (pH(24)), extract-release volume (ERV) and lactate content of Biceps femoris muscle, were 6.26±0.20, 13.50±3.50ml and 0.70±0.07%, respectively. As with other muscle foods kept chilled in air, pH increased and ERV and lactate decreased as storage progressed. Initial levels (logcfu/g) of aerobes (APC), psychrotrophic flora, Pseudomonas spp., Brochothrix thermosphacta, lactic acid bacteria, Enterobacteriaceae and yeasts were 4.76±0.31, 4.81±0.81, 3.39±1.12, 2.01±0.92, 2.76±0.51, 0.49±0.45 and 3.46±0.32, respectively. Pseudomonads, most of them fluorescent, and to a lesser extent B. thermosphacta and yeasts grew faster than the remaining microorganisms and became predominant at the end of the shelf life. Carcasses spoiled when mean APC, psychrotrophic and pseudomonads numbers were ca. 8logcfu/g, their mean shelf life being estimated at 6.8 days. A lot of DFD-like rabbit carcasses, with higher pH and lower ERV values but similar microbial loads to normal meat, developed a strong putrid odour after 4 days.

Microbiological Examination of Bulk Tank Goat's Milk in the Castilla y León Region in Northern Spain.

The purpose of the study was to evaluate the microbiological status (mesophilic aerobic microorganism counts) of 68 samples of bulk tank goat's milk and determine the risk associated with the foodborne pathogens Staphylococcus aureus, enteropathogenic and Shiga toxin-producing Escherichia coli, and Cronobacter sakazakii. Most samples (83.8%) complied with the limits of mesophilic aerobe counts set in the European Union for milk of species other than cows. A total of 144 isolates of coagulase-positive staphylococci were characterized, and 11 (7.6%) of them carried staphylococcal enterotoxin (SE) genes of the classical types (encoding SEA to SEE), distributed as follows: 4 carried the SEA gene, 1 the SEB gene, and 6 the SED gene. C. sakazakii was not detected in any sample. Regarding detection of E. coli virulence-related genes in enriched milk samples, 12 milk samples were positive only for the presence of stx genes, 4 were positive for both stx and eae genes, and 20 were negative for stx amplification and positive for eae amplification. Seven enteropathogenic E. coli and 9 Shiga toxin-producing E. coli isolates (one of them of serogroup O157) were recovered. In conclusion, goat's milk produced on farms in Castilla y León is generally in accordance with European Union standards, but the presence of pathogenic E. coli isolates indicates that the consumption of raw goat's milk may pose a risk to public health.

Numbers and types of microorganisms in vacuum-packed cold-smoked freshwater fish at the retail level.

Fifty-four packages (each one belonging to a different lot) of vacuum-packed cold-smoked salmon (30) and trout (24) produced by six Spanish smokehouses were obtained at retail level after 3 weeks storage at 2+/-1 degrees C. Sensorial, chemical, physicochemical and microbiological characteristics were examined. Overall, pH, a(w), salt content in water phase, aerobic plate counts at 30 and 25 degrees C. levels of Enterobacteriaceae, lactic acid bacteria (LAB), fungi and presumptive aeromonads and staphylococci are in agreement with available data on lightly preserved fish products. Psychrotrophic clostridia ranged between 1.71 and 2.21 log CFU/g. Levels of ethanol were highly variable and not significantly related (p > 0.05) to sensory scores or to microbial numbers. Salmonella, Escherichia coli and Listeria monocytogenes were not detected in any sample. Listeriae other than L. monocytogenes were isolated from three packages. Levels of Staphylococcus aureus lower than 4 log CFU/g were also found in three packages. Among 377 bacteria randomly isolated from aerobic 25 degrees C plate counts, LAB predominated, with Carnobacterium (C. piscicola) and Lactobacillus (eight species) being the genera most frequently found. The second and third major groups were Enterobacteriaceae and Micrococcaceae, respectively. Proteus vulgaris, P. mirabilis and Serratia liquefaciens were dominant among Enterobacteriaceae and coagulase-negative staphylococci among Micrococcaceae. Minor microbial groups such as aerobic gram-negative bacilli (Acinetobacter; Moraxella and Pseudomonas), Brochothrix, Aeromonas, Bacillus and Vibrio constituted less than 17% of the total flora.

Hemolytic and proteolytic activities of Aeromonas hydrophila and Aeromonas veronii biovar sobria in broth and salmon extract at different temperatures.

Expression of hemolytic and proteolytic activities throughout the growth cycle was investigated with two enterotoxic aeromonad strains assigned to the species Aeromonas hydrophila and Aeromonas veronii biovar sobria. Although growth kinetic data were dependent on strain, temperature, and substrate, maximum populations attained were higher than 9 log CFU/ml in aerated tryptone soya broth plus yeast extract (TSBYE) and salmon extract within the range 4 to 28 degrees C. For both strains in TSBYE, variable amounts of hemolytic activity were first detected at any temperature when aeromonad counts were over 9 log CFU/ml. Afterwards, this activity increased up to similar levels (109 to 112 hemolytic units per ml) without a significant increase in populations. Salmon extract supported hemolysin synthesis at 28 but not 4 degrees C. Proteolytic activity of the A. hydrophila strain was only expressed in salmon extract at 28 degrees C, whereas A. veronii biovar sobria did at 28 degrees C in both substrates and at 10 degrees C in TSBYE.