Occurrence of storage fungi,especially aflatoxin-forming Aspergillus flavus, in soil,greenstuff and prepared hay

An investigation of the fungal flora in soil and greenstuff from a hayfield was carried out in order to show whether the occurrence of storage fungi, especially A. flavus, was sufficiently high to cause heavy infection of hay. Samples of the hay were field-dried to 63, 76 and 80% dry matter and stored in 150-kg containers in the barn. Three tons of the hay was field-dried to 60 and 70% dry matter and hayloft-dried. Field-dried and hayloft-dried hay were examined after 4 and $\text{4}\text{1}\text{2}$ months respectively. For comparison, hayrack-dried hay of an excellent quality of another origin was included and examined after 6 months of storage in a barn. Field-dried hay was heavily infected with Aspergillus and Penicillium, and in one case with Rhizopus. Hayloft-dried hay which was judged to be of good quality contained Aspergillus and Penicillium, but to much lesser extent. Hayloft-drying from 60% dry matter gave the best product. The hayrack-dried hay contained no A. flavus, only small amounts of Penicillium, and showed good agreement with the greenstuff fungal population. The greenstuff on the experimental field contained few A. flavus, with low aflatoxin-forming capability. From both field-dried and hayloft-dried hay, A. flavus with variable aflatoxin-forming capacity was frequently isolated.     

Survey of mycoflora counts, aflatoxin production and induced biochemical changes in walnut kernels

Fungal infection and mycotoxin contamination in fresh and stored kernels of walnut (Juglans regia) collected from different localities of Uttaranchal (India) were investigated. Fresh samples carry a combination of field as well as storage fungi. Species of Alternaria, Aspergillus and Penicillium were predominant. Thirty-nine percent of Aspergillus flavus isolates were toxigenic and produced up to 2170 μg/l of aflatoxin B₁ in the liquid media. Aflatoxin B₁ was the most common mycotoxin encountered as a natural contaminant in the stored samples. Twenty-one percent of fresh samples contained aflatoxin B₁ in low concentrations. The concentration of aflatoxin B₁ in fresh as well as stored samples was in the range of 140–1220 μg/kg. Characteristic rotting was observed in fresh as well as stored samples. The walnut kernels exhibited significant reductions in the levels of oil, starch and protein content during fungal infection.     

Population ecology of Aspergillus flavus associated with Mississippi Delta soils

Understanding the source of Aspergillus flavus is required to manage aflatoxin contamination of maize (Zea mays L.). Studies assessed A. flavus propagules, Fusarium spp., and total fungi associated with Mississippi Delta soils. Soils from 12 and 15 sites were collected in 2000 and 2001, respectively. The propagule density of A. flavus ranged from log(10) 2.0 to 4.3 colony-forming units (cfu) g^-1 soil, while total fusaria ranged from log(10) 3.0 to 5.4 cfu g^-1 soil. The highest populations of A. flavus were associated with soils containing higher organic matter, especially in sites under a no-tillage management. The frequency of aflatoxin production in isolates ranged from 13 to 81% depending on soil. In 2001, there was a highly significant correlation between A. flavus and the history of maize cultivation. Soil fertility factors such as organic matter content, nitrate and extractable phosphorus correlated with the density of Aspergillus, Fusarium spp., and total fungi. The relationship between soil parameters and Aspergillus populations may be useful in predicting the contribution of soil microflora to aflatoxin contamination.     

Spectral analyses of fluorescences in dent maize infected with Aspergillus flavus and Aspergillus parasiticus

Bright greenish yellow (BGYF) and blue white (BWF) fluorescences were associated with Aspergillus flavus and A. parasiticus infected maize. The fluorescences were studied spectrofluorometrically, the BGYF exhibiting a peak wave length between 480–485 nm and the BWF between 440–445 nm. Neither fluorescence varied in maize stored under different moistures and temperatures.

BWF was similar spectrally to the fluorescence of the endosperm of sound kernels but × 5 20 more intense. The spectrum of BWF was similar to Aflatoxin G₁ or a mixture of aflatoxin B₁, B₂, G₁ and G₂ when they were spotted on endosperm tissue. A color reference for BGYF was similar in peak wave length to BGYF. Amsoy soybeans without the seed coat fluoresced with a peak 470–475 nm and the intensity was low compared to BGYF in maize. A fluorescence of maize kernels visually similar to BGYF but not associated with Aspergillus infection or aflatoxin contamination was also investigated. This “false BGY” fluorescence was spectrally similar to the BGYF in infected kernels.     

Moulds and mycotoxins in sorghum stored in traditional containers in India

Various traditional containers have been used in India for storage of sorghum grains. Sorghum is susceptible to fungal infestation and toxin elaboration. The present study relates to the mould and mycotoxin contamination (aflatoxin B₁ and T-2 toxin) in stored sorghum in different storage containers viz. “Kotlu” (Storage rooms), earthenware pots, gunny bags and reed baskets. Aspergillus sp. and Fusarium sp. were the prominent genera and the “Kotlu” form of storage was most susceptible to fungal attack. Storage treatment had little effect on fungal contamination, but despite the fungal infestation, the mycotoxin contamination was found to be very low.     

Mycotoxin occurrence and Aspergillus flavus soil propagules in a corn and cotton glyphosate-resistant cropping systems

The effects of cotton-corn rotation and glyphosate use on levels of soil-borne Aspergillus flavus, aflatoxin and fumonisin contamination in corn and cotton seed were determined during 2002-2005 in Stoneville, Mississippi (USA). There were four rotation systems (continuous cotton, continuous corn, cotton-corn and corn-cotton) for both glyphosate-resistant (GR) and non-GR cultivars-herbicide system arranged in a randomized complete block design with four replications. Aspergillus flavus populations in surface (5-cm depth) soil, sampled before planting (March/April), mid-season (June) and after harvest (September), ranged from 1.47 to 2.99 log (10) cfu g^-1 soil in the four rotation systems. Propagules of A. flavus were higher in the continuous corn system compared to the continuous cotton system on three sample dates, and cotton rotated with corn decreased A. flavus propagules in three of nine sample dates. Propagules of A. flavus were significantly greater in plots with GR cultivars compared to non-GR cultivars in three samples. In cotton seed, aflatoxin and fumonisin levels were similar (≤4 µg kg^-1 and non-detectable, respectively) regardless of rotation and glyphosate. In corn grain, aflatoxin was above the regulatory level (≥20 µg kg^-1) only in GR cultivar in 2004 and 2005. Fumonisin was higher in non-GR cultivar (4 mg kg^-1) regardless of rotation in 2004; however, in 2002, 2003 and 2005, aflatoxin and fumonisin levels were similar regardless of rotation and glyphosate. These results indicate the potential for increased aflatoxin and fumonisin levels (1 of 4 years) in corn; however, climatic conditions encountered during this study did not allow for mycotoxin production. In laboratory incubation studies, fairly high concentrations of glyphosate were required to inhibit A. flavus growth; however no short-term effect of soil treatment with glyphosate on A. flavus populations were observed. These data suggest that altered populations of A. flavus or higher aflatoxin concentrations in corn grain were due to indirect effects of the GR cropping system.     

The effect of storage time and agroecological zone on mould incidence and aflatoxin contamination of maize from traders in Uganda

A study to determine mould incidence and aflatoxin contamination of maize kernels was carried out among dealers (traders) in the three agroecological zones of Uganda. The maize kernels were categorized into those stored for two to six months or for more than six months to one year. Results indicate that the mean moisture content of the kernels was within the recommended safe storage levels of ≤ 15% but was significantly lower in the Highland maize kernels followed by the Mid-Altitude (dry) kernels while the Mid-Altitude (moist) kernels had the highest levels. Across the agroecological zones, Aspergillus, Fusarium, Penicillium and Rhizopus were the most predominant fungal genera identified and, among their species, A. niger had the highest incidence, followed by A. flavus, F. verticillioides, A. wentii, A. penicillioides and Rhizopus stolonifer. There were more aflatoxin positive samples from the Mid-Altitude (moist) zone (88%) followed by those samples from the Mid-Altitude (dry) zone (78%) while samples from the Highland zone (69%) were least contaminated. Aflatoxin levels increased with storage time such that maize samples from the Mid-Altitude (dry and moist) stored for more than six months had mean levels greater than the 20 ppb FDA/WHO regulatory limits. Aflatoxin B₁ was the most predominant type and was found to contaminate maize kernels from all the three agroecological zones. These results indicate that maize consumers in Uganda are exposed to the danger of aflatoxin poisoning. Thus, there is the need for policy makers to establish and enforce maize quality standards and regulations related to moulds and aflatoxins across the agroecological zones to minimize health hazards related to consumption of contaminated kernels.     

The display life of retail-packaged beef steaks after their storage in master packs under various atmospheres

Beef strip loins were divided into four portions. One portion of each loin was vacuum-packaged and then stored at −1·5°C. The other portions were each divided into three steaks, which were retail-packaged. The retail packs were master-packaged under atmospheres of N₂, CO₂, or O₂ + CO₂ (2 : 1, v/v) and then stored at 2°C. Product was assessed after storage times of up to 60 days. At each assessment, a vacuum pack and a master pack of each type, each containing product from the same loin, were withdrawn from storage. The vacuum-packaged product was cut into three steaks, which were retail-packaged. The newly prepared retail packs and those from the master packs were displayed in a retail cabinet, at air temperatures that averaged between 3 and 5·7°C, and were assessed twice daily until the product was judged to be unacceptable. When first assessed, steaks cut from vacuum-packaged product were generally considered desirable, with little metmyoglobin in the surface pigment, although the edges of same steaks were discoloured. Steaks stored under N₂ or CO₂ for 4 days or less were only slightly desirable at best, with metmyoglobin forming relatively large fractions of the surface pigment. However, after storage under N₂ or CO₂ for 6 days or more, metmyoglobin fractions were low, and the steaks bloomed to a desirable red colour. Steaks stored under O₂ + CO₂ had lower metmyoglobin fractions, and were desirable after storage for up to 8 days. However, the fractions of metmyoglobin increased, and steaks were judged to be less desirable after longer storage times. Steaks stored under O₂ + CO₂ for 20 days were unacceptable. After storage, the numbers of bacteria on steaks from vacuum packs and N₂, CO₂, and O₂ + CO₂ atmospheres were, respectively, <10⁴, <10⁶, <10⁵, and <10⁴ CFU/cm². The flora from steaks stored under CO₂ were composed wholly of lactic acid bacteria. Other flora were dominated by lactic acid bacteria, but contained fractions of enterobacteria and/or Brochothrix thermosphacta.

The appearance of product from vacuum packs generally was unacceptable after 72 h of display. The display life of steaks stored under N₂ or CO₂ was shorter than that of the product from vacuum packs when product was stored for 2 days or less, or 46 days or more. After other storage times, the product from vacuum packs or master packs with N₂ or CO₂ atmospheres had a similar display life. The display life of product stored under O₂ + CO₂ was similar to that of product from vacuum packs or CO₂ or O₂ + CO₂ was similar to that of product from vacuum packs after storage times of 8 days or less but was shorter after storage times of 12 or 16 days. The flora on displayed product from vacuum packs or CO₂ or O₂ + CO₂. atmospheres did not attain the maximum number of 10⁷ CFU/cm². and the product did not develop off-odours of microbial origin. However, numbers of 10⁷ CFU/cm² were approached or attained during display of product stored under N₂ for 28 days or longer, and some of that product developed moderate off-odours. It then appears that, under temperature regimes that are common in commercial practice, retail-packaged strip-loin steaks with a display life of 2 days or longer can be obtained from master packs after storage periods of up to about 2, 4, or 7 weeks, respectively, with master-pack atmospheres of O₂ + COPin2 (2 : 1, v/v), N₂, or CO₂.     

The effect of glucose supplementation on the spoilage microflora and chemical composition of minced beef stored aerobically or under a modified atmosphere at 4 °C

Glucose was added to minced meat (pH 6.0) and stored under aerobic or a modified atmosphere (MA) composed of 80% O₂ and 20% CO₂ to assess the effects of carbohydrate on the microbial association and the chemical properties of the meat. The type of packaging affected the size and the final composition of the microflora. The microbial composition of the mince without added glucose in air or MA, given in log₁₀cfu/g respectively, were pseudomonads (9.8 and 7.3), Brochothrix thermosphacta (8.5 and 8.1), lactic acid bacteria (8.8 and 8.7), Enterobacteriaceae (7.2 and 6.1) and yeasts (4.3 and 4.2). In mince supplemented with 0.2% (w/w) glucose, similar composition and numbers were observed. Glucose, glucose 6-phosphate and lactic acid were consumed at slower rates by the flora on meat stored under MA than by the flora on meat stored in air. The addition of glucose enhanced gluconate production by the flora on meat stored in air. D-lactic and acetic acid were produced in all samples stored under the MA.     

Effect of gas barrier characteristics of films on aflatoxin production by Aspergillus flavus in peanuts packaged under modified atmosphere packaging (MAP) conditions

The effects of the gas barrier characteristics of three films (ASI, ASII and ASIII) and storage temperature on the growth of, and aflatoxin production by, Aspergillus flavus in peanuts packaged in air and under a modified gas atmosphere of CO₂:N₂ (65:35) were investigated. Mold growth was barely visible in air packaged peanuts using high-medium barrier films (ASI and ASII) and stored at 20°C with more extensive growth occurring in air packaged peanuts stored at 25 and 30°C. Extensive mycelial growth and sporulation occurred in all air packaged peanuts in a low barrier film (ASIII), especially at 30°C. Gas packaging inhibited mold growth in peanuts packaged in a high gas barrier film at 20°C. However, mold growth occurred in gas packaged peanuts packaged in film ASII at higher storage temperatures while extensive mycelial growth was observed in all peanuts packaged in film ASIII irrespective of storage temperature. Levels of aflatoxin greater than the regulatory limit of 20 ng/g were detected in all air packaged peanuts with the highest level of aflatoxin (76 ng/g) being detected in peanuts packaged in a high gas barrier film ASI. Aflatoxin production was inhibited in gas packaged peanuts using a high barrier film. However, higher levels of aflatoxin were detected in all gas packaged peanuts in medium-low gas barrier films (ASII and ASIII), particularly at higher storage temperatures. This study has shown that MAP using a CO₂:N₂ (65:35) gas mixture was effective in controlling aflatoxin production by A. flavus in peanuts to levels less than the regulatory limit of 20 ng/g. However, the antimycotic effect of low O₂-high CO₂ atmospheres is dependent on the gas barrier characteristics of the packaging films, especially at higher storage temperatures.     

Aspergillus flavus link and its occurrence in relation to other mycoflora on stored spices

Spices are used sparingly to flavour foods; their mycoflora and its possible role in food infection has not been thoroughly investigated. In an examination of 6 spices, Aspergillus flavus, other Aspergillus species, and species of Rhizopus, Mucor, Penicillium, Tricothecium, and Fusarium were found on seed surfaces. A. flavus which is implicated in the production of carcinogenic aflatoxin, was mainly found on ginger, mustard, garlic, and pepper. The highest fungal counts (10.6 × 10⁴/g) occurred in stored pepper (Piper nigrum L.) and the lowest in curry leaves (Murraya koenigii L.). Contamination of pepper and mustard with A. flavus seems to be related to the processing and storage of these spices.     

The effect of moisture level on high-moisture maize (Zea mays L.) under hermetic storage conditions—in vitro studies

Under humid and warm conditions harvested grains are susceptible to molding and rapid deterioration. Therefore, they should be dried to safe moisture levels that inhibit the activity of microorganisms. Drying to these moisture levels is not economical for farmers in developing countries. Preservation of grains at intermediate moisture levels under hermetic storage conditions could be feasible and economical in warm and moist climates.

The purpose of the current study was to examine the effect of various moisture contents (m.c.) on the quality of maize grains in self-regulated modified atmospheres during hermetic storage.

Maize at 14, 16, 18, 20 and 22% m.c. was initially conditioned for 28 days in tightly wrapped plastic bags and then stored in sealed containers at 30 °C for up to 75 days. Carbon dioxide produced within the containers replaced the oxygen. As the m.c. increased the time for O₂ depletion shortened, from 600 h at 14% m.c. to 12 h at 22%. The maize at 20 and 22% m.c. exhibited the highest dry matter (DM) losses, the lowest germination rates and the highest yeast and bacteria counts. The major fermentation product in the hermetically sealed maize was ethanol (0–5 g kg⁻¹ DM), along with lower concentrations of acetic acid (0–1 g kg⁻¹ DM).

The results obtained from the in vitro experiments indicate that maize at the tested moisture levels can be stored satisfactorily under sealed conditions in which self-regulated atmospheres provide protection against microflora damage. Further large-scale trials will be needed to evaluate the economic feasibility of storing high-moisture maize.     

Effect of freezing and storage on quality factors in Hamburg and leafy parsley

Two types of parsley — the Hamburg cv Berli ska and leafy type cv Paramount — were frozen and stored at temperatures of −20 and −30 °C for 9 months. One half of the material was blanched before freezing and the other half was non-blanched. In 100 g fresh leaves of Hamburg parsley there were 20.0 g of dry matter, 310mg of vitamin C, 7.5mg of β-carotene, 203mg of chlorophyll, 30.8 mg N---NO₃ and 0.078 mg N---NO₂. For the leafy type the corresponding values were 17.3 g, 257 mg, 9.4mg, 68.5mg, and 0.077mg. The material blanched before freezing showed significant losses in the contents of vitamin C (47–51%), nitrates (22–33%), and nitrites (43–55%) and distinctly smaller ones but also significant in the case of dry matter. During freezing and storage of frozen products there were losses in vitamin C, β-carotene, and chlorophyll while the levels of nitrates and nitrites were variable. Particularly great losses of vitamin C and β-carotene were observed in the non-blanched frozen leaves stored at −20 °C. After 9 months' storage, frozen products preserved 10–44% of vitamin C, 37–91% of β-carotene, 78–95% of chlorophyll, and 78–153% of nitrates. Of the types of parsley analyzed the Hamburg type was a better raw material for freezing because of a significantly higher content of vitamin C and chlorophyll and significantly less nitrates in frozen products. When the storage temperature was −30 °C, the blanching of leaves was not necessary, although it helped their pressing into cubes.     

Aflatoxin formation and gene expression in response to carbon source media shift in Aspergillus parasiticus

Aflatoxins are toxic and carcinogenic polyketide metabolites produced by fungal species, including Aspergillus flavus and A. parasiticus. The biosynthesis of aflatoxins is modulated by many environmental factors, including the availability of a carbon source. The gene expression profile of A. parasiticus was evaluated during a shift from a medium with low concentration of simple sugars, yeast extract (YE), to a similar medium with sucrose, yeast extract sucrose (YES). Gene expression and aflatoxins (B₁, B₂, G₁, and G₂) were quantified from fungal mycelia harvested pre- and post-shifting. When compared with YE media, YES caused temporary reduction of the aflatoxin levels detected at 3-h post-shifting and they remained low well past 12 h post-shift. Aflatoxin levels did not exceed the levels in YE until 24 h post-shift, at which time point a tenfold increase was observed over YE. Microarray analysis comparing the RNA samples from the 48-h YE culture to the YES samples identified a total of 2120 genes that were expressed across all experiments, including most of the aflatoxin biosynthesis genes. One-way analysis of variance (ANOVA) identified 56 genes that were expressed with significant variation across all time points. Three genes responsible for converting norsolorinic acid to averantin were identified among these significantly expressed genes. The potential involvement of these genes in the regulation of aflatoxin biosynthesis is discussed.     

The effects of residual oxygen concentration and temperature on the degradation of the colour of beef packaged under oxygen-depleted atmospheres

Samples of beef longissimus dorsi (LD), approximately 5 × 5 × 1 cm, were packaged in pairs under 10 litre volumes of N₂ or CO₂ containing O₂ at concentrations between 100 and 1000 ppm. The packaged samples were stored at temperatures of 5, 1, 0 or −1·5°C, for times between 4 and 48 h. Samples of beef psoas major (PM) were packaged under N₂ or CO₂ containing O₂ at between 100 and 600 ppm, and stored at −1·5°C for 24 or 48 h. After storage, each sample was assessed for colour deterioration and discoloration, and for the fraction of metmyoglobin in the surface pigment.

The results obtained with N₂ and CO₂ atmospheres were similar. The colours of all LD samples had deteriorated after 4 h storage at 5 or 1°C, although the degree of deterioration increased with increasing O₂ concentration. All LD samples stored for 12 h at 5 or 1°C were extensively discoloured, with metmyoglobin fractions generally exceeding 60%, but those stored at −1·5°C for 48 h or less, under O₂ concentrations ≤ 400 ppm had undergraded colours. The colours of some LD samples stored at −1·5°C under about 600 ppm of O₂ were also undergraded, but the colours of samples stored under 800 or 1000 ppm had deteriorated by 24 h. The colours of LD samples stored at 0°C under > 200 ppm had deteriorated after 24 h storage, and the colours of samples stored under 100 ppm O₂ had deteriorated after 48 h storage. All PM samples were wholly discoloured after storage at −1·5°C. Evidently, the colour of beef muscle of high colour stability is resistant to degradation by atmospheres containing < 600 ppm of O₂ when the meat is stored at sub-zero temperatures, but not when the storage temperature is at or above 0°C. Beef muscle of low colour stability, such as the PM, will discolour at all low concentrations of O₂ irrespective of the storage temperature.     

Effects of Fusarium toxin-contaminated wheat and feed intake level on the biotransformation and carry-over of deoxynivalenol in dairy cows

An experiment was carried out to examine the effects of feeding Fusarium toxin-contaminated wheat (8.21 mg deoxynivalenol (DON) and 0.09 mg zearalenone (ZON) per kg dry matter) at different feed intake levels on the biotransformation and carry-over of DON in dairy cows. For this purpose, 14 ruminal and duodenal fistulated dairy cows were fed a diet containing 60% concentrate with a wheat portion of 55% (Fusarium toxin-contaminated wheat (mycotoxin period) or control wheat (control period)) and the ration was completed with maize- and grass silage (50 : 50) on a dry matter basis. Daily DON intakes ranged from 16.6 to 75.6 mg in the mycotoxin period at dry matter intakes of 5.6-20.5 kg. DON was almost completely biotransformed to de-epoxy DON (94-99%) independent of the DON/feed intake, and the flow of DON and de-epoxy DON at the duodenum related to DON intake ranged from 12 to 77% when the Fusarium toxin-contaminated wheat was fed. In the serum samples, de-epoxy DON was detected in the range of 4-28 ng ml^-1 in the mycotoxin period, while concentrations of DON were all below the detection limit. The daily excretion of DON and de-epoxy DON in the milk of cows fed the contaminated wheat varied between 1 and 10 µg and between 14 and 104 µg, respectively. The total carry-over rates as the ratio between the daily excretion of DON and de-epoxy DON into milk and DON intake were in the ranges of 0.0001-0.0002 and 0.0004-0.0024, respectively. Total carry-over rates of DON as DON and de-epoxy DON into the milk increased significantly with increasing milk yield. In the urine samples, de-epoxy DON was the predominant substance as compared with DON with a portion of the total DON plus de-epoxy DON concentration to 96% when the Fusarium toxin-contaminated wheat was fed, whereas the total residues of DON plus de-epoxy DON in faeces ranged between 2 and 18% of DON intake in the mycotoxin period. The degree of glucuronidation of de-epoxy DON was found to be approximately 100% in serum. From 33 to 80% of DON and from 73 to 92% of de-epoxy DON, and from 21 to 92% of DON and from 86 to 100% of de-epoxy DON were glucuronidated in the milk and urine, respectively. It is concluded that DON is very rapidly biotransformed to de-epoxy DON in the rumen and only negligible amounts of DON and de-epoxy DON were transmitted into the milk within the range of 5.6-20.5 kg day^-1 dry matter intake and milk yields (fat corrected milk) between 10 and 42 kg day^-1.     

Microbiological effects of acid decontamination of pork carcasses at various locations in processing

The microbiological effect of hot (55° C), 1% (v/v) lactic acid sprayed on the surface of pork carcasses (n = 36) immediately after dehairing, after evisceration (immediately before chilling) or at both locations in slaughter/ processing was determined. Mean aerobic plate counts (APCs) of all acid-treated carcass surfaces were numerically lower than those of control carcasses: however, in most cases these reductions were not statistically significant (P>0·05). All samples tested for the presence of Salmonella and Listeria were negative. No significant differences in sensory characteristics or microbiological counts were evident for acid-treated and control carcass loins that were vacuum packaged and stored 0–14 days post-fabrication. Mean pH value and scores of sensory attributes such as lean color, surface discoloration, fat color, overall appearance and off-odor of chops from acid-treated carcasses were not significantly and/or consistently different from chops of comparable control carcasses. The role of bacterial attachment to pork skin and its effect on the decontaminating efficiency of lactic acid are discussed.     

Clostridium perfringens and its toxins in minced meat from Kars, Turkey

The aim of this study was to determine the prevalence of Clostridium perfringens and its toxins in minced meat. A total of 96 minced meat samples were collected from local markets (16) and small butcher's shops (80) in Kars (Turkey). Samples were analysed for the presence of C. perfringens and its toxins using a commercially available ELISA kit. A total of 31 (32%) Clostridium spp. strains were isolated and 17 (55%) of these isolates were identified as C. perfringens. Four (25%) of the samples from local markets and 27 (34%) from small butcher's shops were contaminated with Clostridium spp. Furthermore, C. perfringens was isolated from two (12%) and 15 (19%) samples from local markets and small butcher's shops, respectively. Mean counts of Clostridium spp. were 2.2 ± 0.83 × 10² CFU g^-1 for local markets and 4.35 ± 8.53 × 10² CFU g^-1 for small butcher's shops; mean counts for C. porringers were 2.75 ± 0.21 × 10² and 6.82 ± 10.96 × 10² CFU g^-1 from markets and butcher's shops, respectively. The number of samples contaminated with both Clostridium spp. and C. perfringens was higher in small butcher's shops than in local markets. Moreover, higher numbers of Clostridium spp. and C. perfringens were isolated in samples from small butcher's shops than from local markets. A total of 13 (13%) samples were also positive for toxins produced by the organism, as detected by ELISA. Eleven samples from small butcher's shops and two samples from local markets were positive for the C. perfringens toxins tested. Moreover, two (12%), one (1%), four (4%) and two (2%) samples were contaminated with C. perfringens types A, B, C and D, respectively. In conclusion, some meat samples collected from local markets and small butcher's shops contained C. perfringens and its toxins and, therefore, present a potential risk of food poisoning.     

Softness in dry-cured porcine biceps femoris muscles in relation to meat quality characteristics and processing conditions

The aim of the study was to quantify the effect of meat quality characteristics and some processing conditions on the softness of dry-cured biceps femoris (BF) muscles. The BF muscles were dissected from forty hams and classified according to their pH_BF into three groups: LpH (pH < 5.66), MpH (5.66  pH  6.00) and HpH (pH > 6.00). BF muscles within each pH_BF group were distributed into three different Salting levels (1%, 2% or 4% of added NaCl). Muscles were salted, vacuum-packed and stored at 3 °C for 30 days. The post-salting BF muscles were classified into two intramuscular fat (IMF) levels: Low (IMF < 4%) and High (IMF  4%). Thereafter, the muscles were divided into two pieces and dried at two of the three different Drying levels (1.5, 2 and 2.5 g H₂O/g desalted dry matter). Then, each piece was divided into two samples that were packed in N₂ and stored at 5 °C or 30 °C for 1 month. Stress Relaxation was used to evaluate texture. Dry-cured BF muscles with initial pH > 6.0, with IMF > 4% or with added NaCl levels less than 2% were more prone to show soft texture. Softness in dry-cured muscles can be reduced by applying an ageing temperature of 30 °C for 30 days, despite increasing proteolysis. The softness reduction by ageing at 30 °C compared with 5 °C is expected to be higher when applied to drier samples, which show a smaller increase in proteolysis.     

The storage life of chilled pork packaged under carbon dioxide

Pork cuts of longissimus dorsi muscle with overlaying fat and skin were packed under vacuum in film of low oxygen transmission rate, or under CO₂ in gas impermeable aluminium foil laminate. Cuts were stored at +3 or −1·5°C. Vacuum packaged cuts were grossly spoiled by Brochothrix thermosphacta after 2 weeks' storage at 3°C and after 5 weeks at −1·5°C. Cuts packaged under CO₂ were grossly spoiled by B. thermosphacta after 5·5 weeks' storage at 3°C. Growth of B. thermosphacta was suppressed when CO₂ packaged cuts were stored at −1·5°C. At that temperature, slow growth of enterobacteria was detected after a lag of about 18 weeks. The enterobacteria caused gross spoilage of an increasing proportion of cuts between 18 and 26 weeks. Muscle tissue with pale, soft, exudative (PSE) characteristics tended to lose colour after long storage periods, apparently because of loss of myogglobin with exudate. Until spoilage, the eating qualities of pork appeared little affected by prolonged storage.