Combined Vertical-Horizontal Flow Biochar Filter for Onsite Wastewater Treatment—Removal of Organic Matter, Nitrogen and Pathogens

This study investigated the performance of a combined vertical-horizontal flow biochar filter (VFF-HFF) system in terms of organic matter, total nitrogen (Tot-N), Escherichia coli and Salmonella removal and explored the effects of hydraulic loading rate (HLR) on pollutant removal. The combined VFF-HFF system used biochar as the filter medium and comprised two stacked sections: (i) an aerobic vertical flow filter (VFF) in which the wastewater percolated through the biochar medium in unsaturated mode and (ii) a horizontal flow filter (HFF), in which the biochar was saturated with water and had limited access to air, to enable anaerobic conditions and enhance the denitrification process. The system was tested over 126 weeks using real wastewater applied at different HLR (23, 31, 39 L m−2 day−1). The results showed that long-term removal of organic matter in the entire system was 93 ± 3%, with most (87 ± 5%) occurring in the VFF. For Tot-N, the long-term removal was 71 ± 12%, with increasing trends for nitrification in the VFF and denitrification in the HFF. Mean long-term nitrification efficiency in the VFF was 65 ± 15% and mean long-term denitrification efficiency in the HFF 49 ± 14%. Increasing HLR from 23 to 31 L m−2 day−1 increased the nitrification efficiency from 42 to 61%. Increasing the HLR further to 39 L m−2 day−1 decreased the denitrification efficiency from 45 to 25%. HLR had no significant effects on VFF and HFF performance in terms of E. coli and Salmonella removal, although the VFF achieved a 1.09–2.1 log10 unit reduction and the HFF achieved a 2.48–3.39 log10 unit reduction. Thus, long-term performance, i.e., removal of pollutants measured during the last 52 weeks of the experiment, was satisfactory in terms of organic matter and nitrogen removal, with no signs of clogging, indicating good robustness of the combined VFF-HFF biochar filter system.

Saliva and Serum Immune Responses in Apical Periodontitis

Apical periodontitis is an inflammatory reaction at the apex of an infected tooth. Its microbiota resembles that of marginal periodontitis and may induce local and systemic antibodies binding to bacteria- and host-derived epitopes. Our aim was to investigate the features of the adaptive immune response in apical periodontitis. The present Parogene cohort (n = 453) comprises patients with cardiac symptoms. Clinical and radiographic oral examination was performed to diagnose apical and marginal periodontitis. A three-category endodontic lesion score was designed. Antibodies binding to the bacteria- and host-derived epitopes were determined from saliva and serum, and bacterial compositions were examined from saliva and subgingival samples. The significant ORs (95% CI) for the highest endodontic scores were observed for saliva IgA and IgG to bacterial antigens (2.90 (1.01–8.33) and 4.91 (2.48–9.71)/log10 unit), saliva cross-reacting IgG (2.10 (1.48–2.97)), serum IgG to bacterial antigens (4.66 (1.22–10.1)), and Gram-negative subgingival species (1.98 (1.16–3.37)). In a subgroup without marginal periodontitis, only saliva IgG against bacterial antigens associated with untreated apical periodontitis (4.77 (1.05–21.7)). Apical periodontitis associates with versatile adaptive immune responses against both bacterial- and host-derived epitopes independently of marginal periodontitis. Saliva immunoglobulins could be useful biomarkers of oral infections including apical periodontitis—a putative risk factor for systemic diseases.

Effect of zero-valent iron amendment on the performance of biosand filters

The study compared the performance of a biosand filter (BSF) with two BSFs modified by introducing a layer of zero-valent iron (ZVI) during a long-duration test (4 months) mimicking the household use pattern in developing countries. Results of the study showed that for bacterial removal, ZVI-amended BSFs outperformed the BSF by at least 1 log10 unit throughout the filter operation. Effluent turbidity in the BSF and modified BSFs was not significantly influenced by influent values for the turbidity range tested in the study (17.0–45.4 NTU). Removal efficiency of nitrates was higher in the modified BSFs, with up to ∼89% removal in the modified BSFs compared with ∼29% in the BSF. Sharp decline in dissolved oxygen (DO) was observed during the passage of water through the filters. The DO decline was more in the modified filters compared with the conventional BSF. Effluent iron remained within the drinking-water quality standards. The study thus indicates the potential of ZVI to improve the performance of BSFs.

Estimating Generalised Flood Sew Coeficients for Peninsular Malaysia

Flood estimations based on itting the frequency of occurrence of annual peak discharges using the Log-Pearson Type 3 distribution are commonly used but they are sensitive to the skew coeficients of the gauging stations. The estimation accuracy can be improved by using a weighted average population skew coeficient calculated from the sample station skew and the generalised unbiased skew. The U.S. Water Resources Council (WRC) has documented guidelines for estimating the generalised skew coeficients and published a map of generalised skew values for the United States. The map shows isolines of skew coeficient values and the average skew coeficient for each 1-degree quadrangle of latitude and longitude for the United States. Following the WRC guidelines, many of the state authorities in the US have developed the generalised skew coeficients separately on a state/regional basis. In Malaysia, the Log Pearson Type 3 distribution has been widely used for lood peakestimation but there are no guidelines available for estimating the generalised skew coeficients for use in con–unction with the distribution and as more accurate results can be obtained if these data are available, it is clear that a regional lood skew study is needed. With the regional sew data available, the peak low can be simply and easily calculated with the aid of a software such as HEC-SSP. The aim of this paper is to use the WRC guidelines to derive the generalised skew coeficients using the peakannual discharge data of Peninsular Malaysia for general use.The WRC recommended several techniques for estimating and evaluating generalised s—ew of the Log-Pearson Type 3 distribution for the annual peakdischarges. Station skews (skew coeficients computed from gauging station records) and unbiased and weighted skews derived from these station skews are to be used to develop these techniques. In this study, peak discharge records at 66 gauging stations having 16 or more annual peak discharges in Peninsular Malaysia were selected for computing station skews. Station skew values ranged from -0.831(log10 unit) to 1.475 (log10 unit).The three techniques recommended by WRC used for estimating the generalised skew of annual peak discharges were adopted for this study. These methods are: (1) An isoline map, (2) a prediction equation (3) a regional mean skew. Attempts to develop a prediction equation were unsuccessful. An error analysis showed that the regional mean skew method has a lower MSE (mean square error) than that obtained from the state wide generalised skew coeficient contour map. As a result, the mean station skew for the selected gauging stations can be used to estimate the generalised skew for any gauging site in the peninsula.The mean skew is -0.022 (log10 unit) and the associated mean square error is 0.05 (log10 unit).

Bactericidal effect of photodynamic therapy against methicillin-resistant Staphylococcus aureus strain with the use of various porphyrin photosensitizers.

Photodynamic therapy (PDT) is based on photosensitizers activated by light of appropriate wavelength. Their activation leads to generation of singlet oxygen and free radicals responsible for the cytotoxic effect. The aim of this project was to compare the bactericidal effect of PDT using different porphyrin photosensitizers against a methicillin-resistant Staphylococcus aureus strain. Exogenous sensitizers (protoporphyrin IX and newly synthesized derivative, protoporphyrin diarginate) induced a 3 log10-unit reduction in bacterial viable counts. With the use of endogenous, ALA-induced porphyrins, a 1.6 log10-unit reduction was obtained. The sensitizers tested executed their antibacterial activity with no essential change in the antibiotic resistance pattern of the studied strain.

Survival of Escherichia coli O157:H7 in Full- and Reduced-Fat Pepperoni after Manufacture of Sticks, Storage of Slices at 4°C or 21°C under Air and Vacuum, and Baking of Slices on Frozen Pizza at 135, 191 and 246°C

Pepperoni batter was prepared with fat contents of about 15, 20, and 32% (wt/wt) and inoculated with a pediococcal starter culture and ≥2.0 × 107 CFU/g of a five-strain inoculum of Escherichia coli O157:H7. The batter was fermented at 96°F (ca. 36°C) and 85% relative humidity (RH) to pH ≤ 4.8 and then dried at 55°F (ca. 13°C) and 65% RH to a moisture/protein ratio of ≤1.6:1. For storage, slices were packaged under air or vacuum and stored at 39°F (ca. 4°C) and 70°F (ca. 21°C). For baking, frozen slices were placed on retail frozen cheese pizzas that were subsequently baked at 275°F (ca. 135°C), 375°F (ca. 191°C), or 475°F (ca. 246°C) for 0 to 20 min. Appreciable differences related to fat levels were observed after drying; pathogen numbers decreased by 1.04, 1.31 and 1.62 log10 units in sticks prepared from batter at initial fat levels of 15, 20, and 32%, respectively. During storage, the temperature rather than the atmosphere had the greater effect on pathogen numbers, with similar viability observed among the three fat levels tested. At 70°F (ca. 21°C), compared to original levels, pathogen numbers decreased by ≥5.56 and ≥4.53 log10 units within 14 days in slices stored under air and vacuum, respectively, whereas at 39°F (ca. 4°C) numbers decreased by ≤2.43 log10 CFU/g after 60 days of storage under either atmosphere. Baking, as expected, resulted in greater reductions in pathogen numbers as the temperature and/or time of baking increased. However, it was still possible to recover the pathogen by enrichment after baking frozen slices on frozen pizza at 475°F (ca. 246°C) for 10 min or at 375°F (ca. 191°C) for 15 min. The calculated D values for all three temperatures tested increased as the fat content of the batter increased from 15 to 20 to 32%. The present study confirmed that fermentation and drying were sufficient to reduce levels of E. coli O157:H7 in pepperoni sticks by <2.0 log10 CFU/g. Storage of slices for at least 14 days at ambient temperature under air resulted in a >5.5-log10-unit total reduction of the pathogen. Baking slices on frozen pizza for at least 15 min at 475°F (ca. 246°C) or 20 min at 375°F (ca. 191°C) was necessary to reduce pathogen numbers to below detection by both direct plating and enrichment.

Viability of Escherichia coli O157:H7 in Salami Following Conditioning of Batter, Fermentation and Drying of Sticks, and Storage of Slices

The fate of Escherichia coli O157:H7 was monitored in salami during conditioning of batter, fermentation and drying of sticks, and storage of slices. The raw batter (75% pork:25% beef, wt/wt, fat content about 20%) was inoculated with a pediococcal starter culture (about 108 CFU/g) and a five-strain cocktail of E. coli O157:H7 (≥2 × 107 CFU/g) and stuffed into 104-mm diameter fibrous casings. After being refrigerated at 4°C or being tempered at 13°C, frozen at −20°C, and thawed at 4°C, or being frozen at −20°C, and thawed at 4°C, the inoculated batter was fermented at 24°C and 90% relative humidity (RH) to pH ≤4.8, dried at 13°C and 65% RH to a moisture/protein ratio of ≤1.9:1, and then stored at 4 or 21°C under air or vacuum. For salami sticks sampled immediately after drying, appreciable differences were evident among the various batter-conditioning treatments; pathogen numbers were reduced from original levels by 2.1, 1.6, or 1.1 log10 units when batter was tempered, frozen, and thawed, frozen and thawed, or refrigerated, respectively. Similarly, regardless of storage temperature or atmosphere, within 7 days salami slices cut from sticks prepared from batter that was tempered, frozen, and thawed (2.7- to 4.9-log10-unit reduction) or frozen and thawed (2.3- to 4.8-log10-unit reduction) displayed a greater impact on pathogen numbers than slices cut from sticks prepared from batter that was refrigerated (1.6- to 3.1-log10-unit reduction). The effects of batter conditioning notwithstanding, a greater reduction in levels of E. coli O157:H7 was observed when slices were stored at 21°C compared to otherwise similar slices stored at 4°C. After storage for 60 days the pathogen was only detected by enrichment in slices stored at 21°C, whereas pathogen levels ranged from 1.4 to 4.5 log10 CFU/g in slices stored at 4°C. Differences related to storage atmosphere were first observed after slices were stored for 21 days. Such differences were more readily demonstrable after 60 and 90 days, with pathogen numbers for treatments that were statistically different ranging from 0.6- to 1.5-log10 units higher on slices stored under vacuum than in air. These data emphasize the need to implement multiple barriers to appreciably reduce numbers of E. coli O157:H7 in salami.

Survival of Escherichia coli O157:H7 during the Manufacture of Pepperoni†‡

This study investigated the growth and survival of Escherichia coli O157:H7 during the manufacture of pepperoni to determine whether a 5-log10-unit decline in numbers, as recommended by the U.S. Food Safety and Inspection Service (FSIS), could be achieved. A range of pepperoni formulations with variations in salt (2.5 to 4.8%) and sodium nitrite (100 to 400 ppm) levels, and with pH (4.4 to 5.6) adjusted by manipulation of dextrose concentrations were prepared. The batters produced were inoculated with E. coli O157:H7 380-94 at a level of approximately 6.70 log10 CFU/g; changes in pathogen numbers, pH, titratable acidity, and sodium nitrite concentrations were monitored during fermentation and drying. With the standard commercial formulation (i.e., 2.5% salt, 100 ppm sodium nitrite, pH 4.8) E. coli O157:H7 numbers declined by approximately 0.41 log10 CFU/g during fermentation and a further 0.43 log10 CFU/g during subsequent drying (7 days). A regression equation was fitted to the data which showed significantly (P < 0.001) greater reductions in pathogen numbers in samples with increased salt and sodium nitrite contents and lowered pH. However declines were in all cases less than the target reduction of 5 log10 CFU/g.

Listeria monocytogenes Occurrence and Growth at Refrigeration Temperatures in Fresh Blue Crab (Callinectes sapidus) Meat

In this study, 126 samples of freshly cooked and picked blue crab (Callinectes sapidus) meat collected from different processing facilities were analyzed for the presence of Listeria spp. Thirteen samples (10%) were positive for Listeria, with 10 samples positive for L. monocytogenes and 3 samples positive for L. innocua. Fraser broth was used in a 5-tube most probable number (MPN) enumeration, in duplicate, of Listeria in 25-g samples incubated at 36°C for 24 h and plated in modified Oxford agar and blood agar with API strip confirmation. The levels of Listeria cells found in fresh blue crab meat were always less than 100/g with only one exception, in which the MPN index was 1,100/g. A L. monocytogenes strain (168) isolated from fresh blue crab meat was inoculated (less than 50 CFU/g) into pasteurized crab meat and incubated at 1.1, 2.2, and 5°C for 21 days. Growth curves were obtained by analyzing 2-g samples at intervals of 0, 8, 10, 12, 14, 16, 19, and 21 days. When inoculated into pasteurized blue crab meat, L. monocytogenes had an increased growth rate as the storage temperature increased, with approximately a 7-log10-unit increase in population at 5°C and only a 2.5-log10-unit increase in population at 1.1 °C after the 21 days of incubation.

Susceptibility of Preevisceration Washed Beef Carcasses to Contamination by Escherichia coli O157:H7 and Salmonellae†

Prerigor bovine cutaneous truncii muscle was subjected to a simulated preevisceration wash procedure 10 min after the hide was removed from the carcass. Five minutes after washing, the preevisceration washed tissue and unwashed control tissues were contaminated with a direct application of either fresh bovine manure or bovine manure which had been inoculated with five-strain mixtures of either Escherichia coli O157:H7 or salmonellae to simulate fecal contamination on a carcass. The manure which was inoculated with the bacteria had been previously irradiated to eliminate enterobacteriaceae. The contamination was allowed to adhere to the tissues for 10 min and then washed off with distilled water. Surface free energy of the preevisceration washed tissue was significantly lower than that of tissue which had not been subjected to preevisceration washing. Scanning electron micrographs of the tissue samples revealed plant material adhering to the surface of the control tissues, but not to the preevisceration washed tissues. Microbial populations of both total aerobic bacteria and enterobacteriaceae were approximately 0.7 log10 unit greater on the control tissue, in comparison to the preevisceration washed tissue. Preevisceration washing may be beneficial in reducing the susceptibility of animal carcasses to further contamination.