Simbarashe Samapundo

Effect of water activity and temperature on growth and the relationship between fumonisin production and the radial growth of Fusarium verticillioides and Fusarium proliferatum on corn

The two major fumonisin-producing Fusarium species are Fusarium verticillioides and Fusarium proliferatum. The growth and fumonisin production of these two isolates on corn was studied at water activities (a(w)) between 0.860 and 0.975 and at temperatures between 15 and 30 degrees C. Growth rates (g, mm/day) were obtained by linear regression during the linear phase of growth. In general, growth rates for both isolates increased significantly (P < 0.05) with increases in a(w) and temperature. Both fumonisin production and radial growth (mycelial development) for both isolates increased with a(w) at all temperatures investigated, but the effect of temperature on this relationship was not obvious. The effect of temperature on fumonisin production at high a(w) values optimal for growth was only marginal, whereas at lower a(w) values the effect of temperature was more pronounced, with more fumonisin production occurring at temperatures not optimal for growth. The optimum temperature for fumonisin production was between 15 and 25 degrees C. For F. proliferatum, the optimum temperature for growth at all a(w) values, 30 degrees C, resulted in the poorest fumonisin production. For both isolates, the slowest initial rate of fumonisin production was at 15 degrees C, the temperature at which the slowest growth rates were obtained.

Incidence, diversity and toxin gene characteristics of Bacillus cereus group strains isolated from food products marketed in Belgium.

The major objectives of this study were to determine the incidence, diversity and characteristics of Bacillus cereus group spp. isolated from food products marketed in Belgium. The food products investigated in this study included cooked pasta, lasagna, béchamel sauce, bolognaise sauce, fresh minced beef, fresh-cut vegetables and raw basmati rice. B. cereus group spp. were detected in 56.3% (324 of 575) of the samples giving rise to 380 strains. The highest incidence (100%) occurred in the raw basmati rice. Although only 10 (2.6%) of the 380 isolates were determined to be psychrotolerant (able to grow at ≤7°C), 25 (6.2%), 189 (49.7%) and 334 (87.9%) isolates were able to grow at mild temperature abuse conditions of 8°C, 9°C and 10°C, respectively. The large diversity of the isolates obtained (overall and between isolates obtained from the same product type) was highlighted by the results of the (GTG)(5) PCR fingerprinting of 80 selected isolates. Sixty-one of these 80 isolates belonged to 15 distinct clusters (≥85% Pearson correlation) whereas the remaining 19 were each clustered separately. Further diversity was also found in the distribution of toxin genes as 16 different profiles were observed in the 80 selected isolates. Whilst none of 80 selected strains harboured the ces gene required for the production of the emetic toxin cereulide, 42 strains (52.5%) carried all seven genes required for the production of the diarrhoeal enterotoxins: haemolytic BL, non-haemolytic enterotoxin and cytotoxin K. The results of this study highlight not only the omnipresence but also the highly diverse ecology of B. cereus spp. within and across several food product types available on the retail market in Belgium. They should also provide the impetus for more studies to enable detailed risk assessment studies to be performed.

Effect of NaCl reduction and replacement on the growth of fungi important to the spoilage of bread.

The effect of NaCl and various NaCl replacers (CaCl(2), MgCl(2), KCl and MgSO(4)) on the growth of Penicillium roqueforti and Aspergillus niger was evaluated at 22 degrees C. In addition, challenge tests were performed on white bread to determine the consequences of NaCl reduction with or without partial replacement on the growth of P. roqueforti. From the results obtained it can be concluded that at equivalent water phase concentrations the isolates exhibited differing sensitivities to the salts evaluated with NaCl and MgCl(2) having the greatest inhibitory action on the growth of A. niger and P. roqueforti, respectively. MgSO(4) had the least antifungal activity. At equivalent molalities, CaCl(2) had in general the largest antifungal activity. Although the water activity (a(w)) lowering effects of the compounds studied play a large role in explaining the trends observed, at equivalent water phase concentrations MgCl(2) was found to have a smaller inhibitory effect on A. niger than that expected from its a(w) depressing effect. The challenge tests revealed that no difference occurred in the growth of P. roqueforti on standard white bread, bread with 30% less NaCl and bread in which 30% of the NaCl has been partially replaced by a mixture of KCl and Sub4Salt. These results are of importance in assessing the possible microbiological consequences of NaCl reduction or replacement in bread and similar bakery products.

Total mesophilic counts underestimate in many cases the contamination levels of psychrotrophic lactic acid bacteria (LAB) in chilled-stored food products at the end of their shelf-life.

The major objective of this study was to determine the role of psychrotrophic lactic acid bacteria (LAB) in spoilage-associated phenomena at the end of the shelf-life of 86 various packaged (air, vacuum, modified-atmosphere) chilled-stored retail food products. The current microbiological standards, which are largely based on the total viable mesophilic counts lack discriminatory capacity to detect psychrotrophic LAB. A comparison between the total viable counts on plates incubated at 30 °C (representing the mesophiles) and at 22 °C (indicating the psychrotrophs) for 86 food samples covering a wide range - ready-to-eat vegetable salads, fresh raw meat, cooked meat products and composite food - showed that a consistent underestimation of the microbial load occurs when the total aerobic mesophilic counts are used as a shelf-life parameter. In 38% of the samples, the psychrotrophic counts had significantly higher values (+0.5-3 log CFU/g) than the corresponding total aerobic mesophilic counts. A total of 154 lactic acid bacteria, which were unable to proliferate at 30 °C were isolated. In addition, a further 43 with a poor recovery at this temperature were also isolated. This study highlights the potential fallacy of the total aerobic mesophilic count as a reference shelf-life parameter for chilled food products as it can often underestimate the contamination levels at the end of the shelf-life.

Effect of high oxygen and high carbon dioxide atmosphere packaging on the microbial spoilage and shelf-life of fresh-cut honeydew melon.

This study evaluated the potential of modified atmospheres (MAs) combining high oxygen (O₂) and high carbon dioxide (CO₂) levels to extend the shelf-life of fresh-cut honeydew melon. Firstly, the effect of MA on the growth and volatile organic metabolite production of Candida sake, Leuconostoc mesenteroides and Leuconostoc gelidum, which had all been previously isolated from spoiled commercial fresh-cut honeydew melon, was evaluated separately on honeydew melon agar at 7 °C. Additionally, the effect of selected MAs on the microbial, physico-chemical and sensory quality of commercial fresh-cut honeydew melon cubes was evaluated at 7 °C. The results showed that MAs with high O₂ and high CO₂ levels greatly retarded the growth, CO₂ and volatile metabolite production (i.e. ethanol, 2-methyl-1-butanol, ethyl acetate, phenylacetic acid, nonanal) of C. sake on honeydew melon agar; especially MAs consisting of 50% O₂+50% CO₂ and 70% O₂+30% CO₂. In contrast, the MAs evaluated only had a minor effect on the growth and volatile metabolite production of L. mesenteroides and L. gelidum. Overall, the effect of MAs on colour, juice leakage, juiciness, and firmness of fresh-cut honeydew melon was small during storage. Sensory preference was generally for fresh-cut honeydew melon cubes packaged in MA of 50% O₂+50% CO₂. These were still acceptable on day five of storage and had appreciably lower populations of yeasts and lactic acid bacteria, lower quantities of volatile organic compounds, but slightly stronger colour oxidation compared to honeydew melon that was packaged in air. Additionally, most of the samples packed in air had blown by day five due to the large quantity of CO₂ production during storage. Therefore, 50% O₂+50% CO₂ is a potential MA solution for extending the shelf-life of fresh-cut honeydew melon.

Fumonisins exposure from freshly harvested and stored maize and its relationship with traditional agronomic practices in Rombo district, Tanzania

The exposure to fumonisins from the maize harvest of 2006 (before and after sorting and storage) for 67 families was assessed along with its statistical association with agronomic practices in Rombo, Tanzania. Fumonisins were determined by high-performance liquid chromatography (HPLC) and exposure estimated based on 60 kg body weight and maize intake of 356 g per person day−1. Based on contamination in the unsorted maize, exposure for 27% of the families would exceed the provisional maximum tolerable daily intake of 2 µg kg−1 body weight. The odds ratio of fumonisin exposure from maize hybrid 614 (H614) were: two times higher compared with non-H614 maize (p = 0.089); and five times higher when the maize was grown without fertilizer (p = 0.004). Sorting maize before storage reduced the percentage of families with unacceptable exposures to 6%. Therefore, the results imply that planting of the non-H614 seeds using fertilizers combined with pre-storage sorting of maize significantly minimizes fumonisins exposure in Rombo. A field experiment to verify these findings is needed.

Effect of O2CO2 enriched atmospheres on microbiological growth and volatile metabolite production in packaged cooked peeled gray shrimp (Crangon crangon)

This study evaluated the effect of modified atmospheres (MAs) with different O(2) concentrations on microbial growth and volatile metabolite production in gray shrimp (Crangon crangon) during storage at 4 °C. Eight MAs were evaluated in total. Four of the MAs evaluated were without CO(2): 0/0/100, 0/10/90, 0/30/70, 0/50/50 (% CO(2)/O(2)/N(2)) whilst the other four MAs all contained 50% CO(2): 50/0/50, 50/10/40, 50/30/20, 50/50/0 (% CO(2)/O(2)/N(2)). Volatile spoilage metabolites were identified by thermal desorption GC-MS and quantified during storage by selective ion flow tube mass spectrometry (SIFT-MS). In comparison to microbial growth observed with an atmosphere of 100% N(2), microbial growth was stimulated by the addition of O(2) in the MAP in the absence of CO(2.) Under these conditions the total psychrotrophic counts exceeded 7 log cfug(-1) after just 3 days of storage. However, in the presence of 50% CO(2) the total psychrotrophic count exceeded 7 log cfug(-1) after 5 days of storage. The combination of 50% CO(2) and 50% O(2) significantly inhibited microbial growth. For this MA condition, a diminishing effect on the production of metabolites was also observed, especially for amines and sulfur compounds, which constituted the major fraction of components causing the offensive odor.

The influence of headspace and dissolved oxygen level on growth and haemolytic BL enterotoxin production of a psychrotolerant Bacillus weihenstephanensis isolate on potato based ready-to-eat food products

The major objective of this study was to determine the influence of the initial headspace and dissolved O(2) level and vacuum packaging on growth and diarrhoeal enterotoxin production by Bacillus weihenstephanensis on potato based ready-to-eat food products. In general, the lower the initial headspace or dissolved O(2) level the slower the maximum growth rate (μ(max), log(10) CFU g(-1) d(-1)), the longer the lag phase duration (λ, d) and the smaller the maximum population density (N(max), log(10) CFU g(-1)) became. The slowest μ(max), the longest λ and the smallest N(max) were generally found for growth under vacuum packaging. This implies shorter shelf-lives will occur at higher initial headspace or dissolved O(2) levels as the growth of B. weihenstephanensis to the infective dose of 10(5) CFU g(-1) in such atmospheres takes a shorter time. Significant consumption of dissolved O(2) only occurred when growth shifted from the lag to the exponential phase and growth generally transitioned from the exponential to the stationary phase when the dissolved O(2) levels fell below ca. 75 ppb. Diarrhoeal enterotoxin production (determined via detection of the L2 component of haemolytic BL) was similar for growth under initial headspace O(2) levels of 1-20.9%, and was only reduced when growth took place under vacuum packaging. The reduction in L2 production when growth took place under vacuum was most probably related to the low final cell densities observed under this condition. Both growth and L2 production were inhibited over a 32-day incubation period at 7 °C by 40% CO(2) irrespective of the headspace or dissolved O(2) levels. The results illustrate the importance of residual O(2) and CO(2) on the shelf-stability and safety of modified atmosphere packaged potato based ready-to-eat food products with regards to B. weihenstephanensis.

Identification and characterization of yeasts causing chalk mould defects on par-baked bread

Pichia anomala, Hyphopichia burtonii and Saccharomycopsis fibuligera are spoilage yeasts causing chalk mould defects on par-baked breads packaged under modified atmosphere. The first objective of this study was to identify yeasts isolated from spoiled par-baked breads by means of a RAPD protocol and to determine the dominant spoilers amongst identified strains. The second objective was to determine the effects of water activity (a(w)) and pH value on the growth rates and lag phase durations of P. anomala, H. burtonii and S. fibuligera. 95% of the yeasts tested were identified as P. anomala and 5% as S. fibuligera, H. burtonii was not detected. In order to investigate the effect of a(w) and pH the growth of the three yeasts was tested within an a(w) range of 0.88-0.98 and a pH range of 2.8-8.0. P. anomala was able to grow from pH 2.8 to 8 without a clear optimum. S. fibuligera and H. burtonii showed a pH optimum for growth of 5. The optimum water activity for growth was different for the three strains and varied between 0.96 and 0.98. These growth data were further used to develop secondary models that describe the relationship between a(w) and the radial or colony growth rate (g, mm/d) or the lag phase duration (λ, d). The identification of the spoilage organisms and a good understanding of the effects of a(w) and pH on the growth behavior is essential for future development of adequate conservation strategies against chalk mould defects.

Influence of NaCl reduction and replacement on the growth of Lactobacillus sakei in broth, cooked ham and white sauce

The growth inhibiting effects of NaCl and selected simple salt replacers (CaCl(2), MgCl(2), KCl and MgSO(4)) on the growth of Lactobacillus sakei were studied in de Man Rogosa Sharpe broth at 7 degrees C over a water phase concentration of 0 to 6.4%. The divalent chloride salts (CaCl(2) in particular) generally had the largest antimicrobial activities at equivalent water phase concentrations, molalities or water activity (a(w)) values. MgSO(4) had not only the least antimicrobial activity but also the smallest a(w) depressing capacity. The results also showed that the antimicrobial effects of CaCl(2) were not fully accounted for by its a(w) depressing effects. Challenge tests performed on cooked ham and white sauce showed that reduction of NaCl levels by 28 and 33%, respectively, had no influence on the microbial stability of these products to L. sakei. Ultimately the study concluded that the microbiological consequences of the full or partial replacement of NaCl on the growth of L. sakei largely depend on the initial level of NaCl, the level of replacement and the nature of the salt replacer used. Altered stability to L. sakei is most likely given a high initial NaCl level, combined with a large level of partial replacement with either CaCl(2) (increased stability) or MgSO(4) (reduced stability) as the replacer.