Presence Of Enterobacteriaceae Research Articles

Prevalence of Enterobacteriaceae in Camel, Cattle, and Sheep Carcasses at Slaughterhouses and Butcher Shops

Enterobacteriaceae can contaminate meat during various processing stages, including slaughter, evisceration, and b utchering, potentially causing foodborne illnesses. The purpose of this study was to investigate the prevalence of Enterobacteriaceae in carcass samples obtained from slaughterhouses and meat cuts collected from butcher shops. A total of 120 samples of camel, cattle, and sheep meat were analyzed for microbial contamination and isolates were identified using the PCR test. Total viable count ranged from 4.91 to 5.37 Log10 CFU/g in slaughterhouses and butcher shops. E. coli dominated, with 84 out of the 120 samples (70%) contaminated, where contamination was highest in camel meat and lowest in sheep meat with 100% and 30% of contaminated samples, respectively. Salmonella was confirmed in 40% of camel, 47.5% of cattle, and 32.5% of sheep samples. In addition, twenty-five Enterobacteriaceae strains belonging to 19 different genera were detected in the meat samples. The highest occurrence was in the sheep samples with 15 different genera followed by the camels and the cattle samples with 14 different genera each. The presence of Enterobacteriaceae in camel, cattle, and sheep carcasses raises significant concerns regarding food safety. Adherence to good hygiene practices throughout animal slaughtering is crucial to minimize the risk of infection and transmission and ensure food safety.

Salmonella spp., Escherichia coli and Enterobacteriaceae Control at a Pig Abattoir: Are We Missing Lairage Time Effect, Pig Skin, and Internal Carcass Surface Contamination?

To provide meat safety and consumer protection, appropriate hygiene control measures at an abattoir are required. This study aimed to evaluate the influence of visual fecal contamination level (VFCL) and lairage time (LT) on pig skin (PS) and external (ECS) and internal (ICS) carcass surfaces. The presence of Enterobacteriaceae, Escherichia coli (E. coli) and Salmonella in PS, ECS, and ICS were evaluated. A total of 300 paired samples were collected from 100 pigs. Results underlined the importance of the skin (Enterobacteriaceae: 3.27 ± 0.68 log CFU/cm2; E. coli: 3.15 ± 0.63 log CFU/cm2; Salmonella: 21% of samples) as a direct or indirect source of carcass contamination. Although VFCL revealed no significant effect (p > 0.05), the increase of LT had a significant impact (p < 0.001) on Enterobacteriaceae and E. coli levels across all analysed surfaces, and Salmonella presence on ICS (p < 0.01), demanding attention to LT. Also, the ICS showed a higher level of these bacteria compared to ECS. These results highlight the need of food business operators to consider ICS as an alternative area to sample for Salmonella, as a criterion for process hygiene based on EC Regulation No. 2073/2005, and as a potential contamination source to be integrated in the hazard analysis critical control point (HACCP) plans.

Sous-vide processing of silver carp: Effect of processing temperature and cold storage duration on the microbial quality of the product as well as modeling by artificial neural networks.

Silver carp (Hypophthalmichthys molitrixi) was processed by sous-vide method at different temperatures (60, 65, 70, and 75°C). Then, the microbiological quality of the processed samples was monitored during cold storage (4°C) for 21 days. The target microorganisms were Enterobacteriaceae, Lactic Acid bacteria (LAB), Pseudomonas, Psychrotrophs, and total viable count (TVC). In samples processed at 75°C, the presence of Enterobacteriaceae, Pseudomonas and Psychrotrophs were not detectable up to 15 days of storage and lactic acid bacteria were not detectable even at the end of the storage period. A radial basis function neural network (RBFNN) model was established to predict the changes in the microbial content of silver carp. In this step, the relationship between processing temperature and storage duration on microbial growth was modeled by ANNs (artificial neural networks). The optimal ANN topology for modeling Enterobacteriaceae, Pseudomonas, and Psychrotroph contained 9 neurons in the hidden layer, but it contained 15 and 14 neurons for TVC and LAB, respectively. By experimenting with the temperature of -80°C, it was revealed that the obtained ANN model has a high potential for prediction.

Are Enterobacteriaceae and Enterococcus Isolated from Powdered Infant Formula a Hazard for Infants? A Genomic Analysis.

Powdered infant formulas (PIF) are the most used dietary substitutes that are used in order to supplement breastfeeding. However, PIF are not sterile and can be contaminated with different microorganisms. The objective of this study was to genomically characterize Enterobacteriaceae (ENT) and Enterococcus strains that were isolated from PIF. Strains were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and whole-genome sequencing (WGS). Genomic typing, detection of virulence, and resistance profiles and genes were performed with the Ridom SeqSphere+ software; the comprehensive antibiotic resistance database (CARD) platform; ResFinder and PlasmidFinder tools; and by the disk diffusion method. Nineteen isolates from PIF were analyzed, including ENT such as Kosakonia cowanii, Enterobacter hormaechei, Franconibacter helveticus, Mixta calida, and lactic acid bacteria such as Enterococcus faecium. The strains exhibited resistance to beta-lactams, cephalosporins, and macrolides. Resistance genes such as AcrAB-TolC, marA, msbA, knpEF, oqxAB, fosA, blaACT-7, blaACT-14,qacJ, oqxAB,aac(6')-Ii, and msr(C); and virulence genes such as astA, cheB, cheR, ompA ompX, terC, ironA, acm, and efaAfm, adem were also detected. All the analyzed strains possessed genes that produced heat-shock proteins, such as IbpA and ClpL. In PIF, the presence of ENT and Enterococcus that are multiresistant to antibiotics-together with resistance and virulence genes-pose a health risk for infants consuming these food products.

Enterobacteriaceae dan Personel Hygiene Ibu Terhadap Kejadian Diare Anak 5 Tahun di Kecamatan Ratu Samban Kota Bengkulu

Diarrheal disease is one of the environment-based diseases and is closely related to personal hygiene conditions and the presence of Enterobacteriaceae in food. Diarrheal diseases can also spread from person to person, exacerbated by poor personal hygiene. Food is another major cause of diarrhea when prepared or stored in unhygienic conditions. Unsafe storage and handling of household water is also an important risk factor. Personal habits that do not want to wash hands with soap before carrying out their activities, as well as other behaviors that do not reflect a healthy lifestyle can cause diarrhea. This study aims to determine the relationship between enterobacteriaceae in food and maternal personal hygiene with the incidence of diarrhea in children less than 5 years old in Ratu Samban District, Bengkulu City. This research is an analytic survey research with a cross-sectional approach. From the research data, statistical test results were obtained with chi square, there was no significant relationship between Enterobacteraiceae in food and maternal hygiene personnel with the incidence of diarrhea in children less than 5 years old in Ratu Samban District, Bengkulu City in 2022 where the p value &gt; from 0.05.

Risk Factors of Infections Due to Multidrug-Resistant Gram-Negative Bacteria in a Community Hospital in Rural Thailand.

Antimicrobial resistance is a major public health concern globally. The most serious antimicrobial resistance problem among pathogenic bacteria is multidrug resistance (MDR). The objectives of this study were to investigate the risk factors of MDR infections and to develop a risk assessment tool for MDR Gram-negative bacteria (MDR-GNB) infections at a community hospital in rural Thailand. The study revealed 30.77% MDR-GNB among GNB strains. The most common MDR-GNB strains were 63.02% for Escherichia coli and 11.46% for Klebsiella pneumoniae. A case-control study was applied to collect clinical data between January 2016 and December 2020. Univariate logistic regression and multivariate logistic regression were used to analyze the risk factors for MDR-GNB and a risk assessment score for each factor was determined based on its regression coefficient. The risk factors for MDR-GNB infections were as follows: the presence of Enterobacteriaceae that produce extended-spectrum beta-lactamase (ESBL) (ORAdj. 23.53, 95% CI 7.00-79.09), infections occurring within the urinary tract (ORAdj. 2.25, 95% CI 1.44-3.53), and patients with a history of steroid usage (ORAdj. 1.91, 95% CI 1.15-3.19). Based on the assigned risk scores for each associated factor, the newly developed risk assessment tool for MDR-GNB infections achieved 64.54% prediction accuracy (AUC-ROC 0.65, 95% CI 0.61-0.68), demonstrating that the tool could be used to assess bacterial infection cases in community hospitals. Its use should provide practical guidance on MDR evaluation and prevention. This study was part of an antibiotic stewardship program; the study surveyed antibiotic-resistant situations in a hospital and implemented an effective risk assessment tool using key risk factors of MDR-GNB infections.

Determination of In-door Air Quality by Estimating Microbiological Load from the Ambient Air among Basic Health Units in the City of Januaria, Brazil - A Cross Sectional Study

Aims: Poor air quality can compromise the health and recovery of patients and even compromise the quality of life and productivity of health professionals, affecting the speed of recovery of patients and allowing the occurrence of nosocomial infections. The present work evaluated the microbiological quality of ambient air in Basic Health Units (BHU), determining the degree of microbiological safety for the population served.&#x0D; Study Design: This study was conducted with triplicate evaluation of samples of ambient air.&#x0D; Place and Duration of Study: This study was conducted in the city of Januária, state of Minas Gerais, Brazil, between March 2016 and July 2016.&#x0D; Methodology: Environmental air samples were collected in five BHU in the urban area of the city, evaluating the contamination by aerophilic mesophilic microorganisms, enterobacteria, molds and yeasts, using the simple sedimentation technique in a Petri dish.&#x0D; Results: It was verified the presence of mesophilic aerobic bacteria and molds and yeasts in amounts higher than the recommendation used as a parameter for this study, indicating that the hygienic-sanitary conditions in the evaluated BHU are not adequate.&#x0D; Conclusion: The presence of Enterobacteriaceae in some evaluated sites suggests the possibility of the presence of pathogenic microorganisms, which may pose risks to the health of the population.

Assessment of Pseudomonas aeruginosa biofilm-forming capacities from drinking water in water vending machine

The establishment of P. aeruginosa with biofilm formation in water vending machines may cause serious health issues to the consumers and its emergence has led to the public’s concern. This study aimed to assess the quality of water vending machines and to evaluate the biological contaminant P. aeruginosa in biofilm capacities. The qualities of the drinking water from a total of fifteen (n = 15) water vending machines at Kota Samarahan were evaluated based on physical and chemical parameters including pH value, turbidity, total of carbon (TOC), total dissolved solid (TDS) and total suspended solid (TSS). The colonies Enterobacteriaceae has been morphology characterized through biochemical tests and P. aeruginosa bacteria was identify through the PCR method. The results of the physical and chemical parameters complies with the authority standard including turbidity values found in conformance with values were lower than 0.1 NTU. Morphological analysis with a total of 66.7% (n = 10) was detected with the presence of Enterobacteriaceae, and a total of 40% (n = 6) of the isolates were found to be P. aeruginosa. This study extended by assessing the potential strength of biofilm formation. The microtiter assay performed in a 96-well polystyrene microtiter plate showed that 83.33% (n = 5) of the bacterial isolates have moderate potential as biofilm producers, while only 16.67% (n = 1) isolates were non-adherent and showed no potential in producing biofilm. The highest OD isolates found occupying moderate biofilm strength was (mean = 0.217) and the lowest moderate biofilm strength was (mean = 0.136). In conclusion, the significance and impact of the study displayed the qualities of water vending machines complies with Food Act 1983, Regulation 360C and Malaysian Drinking Water Quality, Ministry of Health 1983. However, the presence of biological contaminants may raise consumer concerns. This study had successfully assessed the potential strength of P. aeruginosa biofilm collected from water vending machines. Further microbiological assessments should be perform continuously to predict and eliminate any future risks related to water vending machines.

Antimicrobial Resistant Bacteria in Health Care Facilities: Exploring Links With Water, Sanitation, and Hygiene in Gaza, Palestine

Background Antimicrobial resistance (AMR) is a growing global phenomenon; however, its link to water, sanitation, and hygiene (WASH) remains underexplored, particularly in health care facilities where humanitarian crises prevail. Objective This study aimed to identify AMR bacteria in samples collected from WASH services in 2 hospitals in Gaza and to investigate the presence of AMR genes. Methods A hospital-based cross-sectional study to detect and identify antimicrobial resistance bacteria was conducted. Random samples from water, wastewater, soap, and surface swabs (n=345) were collected from Al-Shifa and European Gaza hospitals and screened for the presence of Enterobacteriaceae, Pseudomonas, Enterococcus, and Staphylococcusaureus. Antimicrobial susceptibility, extended spectrum beta-lactamase (ESBL) production, carbapenem resistance, and AMR genes were investigated. Results High levels of bacterial contamination were detected in water and surface swab samples with an overall percentage of 34.1%. Moreover, 22% of the identified Enterobacteriaceae was positive for ESBL, and 14% was positive for modified Hodge test. Over 2/3 of isolated Enterobacteriaceae in water and wastewater samples was found to be resistant to amikacin, ceftazidime, ceftriaxone, and imipenem. All Enterobacteriaceae isolates from swab samples were found to be resistant to piperacillin-tazobactam, amikacin, ceftazidime, and ceftriaxone; 13.8% of S. aureus in water samples was methicillin resistant. The prevalence of ESBL genes among Enterobacteriaceae isolates was 25% OXA, 19.4% SHV, 2.8% KPC, 66.7% TEM, 41.7% blaCTXM, and 5.6% blaCTXM-3. For carbapenem-resistant gene (MDM), the prevalence among Enterobacteriaceae was 11.1%, and among Pseudomonas was 12.5%. The antibiotic susceptibility profile was also presented for Pseudomonas, Enterococcus, and S. aureus. Conclusions The results underline the level of contamination with AMR bacteria in WASH samples and highlight the need to consider the safety of WASH service at health care facilities as an essential aspect in the fight against the spread of AMR and to interrupt nosocomial transmission.

Age and Injury Size Influence the Magnitude of Fecal Dysbiosis in Adult Burn Patients.

Clinical studies have demonstrated that age 50 years or older is an independent risk factor associated with poor prognosis after burn injury, the second leading cause of traumatic injuries in the aged population. While mechanisms driving age-dependent postburn mortality are perplexing, changes in the intestinal microbiome, may contribute to the heightened, dysregulated systemic response seen in aging burn patients. The fecal microbiome from 22 patients admitted to a verified burn center from July 2018 to February 2019 was stratified based on the age of 50 years and total burn surface area (TBSA) size of ≥10%. Significant differences (P = .014) in overall microbiota community composition (ie, beta diversity) were measured across the four patient groups: young <10% TBSA, young ≥10% TBSA, older <10% TBSA, and older ≥10% TBSA. Differences in beta diversity were driven by %TBSA (P = .013) and trended with age (P = .087). Alpha diversity components, richness, evenness, and Shannon diversity were measured. We observed significant differences in bacterial species evenness (P = .0023) and Shannon diversity (P = .0033) between the groups. There were significant correlations between individual bacterial species and levels of short-chain fatty acids. Specifically, levels of fecal butyrate correlated with the presence of Enterobacteriaceae, an opportunistic gut pathogen, when elevated in burn patients lead to worsen outcomes. Overall, our findings reveal that age-specific changes in the fecal microbiome following burn injuries may contribute to immune system dysregulation in patients with varying TBSA burns and potentially lead to worsened clinical outcomes with heightened morbidity and mortality.

Serbian Traditional Goat Cheese: Physico-Chemical, Sensory, Hygienic and Safety Characteristics.

This research project aimed to investigate the physico-chemical, sensory, hygienic and safety characteristics of raw goat milk, whey, brine and traditional goat cheese during the ripening period of 28 days. Physico-chemical parameters included the determination of dry matter, fat, ash, protein, pH, water activity and NaCl content. The presence of Enterobacteriaceae and fungi was estimated on milk and cheese samples, and a sensory panel evaluated the products’ features and acceptability during ripening. The results show that the cheese under study belongs to the acid full-fat cheese group. A consumer panel attributed high scores to the goat cheese, until the 21st day of ripening. After this period, the overall features altered significantly, including augmented bitterness, odor intensification and the development of molds on the surface. The presence of fungi, associated with Enterobacteriaceae, suggests that the hygiene of the production processes needs to be improved. Regarding microbial safety, the detection of putative pathogens and antibiotic resistances recommend an active surveillance of traditional foods to avoid foodborne infections and/or the dissemination of resistant microorganisms along the food chain.

Short communication: Coliform Petrifilm as an alternative method for detecting total gram-negative bacteria in fluid milk

Postpasteurization contamination (PPC) of fluid milk remains a challenge for some dairy processors. Pseudomonas is the most common contaminant of fluid milk after pasteurization, and therefore methods to detect PPC should be inclusive of Pseudomonas and other gram-negative contaminants (e.g., coliform bacteria). Our objective was to compare the ability of 3M (St. Paul, MN) coliform and Enterobacteriaceae (EB) Petrifilm to detect total gram-negative bacteria with that of the standard method, crystal violet tetrazolium agar. To that end, we evaluated coliform Petrifilm, EB Petrifilm, and crystal violet tetrazolium agar to detect gram-negative bacteria in naturally contaminated samples of fluid milk. A total of 92 observations derived from shelf-life testing of 33 milk samples from 5 different processing facilities were evaluated for (1) presence of coliforms on coliform Petrifilm at both 24 and 48 h of incubation; (2) presence of any growth, regardless of gas production, on coliform Petrifilm at both 24 and 48 h of incubation; (3) presence of EB on EB Petrifilm at both 24 and 48 h of incubation; (4) presence of any growth, regardless of gas or acid production, on EB Petrifilm at both 24 and 48 h of incubation; and (5) presence of gram-negative bacteria on crystal violet tetrazolium agar after 48 h of incubation. Sensitivity and specificity analysis of results indicated that compared with the standard method (i.e., crystal violet tetrazolium agar), the method that performed the best, based on balanced accuracy (i.e., the average of sensitivity and specificity), was coliform Petrifilm evaluated for the presence of any growth after 48 h of incubation (sensitivity = 0.787; specificity = 0.839). This method can be easily adopted by the dairy industry as many processing facilities already test for coliforms using coliform Petrifilm. Improving the ability of processors to detect PPC will improve the quality of the fluid milk supply.

Hygiene assessments of school kitchens based on the microbiological status of served food.

According to European trends, more children eat at school canteens than ever before, therefore food safety and quality have become increasingly significant in recent years. Nevertheless, there are large differences in food safety levels in different school canteens. We hypothesize that the microbial status of the served meal represents on the general hygiene of the kitchen. Our research examines whether mesophilic aerobic bacteria measured in served food are connected with the level of hygiene in the catering unit, and whether this indicator can be used as a criterion for assessing school kitchens. Meal samples were collected from six school kitchens, and mesophilic aerobic bacterial count was measured. Samples were collected on five different days, so each kitchen was monitored five times. Two meals per visit were collected: a soup and a main course. Out of the 60 samples, 26 were good (CFU/g < 103), 24 were acceptable (CFU/g: 103&ndash;105), and in 10 samples, the microbial count was found to be above the limit (CFU/g > 105). Statistical calculations revealed that microbial contamination of served meals was influenced neither by the supplier nor by the type of meal (soup or main course). However, the level of hygiene in the serving kitchen significantly affects the microbial status of meals. Based on the results, a qualification system can be developed using the mesophilic aerobic bacterial count measurable in the served meal to assess hygiene. By regular determination of mesophilic aerobic bacterial count and the presence of Enterobacteriaceae, the food safety of a catering unit can be quantitatively evaluated.

Microbiological safety of flours used in follow up for infant formulas produced in Ouagadougou, Burkina Faso.

The prevalence of diarrheal diseases in children aged from 6 to 24 months in Burkina Faso is 38%. These diarrheas may be due to the consumption of contaminated weaning food. Therefore, the microbiological quality of follow up infant flours used as supplement foods is a key-point to reduce children diseases. In this study, the microbiological safety of locally-produced infant flours was investigated. One hundred and ninety-nine (199) samples were collected mainly in retails outlets and in Recovery and Nutrition Education Centers. According to the Burkina Faso regulations, microbiological analyses were carried out for Total Aerobic Mesophilic Flora (TAMF), thermotolerant coliforms, Salmonella spp. and yeasts/molds. The bacterial and fungal isolates were identified using phenotypic and genotypic methods and the study of the production of mycotoxins was carried out from the fungal isolates. In collected samples, the TAMF count ranged from 0 to 1.8 × 106 CFU/g with a total average of 6.3 × 104 CFU/g. About 2% of the samples had a microbial load exceeding the standards (105 CFU/g). No Salmonella spp. was isolated in the final infant flours. However, the presence of Enterobacteriaceae (Klebsiella spp. Enterobacter spp. and Cronobacter spp.) was detected and molecular characterization revealed also the presence of fungal species of the genus Aspergillus spp., Penicillium spp. and Fusarium spp. Some of these species were found to produce aflatoxins, ochratoxin A and fumonisins, which are potential carcinogenic toxins. These results demonstrated the need for a preventive approach based on the application of Hazard Analysis Critical Control Point in the food industry to ensure food safety of infant flours in Burkina Faso.

Effect of the lemon essential oils on the safety and sensory quality of salted sardines (Sardina pilchardus Walbaum 1792)

The main aim of this research was to investigate the biopreservative effects of lemon essential oil (EO) micro-emulsions on salted sardines. The experimental design included two experimental trials, SR1 and SR2 carried out with 25 ml of lemon EO micro-emulsion at 0.3 and 1.0% (v/v), respectively, and a control trial performed without EO addition. Chemical analyses on salted sardines inoculated with the EOs clearly showed a substantial persistence of several volatile organic compounds (VOCs) belonging to groups of monoterpene hydrocarbons, oxygenated monoterpenes and sesquiterpene hydrocarbons derived from EOs during the entire period of ripening. In particular, the molecules mostly represented were limonene, p-cymene and β-pinene. Immediately after the addition of EOs, the concentrations of all microbial groups decreased. The presence of Enterobacteriaceae, staphylococci and rod lactic acid bacteria (LAB) observed in the trials SR1 and SR2 was significantly lower than that registered for the control trial during the entire period of monitoring. Furthermore, the addition of EOs determined a lower accumulation of histamine in sardines compared to those of the control trial. The highest scores of sensory evaluation were registered for flavour and overall acceptability of the experimental trials in presence of EOs. On the basis of the increasing interest toward novel food preservatives, we conclude that the use of EOs to produce salted fishes represents a valid strategy to improve safety and sensory characteristics of salted sardines. This work has also economic implications, since the flavour improvement due to the addition of lemon EOs might increase the consumption of sardines by regular and new consumers.

Assessment of the bacterial community in directly brined Aloreña de Málaga table olive fermentations by metagenetic analysis

This study uses an “omics” approach to evaluate the bacterial biodiversity changes during fermentation process of natural green cracked Aloreña de Málaga table olives, from raw material to fermented fruit. For this purpose, two industries separated by almost 20km in Guadalhorce Valley (Málaga, Spain) were analysed for obtaining both brines and fruit samples at different moments of fermentation (0, 7, 30 and 120days). Physicochemical and microbial counts during fermentation showed the typical evolution of this type of processes, apparently dominated by yeasts. However, high-throughput barcoded pyrosequencing analysis of V2–V3 hypervariable region of the bacterial 16S rRNA gene showed at 97% identity the presence of 131 bacterial genera included in 357 operational taxonomic units, not detected by the conventional approach. The bacterial biodiversity was clearly higher in the olives at the moment of reception in the industry and during the first days of fermentation, while decreased considerably as elapse the fermentation process. The presence of Enterobacteriaceae and Lactobacillaceae species was scarce during the four months of study. On the contrary, the most important genus at the end of fermentation was Celerinatantimonas in both brine (95.3% of frequency) and fruit (89.4%) samples, while the presence of well-known spoilage microorganisms (Pseudomonas and Propionibacterium) and halophilic bacteria (Modestobacter, Rhodovibrio, Salinibacter) was also common during the course of fermentation. Among the most important bacterial pathogens related to food, only Staphylococcus genus was found at low frequencies (<0.02% of total sequences). Results show the need of this type of studies to enhance our knowledge of the microbiology of table olive fermentations. It is also necessary to determine the role played by these species not previously detected in table olives on the quality and safety of this fermented vegetable.

Effect of slaughterhouse and day of sample on the probability of a pig carcass being Salmonella-positive according to the Enterobacteriaceae count in the largest Brazilian pork production region

Sources of contamination of carcasses during slaughter include infected pigs as well as environmentally related sources. There are many microbial indicators that can be used in the processing of food to assess food hygiene and the safety of food processing. The presence of some microbial indicators can be viewed as a result of direct or indirect contamination of a food with fecal material. The presence of Enterobacteriaceae is often used as a hygiene indicator, as they are found both in the environment and in the intestine of warm-blooded animals. An association between Salmonella isolation and Enterobacteriaceae count (EC) on pre-chill carcasses has been described, however the impact of slaughterhouse and the day of sampling on the occurrence of Salmonella has not been previously investigated. To this end, mixed logistic regressions (MLRs) with random effects and fixed slopes were performed to assess the change in EC and its correlation with Salmonella occurrence using two data sets. The first describes the EC and Salmonella isolation in 60 pork carcasses in one slaughterhouse sampled at 11 different slaughter steps, including the carcass as a random effect. The second describes the EC and Salmonella isolation on 1150 pre-chill carcasses sampled in 13 slaughterhouses over 230 sampling days, and the model combined two random intercepts, slaughterhouse and date of sampling nested with slaughterhouse (day/slaughterhouse). Statistically significant associations (p<0.0001) between the log of the EC and Salmonella occurrence were found in all models. Nevertheless, although a strong association was found between Enterobacteriaceae and Salmonella contamination in pork carcasses, this association was not constant, given that there was a high variation in the probability of a carcass being positive for Salmonella according to the EC mainly between days of samples. The effect of the day of sampling on Salmonella prevalence was so large that the predictive value of the EC count for Salmonella isolation on a daily basis was compromised. It is possible that on some days batches with a high prevalence of Salmonella carriers shedding a high number of Salmonella were slaughtered. On these days, the potential for contamination/cross-contamination of carcasses will be so large that even hygienic slaughter, confirmed by the low EC on carcasses, will not be able to prevent the presence of Salmonella on some carcasses. The results of this study demonstrate that, despite the statistically significant association found, it may be difficult to predict when hygiene failure measured via EC actually indicates Salmonella contamination, and neither the inverse.

Microbiological Quality of Raw Dried Pasta from the German Market, with Special Emphasis on Cronobacter Species

The microbiological quality of 132 dried pasta products available on the German market, originating from 11 different countries, was studied. Sample materials included soft or durum wheat products, some of which produced with other ingredients such as eggs, spices, or vegetables. Parameters included hygiene indicators (aerobic plate count, mold count, the presence of Enterobacteriaceae) and pathogenic/toxinogenic bacterial species (Salmonella spp., Staphylococcus aureus, presumptive Bacillus cereus, and Cronobacter spp.). The overall results of hygiene parameters indicated a satisfactory quality. Salmonella was not found in any sample. Three samples were positive for S. aureus (10(2) to 10(4) colony forming unit (CFU)/g). Presumptive B. cereus at levels of 10(3) to 10(4) CFU/g were detected in 3 samples. Cronobacter spp. were isolated from 14 (10.6%) products. Of these, 9 isolates were identified as C. sakazakii, 2 each as C. turicensis and C. malonaticus, and 1 as C. muytjensii. The isolates were assigned to 9 multilocus sequence typing (MLST) sequence types and to 14 different PFGE profiles. Although pasta products are typically cooked before consumption, some consumers, and children in particular, may also eat raw pasta as nibbles. Raw pasta seems to be a relevant source of exposure to dietary Cronobacter spp., although health risks are probably restricted to vulnerable consumers. High numbers of presumptive B. cereus as found in some samples may be a risk after improper storage of cooked pasta products because toxinogenic strains are frequently found within this species.

Analysis of microbiota in stable patients with chronic obstructive pulmonary disease.

To identify the bacterial diversity (microbiota) in expectorated sputum, a pyrosequencing method that investigates complex microbial communities of expectorated sputum was done in 19 stable chronic obstructive pulmonary disease patients (mean (SD) FEV1: 47 (18%) of predicted value). Using conventional culture, 3 phyla and 20 bacterial genera were identified, whereas the pyrosequencing approach detected 9 phyla and 43 genera (p < 0.001). In sputum the prevalent genera with pyrosequencing approach were Streptococcus, Actinomyces, Neisseria, Haemophilus, Rothia, Fusobacterium, Gemella, Granulicatella, Porphyromonas, Prevotella and Veillonella. Enterobacteriaceae, detected frequently in conventional culture, were not significantly detected with pyrosequencing methods. In addition, we found that important pathogens such as Haemophilus and Moraxella were detected more frequently with the new genetic procedures. The presence of Enterobacteriaceae is probably overestimated with conventional culture, whereas other difficult cultivable pathogens are underestimated. These studies open a new perspective for evaluating the role of bacterial colonization in chronic obstructive pulmonary disease pathogenesis and progression.

Physicochemical evaluation, nutraceutical composition and HPLC‑UV fingerprint of Helicanthus elastica (Desr.) Danser (Indian Mango Mistletoe)

Background: Helicanthus elastica (Desr.) Danser (Loranthaceae) is less‑known Indian medicinal mistletoe growing commonly on mango trees as hemiparsites. It is used to prevent abortion, in vescical calculi and kidney affections. These groups of plants are medicinally important as they are potential sources of anticancer, immunomodulatory, hepatoprotective, antimicrobial and antioxidant molecules. Materials and Methods: In the current study whole plant of H. elastica growing on mango trees is collected and subjected for physicochemical and nutraceutical analysis aiding standard methodology. The total ethanolic extract was fingerprinted with HPLC‑UV. Results: Parameters like moisture content, total ash, water‑soluble ash, acid‑insoluble ash, alcohol‑soluble and water‑soluble extractive, successive extractive values by cold and hot extraction, heavy metals like arsenic, lead, cadmium and mercury, total bacterial count, total fungal count, presence of enterobacteriaceae, Escherichia coli, Salmonella spp., Pseudomonas aeruginosa and Staphylococcus aureus were tested under the head physicochemical examination. Determination of carbohydrate, fat, protein and fiber, calorific value, vitamins like niacinamide, pyridoxine, riboflavin, thiamine and ascorbic acid, trace elements like iron and zinc were estimated. Conclusion: The physicochemical tests are used as parameters for routine analysis and quality control of the plant. The investigations revealed appreciable quantity of important vitamins and trace elements in the plant. The HPLC‑UV fingerprint would be an efficient tool for the standardisation and quality control of the mistletoe extract. Key words: Helicanthus elastica, hemiparsites, HPLC-UV fingerprint, mango mistletoe, nutraceutical composition