Quality assessment of Mediterranean horse mackerel (Trachurus mediterraneus) and blue jack mackerel (Trachurus picturatus) during storage in ice

Sensory, physicochemical and microbiological analyses were carried out on Mediterranean horse mackerel and blue jack mackerel in order to determine quality changes during storage in ice. Sensory results indicated that whole Mediterranean horse mackerel and blue jack mackerel used in this trial had a shelf-life of 10 and 7 days, respectively. Fishtester and Fishchecker measurements showed that the quality of blue jack mackerel had deteriorated more rapidly. Fishtester values of the blue jack mackerel and the Mediterranean horse mackerel were reduced from 59 ± 10.1 to 14.8 ± 2.8 and from 85.3 ± 13.2 to 25.6 ± 7.8, respectively during the storage time. Fishchecker gave red sign (unfit for consumption) for first time on day 7 and day 6 in Mediterranean horse mackerel and blue jack mackerel, respectively. The pH of Mediterranean horse mackerel muscle was consistently lower than the pH of blue jack mackerel muscle during the storage period. The pH values of both species muscle increased gradually with time and pH value equal to 7 had almost coincided with the end of shelf-life. Bacterial loads in both fish species muscle remained rather low prior to day 10 (<6 log₁₀ cfu/g). The longer shelf-life of Mediterranean horse mackerel as determined by sensory evaluation was not reflected in microbiological results suggesting that the difference was due to lower rate of autolysis rather than to reduced microbial activity.     

Some hazards associated with the production of a popular roasted meat (tsire) in Zaria,Nigeria

Some hazards associated with the entire production process of tsire (a local kebab) were identified in three production centres. Microbiological analyses showed tsire to have highest aerobic plate count of log₁₀ 6.27; B. cereus count was highest at log₁₀ 3.30 cfu/g; Clostridium perfringens count was highest at log₁₀ 2.92 cfu/g; Staphylococcal count was highest at log₁₀ 3.96 cfu/g; Coliform count was highest at log₁₀ 4.08 cfu/g; yeast and mould count was highest at log₁₀ 2.49 cfu/g. The proximate analysis showed tsire to averagely have 11.87% moisture, 31.77% protein, 23.16% fat and 2.43% salt. The critical appraisal of the production process indicated potential hazards in the raw meat, environmental contamination as well as post-process handling contamination from humans and the environment. The nature of microorganisms associated with tsire production as shown by this study calls for worry from the public health standpoint. In the light of this, efforts should be made by public health services with regard to improving its production in order to reduce the associated hazards.     

Microbiological changes in farm reared freshwater prawn (Macrobrachium rosenbergii de Man) in ice

The microbiological changes in farm reared freshwater prawn (Macrobrachium rosenbergii de Man) during ice storage were studied. A total of 156 bacterial cultures from fresh and ice-stored farmed freshwater prawn were isolated and characterized. Total aerobic, mesophilic and psychrotrophic counts and hydrogen sulphide producing bacterial counts were determined. The total aerobic counts at 20 and 37 °C on fresh prawn was in the range of 4–5 log₁₀ cfu g⁻¹. Aerobic counts on M. rosenbergii at 20 °C and 7 °C exceeded 10⁷ cfu g⁻¹ by the end of storage, of which 40–52% were H₂S producers. Gram-negative bacteria constituted 73% of the total flora of fresh prawn and Enterobacteriaceae and Aeromonadaceae dominated. After 19 days of iced storage, more than 80% of the bacterial flora of prawn were Gram-negative. Pseudomonas, Aeromonas hydrophila, A. veronii boivar sobria and Shewanella putrefaciens were identified as the dominant spoilage organisms of farm reared M. rosenbergii stored in ice. This study confirms that freshwater prawn carry significant numbers of motile aeromonads capable of growth at low temperature. The results of the study indicated that the shelf-life of freshwater prawn as determined by microbiological data is 12–16 days. Immediate icing of harvested M. rosenbergii is essential to improve the microbiological stability.     

Microbial profile of buffalo sausage during processing and storage

A study was made on the microbial levels of buffalo sausage during preparation and storage at 4 ± 1 °C. Microbial counts in raw minced meat were, total plate count (TPC) (log cfu/g) 5.41 ± 0.25; coliforms (MPN/g) 23.2; Staphylococcus aureus (log cfu/g) 1.57 ± 0.11; yeasts and molds (log cfu/g) 2.29 ± 0.07 and lactic acid bacteria (LAB) (log cfu/g) 0.60 ± 0.20. Sausage emulsion showed similar trend in microbial counts with minimal microbial contamination during the preparation of emulsion. Cooked buffalo sausage gave the following microbial counts: TPC (log cfu/g) 3.75 ± 0.31; coliforms (MPN/g) 0.2; LAB (log cfu/g) 0.07 ± 0.01; yeast and molds (log cfu/g) 0.72 ± 0.07. S. aureus, Clostridium perfringens and Bacillus cereus were not detected in cooked sausages. These results indicate that steam cooking for 45 min followed in the study was effective in reducing the microbial counts substantially. The investigation revealed that shelf life of cooked buffalo sausage was 31 days in either vacuum or CO₂ at 4 ± 1 °C. The results indicated that spoilage of vacuum packed cooked buffalo sausage was likely due to LAB while microflora other than LAB may be responsible for spoilage of CO₂ packed cooked buffalo sausage. The study suggests that measures such as low initial microbial counts, hygienic precautions during preparation of sausage, steam cooking for 45 min, vacuum or CO₂ packing and storage at 4 ± 1 °C would control the microbial growth and provide wholesomeness and safety to the buffalo sausage.     

Effect of washing and disinfecting containers on the microbiological quality of fresh milk sold in Bamako (Mali)

The present study aimed to improve the microbiological quality of the milk, from the cow’s udder to the selling point by container washing and disinfecting. The total counts (TC), Enterobacteriaceae counts (EBC) were used as quality indicators.The results showed a significant decrease of milk flora at the selling point, from 1.6 × 10⁷ ± 1.3 × 10⁷ (control) to 4.8 × 10⁵ ± 0,8 × 10⁷ (intervention) colony forming units per ml (cfu ml⁻¹) for TC and 1.2 × 10⁶ ± 1.1 × 10⁶ (control) to 0.9 × 10⁴ ± 1.1 × 10⁴ (intervention) cfu ml⁻¹ for EBC. The study suggests that in milk production area, besides udder infection and water quality, hygiene behaviour with respect to hand washing, container’s cleaning and disinfection are the key areas that remain of relevance to milk hygiene intervention.     

Effect of gamma irradiation on Listeria ivanovii inoculated to iceberg lettuce stored at cold temperature

A low-dose gamma irradiation considerably reduced the total plate counts, psychrotrophic bacteria, lactic acid bacteria, and inoculated Listeria ivanovii on shredded iceberg lettuce. The total plate count of the lettuce irradiated at 1.0 kGy was reduced by 3.38 log₁₀ cfu/g on 0 storage day and to below the limit of detection (<2 log₁₀ cfu/g) as the cold storage was extended. Irradiation at 0.5 kGy effectively reduced the psychrotrophic bacterial counts and lactic acid bacterial counts in the lettuce to below the limit of detection (2 log₁₀ cfu/g). Irradiation at 1.0 kGy reduced L. ivanovii inoculated onto the shredded iceberg lettuce to below the limit of detection. The results showed that an irradiation at 1 kGy eliminates the bacterial contamination from the lettuce sample without any sensorial quality defect.     

The incidence of significant foodborne pathogens in domestic refrigerators

The interior surfaces of household refrigerators are at risk of becoming contaminated with foodborne pathogens, increasing the risks of cross-contamination to other food items, including higher risk ready-to-eat foods. This study determined the incidence of a number of significant foodborne pathogens, and the general hygienic status (as estimated by total viable counts (TVCs), and total coliform counts (TCCs)) on the interior surfaces of domestic refrigerators (n = 342). Campylobacter spp., Salmonella spp. and Escherichia coli O157:H7 were not recovered from any refrigerators, but Staphylococcus aureus was recovered from 6.4%, Listeria monocytogenes and E. coli from 1.2% and Yersinia enterocolitica from 0.6% of examined refrigerators. As the recovered species can survive and grow under refrigeration or conditions of mild temperature abuse, such pathogens may transfer to (and develop to clinically significant numbers in) food in domestic fridges. Such risks are of particular concern in relation to “ready-to-eat” foods, which will not receive further bactericidal treatments (cooking) before consumption. The study estimated TVCs ranging from 2.91 log₁₀ cfu/cm² to 8.78 log₁₀ cfu/cm² and TTCs ranging from 0.045 log₁₀ cfu/cm² to 5.96 log₁₀ cfu/cm² indicating very poor standards of consumer refrigerator management and hygiene, and posing risks to consumer health. The study findings highlight the importance of adequate temperature control and thorough, regular cleaning of domestic refrigerators to ensure food safety, and of effective cooking as the last link in the domestic food service chain.     

Microbiology of retail mung bean sprouts vended in public markets of National Capital Region,Philippines

This study is the first attempt in the Philippines to conduct a region-wide assessment of the microbiological quality of retailed mung bean sprouts. Production and vending practices of selected stakeholders were also determined. Ninety-four percent of the samples tested positive for the presence of Salmonella spp. and some samples had Coliform and Escherichia coli counts as high as 5.90 and 5.50 log₁₀ CFU · g⁻¹, respectively. The TPC, YMC and LAB were established to be as high as 11.38, 5.90 and 10.47 log₁₀CFU · g⁻¹, respectively. The poor microbiological quality of most of the tested sprouts was attributed to unhygienic sprout production and retailing practices. To improve the microbiological quality of the retailed sprouts, adherence to sprouting industry best practices is strongly recommended.     

Microbiological evaluation of fresh-cut organic vegetables produced in Zambia

This study was undertaken to assess the microbiological quality of fresh-cut organic vegetables produced in Zambia. Fresh-cut organic mixed vegetables and green beans produced in Zambia were analysed for aerobic plate counts, coliforms, Enterobacteriaceae, Escherichia coli, Bacillus cereus, Clostridium perfringens, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and yeast and mould counts. The study included 160 samples for most of the parameters. The vegetables were grown on farms meant primarily for the export market. The vegetables were treated/washed with 150 μg ml⁻¹ chlorine solution at the processing plant prior to sampling. The aerobic plate count ranged between 3 log₁₀ and 9.7 log₁₀ CFU/g, with the highest count recorded for green beans. The largest grouping (26.1%) of vegetable samples fell between 3 and 4 log₁₀ CFU/g. Coliform counts were between 1.0 log₁₀ and 7.7 log₁₀ CFU/g. The highest incidence level was 31.4% for total coliform counts between 3 log₁₀ and 4 log₁₀ CFU/g. E. coli was only detected on mixed vegetables in the range of 0.6 log₁₀ to 3 log₁₀ CFU/g, while Enterobacteriaceae counts ranged between 1.6 log₁₀ and 9.8 log₁₀ CFU/g with the highest counts being found on green beans. The highest incidence level was of 25.8% for counts within the same range as the aerobic plate counts. Yeast and mould counts showed the highest incidence level between 5 log₁₀ and 6 log₁₀ CFU/g with an overall range between 1.5 log₁₀ and 5.6 log₁₀ CFU/g. L. monocytogenes, Salmonella spp. and S. aureus were detected in 20%, 23.1% and 83.9% of samples, respectively . C. perfringens and B. cereus were not detected in any of the samples analysed.     

Pre-processed bovine milk quality in Trinidad: Prevalence and characteristics of bacterial pathogens and occurrence of antimicrobial residues in milk from collection centres

The study was conducted to assess the impact of the changes in the milk collection system in Trinidad (from twice daily collection to once, introduction of chilling facilities to the collection centres and transportation of milk to the processing plant in insulated truck instead of in metal churns at ambient temperature) on the microbial load and antimicrobial residue quality of the milk as well as the temperature and pH of milk, using standard methods. The presence of antimicrobial residues was detected using the Delvo test kit. Of a total of 266 milk samples from churns, the mean ± sd temperature and log₁₀ ± sd TAPC per ml was 20.36 ± 7.91 °C and 6.3 ± 1.09 respectively. For 20 milk samples from the chillers, the mean temperature and log₁₀ ± sd TAPC per ml was 15.10 ± 2.73 °C and 7.04 ± 0.33 respectively compared with corresponding values for 36 samples collected from the truck, 11.64 ± 4.22 °C and 7.11 ± 0.62 respectively (P < 0.05; X²). The mean TAPC, staphylococcal and E. coli counts per ml of milk from churns were significantly (P < 0.05; X²) higher for milk at low temperature (0–20 °C) compared with milk at high temperature (>30 °C). Eight (4.2%) of 192 milk samples tested contained antimicrobial residues. Of 168 S. aureus isolates tested, 24 (14.3%) were enterotoxigenic while 53 (45.3%) of 117 isolates tested exhibited resistance to various antimicrobial agents while of 386 isolates of E. coli tested, 3 (0.8%) were O157 strain and 129 (64.5%) of 200 isolates exhibited resistance to antimicrobial agents. It was concluded that despite the changes, the microbial load of milk was still high suggesting poor sanitary practices at the farm level. The detection of antimicrobial residues agents coupled with toxigenic S. aureus and E. coli isolates could pose health hazards to consumers.     

Microflora of traditional starter made from cassava for “attiéké” production in Dabou (Côte d’Ivoire)

Attiéké is a food made from cassava in Côte d’Ivoire by fermentation. The process uses a traditional starter. Studies on 81 starter samples from 3 villages showed that the dominant microflora consists of lactic acid bacteria (5.7 × 10⁷ cfu/g), yeasts (5.5 × 10⁷ cfu/g), Bacillus (3.8 × 10⁷ cfu/g), Enterococcus (3.0 × 10⁶ cfu/g), total coliforms (3.0 × 10⁶ cfu/g), thermotolerant coliforms (8.0 × 10³ cfu/g) and mould (2.0 × 10⁶ cfu/g). Lactic acid bacteria, Bacillus spp, yeasts, faecal Enterococci and moulds are organisms which could play a role in the cassava fermentation. Coliforms may indicate contamination from the environment during production.     

Exposure assessment for Bacillus cereus in ready-to-eat Kimbab selling at stores

Kimbab, which is rolled cooked-rice and other foodstuffs in dried laver, is a representative ready-to-eat (RTE) food in Korea. Bacillus cereus is a significant hazard associated with Kimbab. However, the knowledge of the dose–response relationship for B. cereus is limited, now it is not possible to conduct a full microbial risk assessment. Therefore, our study may be focused on the exposure assessment from retail store to the point of consumption in a Kimbab contaminated with B. cereus. The survey data for preparing and selling of Kimbab, predictive growth model, and consumer eating patterns were combined with probabilistic modeling to simulate the level of B. cereus in store Kimbab at the time of consumption. The estimated contamination levels ranged from as minimum (5% percentile) −3.63 log cfu/g, median (50% percentile) 1.39 log cfu/g, mean 1.57 log cfu/g, and a maximum (95% percentile) 7.31 log cfu/g. However, there is always a paucity of data to conduct such full exposure assessments with associated variability and uncertainty, coupled with a validation step. Therefore, we suggest that additional studies to allow for a more realistic and accuracy exposure assessment in the future, based on data gaps we identify.     

Inactivation effect of newly developed low concentration electrolyzed water and other sanitizers against microorganisms on spinach

The efficacy of newly developed low concentration electrolyzed water (LcEW) was investigated to inactivate the pathogens on spinach leaves as a convenient and safe alternative sanitizer and it was compared to other sanitizers. Spinach leaves were inoculated with Escherichia coli O157:H7 and Listeria monocytogenes and dip treated with deionized water (DIW), LcEW, strong acid electrolyzed water (SAEW), aqueous ozone (AO), 1% citric acid (CA) and sodium hypochlorite solution (NaOCl) for 3 min at room temperature (23 ± 2 °C). For all pathogens, the similar pattern of microbial reduction on spinach was apparent with LcEW and SAEW washing. In the present study, it was found that LcEW inactivated, at maximum, 1.64–2.80 log cfu/g and DIW resulted in lowest reduction, 0.31–0.95 log cfu/g of background or pathogenic microflora present on spinach leaves compared to the unwashed control. The findings of this study indicate that LcEW and SAEW did not differ significantly (P > 0.05) in reducing background or pathogenic microflora on spinach and LcEW may be a promising sanitizer for washing vegetables without environmental pollution instead of using electrolyzed oxidizing (EO) water or SAEW.     

Influence of the hydrophobicity and surface roughness of mangoes and tomatoes on the adhesion of Salmonella enterica serovar Typhimurium and evaluation of cleaning procedures using surfactin

Salmonellosis is associated with the consumption of raw vegetables and fruits such as tomatoes, watermelons, alfalfa sprouts, radishes, carrots, lettuce and parsley. The influence of the fruits' roughness on bacterial adhesion was evaluated as measured using a profilometer. The adhesion of Salmonella enterica serovar Typhimurium to mango and tomato surfaces was also evaluated by measuring of the hydrophobicity of the microorganisms and the fruits surfaces. The bacteria adherent on fruit's surface was quantified by plate count and visualize by scanning electron microscopy. In addition, the efficiency of surfactin in removing S. Typhimurium from the fruits' surfaces was analyzed. The average roughness (R_a) of mango (4.54 ± 1.95 μm) was significantly different (p < 0.05) compared to tomato (2.88 ± 2.15 μm). The adhesion of the microorganisms to the fruits' surfaces, as predicted by a determination of the total energy of adhesion (ΔG), was thermodynamically unfavorable. Despite these data, the numbers of bacteria on both fruits' surfaces were similar (p > 0.05), reaching 5.95 ± 0.36 log CFU cm⁻² and 5.81 ± 0.39 log CFU cm⁻² on mango and tomato, respectively. Therefore, these results suggest that the adhesion observed in this experiment is a multifactorial process. Surfactin removed 94.3% and 92.2% of the S. Typhimurium adhered to the surfaces of the mangoes and tomatoes, respectively. Our research showed that the roughness and hydrophobicity of the fruits' surface did not affect the efficiency of each sanitation treatments on removing of S. Typhimurium. It was observed that the chlorine was more efficient treatment (p < 0.05) for tomato surface. For surface of mangoes, chorine and surfactin were better than water treatment for bacteria control.     

Microorganisms associated with the spoilage of Pupuru

This study involved evaluation of physicochemical and microbiological changes in dried Pupuru (fermented, smoke dried cassava) balls under storage conditions simulating those currently used by the traditional processors. The aim was to understand the process of spoilage with a view to reducing the rate. pH ranged from 3 to 4, reducing significantly in cabinet-dried samples from 4.24 to 3.27 after 6 days of storage. Viable counts were in the range of 6–8 log cfu/g. Spoilage microorganisms included aerobic spore-forming and non-sporing bacteria as well as potentially toxigenic moulds like Aspergillus flavus and Penicillium species, which could constitute a health hazard to consumers.     

A survey on the microbiological and chemical composition of buffalo milk in China

One hundred and twelve samples of raw buffalo milk were collected at four locations in China, and their microbiological and chemical composition was analyzed. Average levels of major components were: fat 7.59% (v/v), crude protein 4.86% (w/w), lactose 4.74% (w/w), total solids 18.44% (w/w), ash 0.85% (w/w), and pH 6.65. The crude protein, fat, total solids and ash contents of milk from multi-crossbreed buffalo were higher than those of river buffalo’s, but lower than those of crossbreed F₁ and F₂ buffalo’s. Microbiological enumeration revealed counts of total mesophilic aerobic bacteria of 5.59 log cfu/ml, bacterial endospores 2.31, lactic acid bacteria 4.62, fungi 1.79, coliforms 2.42, Escherichia coli 1.53, and Staphylococcus aureus 1.68. Listeria spp. were below detection level in all of the samples. The microbiological quality of buffalo milk was judged marginal and indicates the need for improved hygienic standards.     

Antimicrobial activity of clove (Syzgium aromaticum) oil in inhibiting Listeria monocytogenes on chicken frankfurters

The ability of Listeria monocytogenes to survive and grow at refrigeration temperature in some ready to eat (RTE) poultry products is a public health concern. The inhibitory effect of clove oil (1% and 2%, v/w) applied to the surface of RTE chicken frankfurters was determined on seven strains of L. monocytogenes inoculated at low (10²–10³ cfu/g) or high cell numbers (10⁴–10⁶ cfu/g), and stored at 5 °C for 2 weeks or at 15 °C for 1 week. All strains of L. monocytogenes survived and grew on control frankfurters at 5 °C and 15 °C but growth was inhibited under both storage conditions in the presence of either 1% or 2% clove oil. Depending on the sensory considerations, the addition of clove oil to frankfurters may be an effective strategy to control L. monocytogenes in chicken frankfurters.     

Antimicrobial effect of koruk (unripe grape—Vitis vinifera) juice against Salmonella typhimurium on salad vegetables

To evaluate the antimicrobial activity of koruk (unripe grape—Vitis vinifera) juice, cucumber and parsley samples were inoculated with two Salmonella typhimurium strains namely S. typhimurium NRRL-B-4420 and S. typhimurium CCM 583 at the level of approximately 10⁶ cfu/ml. The samples were treated with koruk juices produced by different methods (by using mortar and electrical blender) for 0, 15, 30 and 60 min. Salmonella cells were counted by using direct surface plating (on Tryptone Soya Agar, Bismuth Sulphite Agar or Xylose Desoxylate Agar with 50 ppm nalidixic acid; TSAN, BSAN or XLDN) and overlay plating (on TSAN/BSAN) methods. Counts on TSAN/BSAN or TSAN, BSAN and XLDAN were not significantly different (P > 0.05). So, further experiments were done with direct surface plating on TSAN and BSAN. Overall statistical analysis indicated that, the antimicrobial effect of koruk juice was found to be dependent on the culture strains and products used (P < 0.05). Koruk juice exerted an immediate antimicrobial effect contact with the test microorganisms causing 1–1.5 log cfu/g reductions in initial populations. However, cell reductions in samples exposed to koruk juice for 15, 30 and 60 min varied between 2 and 3.5 log cfu/g depending on the microorganism and test sample used. There was no significant difference in cell reduction in samples exposed to koruk juices for 15, 30 and 60 min (P > 0.05) whereas reduction obtained at 0 time differed significantly (P < 0.05). On the other hand, no significant differences were found between the antimicrobial effects of koruk juices prepared by different methods. Results of the study showed that koruk juice used as flavouring and acidifying agent in Turkish kitchens, may be considered to be an alternative potential antimicrobial agent at household level for salad vegetables.     

Microbial shelf-life extension of chilled Coho salmon (Oncorhynchus kisutch) and abalone (Haliotis rufescens) by high hydrostatic pressure treatment

This study was carried out to evaluate the effect of high hydrostatic pressure (HHP) treatment on microbial behavior and the shelf-life extension of Coho salmon and abalone during chilled storage at 4 °C. For salmon, HHP treatments were applied at 135, 170, and 200 MPa for 30 s, while abalone treatment consisted of 500 MPa for 8 min and 550 MPa for 3 or 5 min. Spoilage bacteria (Pseudomonas spp. and hydrogen sulfide-producing bacteria, mainly Shewanella putrefaciens), as well as aerobic mesophilic and psycrophilic microorganisms, were enumerated immediately after treatment and throughout subsequent storage at 4 °C. Results have shown that HHP treatment reduced the initial microbial counts of salmon from 3.16 to 2.2 log units, moreover abalone was reduced from 1.3 log to undetectable levels (<10 CFU g⁻¹). HHP-treatment used for salmon were not sufficient to extend their shelf-life. However, the shelf-life of abalone was extended from 30 (control samples) to >65 days irrespective of HHP treatment applied.     

Effect of combined ozone and organic acid treatment for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mushroom

This study evaluated the effects of ozonated water (1, 3, and 5 ppm) alone with different exposure times (0.5, 1, 3, or 5 min), and combinations of 3 ppm ozone with 1% organic acids (acetic, citric, or lactic acids) during 5-min exposure for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mushroom and to observe the regrowth of these pathogenic bacteria on treated enoki mushroom during storage for 10 days at 15 °C. Results showed that ozone treatment gave less than 1.0- and 0.5-log count reductions on E. coli O157:H7 and L. monocytogenes, respectively. Efficacy was improved with combined 3 ppm ozone and 1% citric acid treatment, resulting in 2.26- and 1.32-log count reductions, respectively. During storage at 15 °C (10 days) after combined treatment and packaging, populations of E. coli O157:H7 and L. monocytogenes increased to more than 8.0 log cfu/g, indicating that the combined treatment did not have a residual antimicrobial effect during storage. Although the storage study did not show control of these pathogens, the combined ozone–organic acid treatment was more effective than individual treatments in reducing initial population levels of these pathogens on enoki mushroom.