Ocurrence of Vibrio spp., positive coagulase staphylococci and enteric bacteria in oysters (Crassostrea gigas) harvested in the south bay of Santa Catarina island, Brazil

The aim of this study was to assess the contamination of oysters (Crassostrea gigas), harvested in six different regions of the South Bay of Santa Catarina Island, with Coliforms at 45 ºC, Escherichia coli, Vibrio spp., positive coagulase staphylococci, and Salmonella sp. over a period of one year. One hundred eighty oyster samples were collected directly from their culture sites and analyzed. Each sample consisted of a pool of 12 oysters. All of the samples analyzed showed absence of Salmonella, 18 (10%) samples showed presence of Escherichia coli, 15 (8.3%) samples were positive for V. alginolyticus, and Vibriocholerae was detected in 4 samples (2.2%). The counts of positive-coagulase staphylococci varied from <10 to 1.9 x 102 CFU.g-1, whereas the counts of Coliforms at 45 ºC and E. coli ranged from <3 to 1.5 x 102 MPN.g-1 and <3 and 4.3 x 10 MPN.g-1, respectively. Counts of V. parahaemolyticus and V. vulnificus ranged between <3 and 7 MPN.g-1, for both microorganisms. This suggests the need for monitoring these Vibrios contamination in oysters. Based on the results of the microbiological assays, the samples analyzed showed acceptable bacteriological quality, i.e., they were within the parameters established by Brazilian Legislation.

Microbiota of sausages obtained by spontaneous fermentation produced in the South of Brazil

The study of the ecology of fermented sausage is fundamental to understand the physical and chemical changes that happen during fermentation and maturation. The aim of the present study was to determine the microbiological characteristics of sausages produced by spontaneous fermentation. Fifty samples of sausages produced in the South of Brazil by different small manufacturers were analyzed for the following microbiota: aerobic mesophilic bacteria; Micrococcaceae; mold and yeast; lactic acid bacteria; total and fecal coliforms; coagulase-positive Staphylococcus, and Salmonella. In most samples (72%), the count of lactic bacteria was higher than 6 log10 cfu.g-1, and the samples with the highest counts were above 8 log10 cfu.g-1. The counts of Micrococcaceae in most samples were between 5 log10 and 7 log10 cfu.g-1. With respect to the presence of molds and yeasts, there was a significant variation among the samples with counts ranging from 2 log10 cfu.g-1 and 6 log10 cfu.g-1. From the data obtained, it was possible to conclude that 24% of the analyzed samples did not comply with the current law in Brazil since the levels of fecal coliforms or coagulase-positive Staphylococcus exceeded the maximum limit allowed.

Physicochemical and microbiological evaluation of corrientes artisanal cheese during ripening

The aim of this study was to evaluate some physical and chemical parameters (total solids, pH, acidity, fat, acid degree value of fat, salt, protein and nitrogen fractions) and their effects on the beneficial (lactic acid bacteria: LAB) and undesirable microbial populations (coliforms, Escherichia coli, Staphylococcus aureus, moulds, and yeast) during ripening of Artisanal Corrientes Cheese, an Argentinian cow's milk variety, to determine whether a longer ripening period than usual improve its hygienic-sanitary quality. The protein content was much higher than that of other cow's milk cheeses with similar values of fat. The larger peptides showed values three times higher in the 30 day-old cheese than those obtained in the beginning of the process. Staphylococcus aureus and Escherichia coli were detected (3.04 ± 1.48 log10 cfu/g of cheese, 2.21 ± 0.84 log10 MPN/g of cheese) even at 15 and 30 days of ripening, respectively. The distribution of three hundred LAB strains classified to the genus level (lactococci:lactobacilli:leuconostocs) was maintained during the ripening period. The high number of LAB in rennet may have contributed to the fermentation as a natural whey starter, unknown source of LAB for this specific cheese so far. The physicochemical changes that occur during ripening were not big enough to inhibit the growth of undesirable microorganisms.

Acetic acid as an intervention strategy to decontaminate beef carcasses in mexican commercial slaughterhouse

Beef can be contaminated during the slaughter process, thus other methods, besides the traditional water washing, must be adopted to preserve meat safety. The objective of this study was to evaluate the effect of 2% acetic acid interventions on the reduction of indicator bacteria on beef carcasses at a commercial slaughterhouse in Mexico. Reduction was measured by the count of mesophilic aerobic bacteria (TPC), total coliform (TC), and fecal coliform (FC) (log CFU/ cm²). Among the different interventions tested, treatments combining acetic acid solution sprayed following carcass water washing had greater microbial reduction level. Acetic acid solution sprayed at low pressure and longer time (10-30 psi/ 60 s) reached higher TPC, TC, and FC reductions than that obtained under high pressure/ shorter time (1,700 psi/ 15 s; P<0.05). Exposure time significantly affected microbial reduction on carcasses. Acetic acid solution sprayed after carcass washing can be successfully used to control sources of indicator bacteria on beef carcasses under commercial conditions.

Characterization of spoilage bacteria in pork sausage by PCR-DGGE analysis

To investigate microbial diversity and identify spoilage bacteria in fresh pork sausages during storage, twelve industrial pork sausages of different trademarks were stored at 4 ºC for 0, 14, 28 and 42 days, 80% relative humidity and packaged in sterile plastic bags. Microbiological analysis was performed. The pH and water activity (a w) were measured. The culture-independent method performed was the Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE). The culture-dependent method showed that the populations of mesophilic bacteria and Lactic Acid Bacteria (LAB) increased linearly over storage time. At the end of the storage time, the average population of microorganisms was detected, in general, at the level of 5 log cfu g-1. A significant (P < 0.005) increase was observed in pH and a w values at the end of the storage time. The PCR-DGGE allowed a rapid identification of dominant communities present in sausages. PCR-DGGE discriminated 15 species and seven genera of bacteria that frequently constitute the microbiota in sausage products. The most frequent spoilage bacteria identified in the sausages were Lactobacillus sakei and Brochothrix thermosphacta. The identification of dominant communities present in fresh pork sausages can help in the choice of the most effective preservation method for extending the product shelf-life.

Survival of Lactobacillus rhamnosus probiotic strains in peach jam during storage at different temperatures

The survival of six probiotic wild strains of Lactobacillus rhamnosus was compared with that of a type strain during 78 days of storage at 25 and 5 ºC in peach synthetic medium (PSM) and commercial peach jam (PJ). Changes in viable cell counts, pH values, sugar content, and colour parameters were monitored. All strains exhibited better performances in PJ than in PSM, showing count values higher than 7 Log cfu g-1 up to 78 days of storage at 5 ºC. Almost all wild strains remained above the critical value of 6 Log cfu g-1 in samples stored at 25 ºC up to 45 days, while the Lb. rhamnosus GG type strain, used as control, was not able to survive later than 15 days. In the synthetic medium used, the strains showed better survival in the samples incubated at 25 ºC, remaining viable above the critical level up to 45 days of storage, except for the strain H12. The probiotic cultures added to jam did not significantly change the colour parameters of the product; however the metabolism of lactobacilli did cause changes in the pH and in the composition of sugars.

Salted lamb meat blanket of Petrolina-Pernambuco, Brazil: process and quality

Salted lamb meat blanket, originated from boning, salting, and drying of whole lamb carcass, was studied aiming at obtaining information that support the search for guarantees of origin for this typical regional product from the city of Petrolina-Pernambuco-Brazil. Data from three processing units were obtained, where it was observed the use of a traditional local technology that uses salting, an ancient preservation method; however, with a peculiar boning technique, resulting in a meat product with great potential for exploitation in the form of meat blanket. Based on the values of pH (6.22 ± 0.22), water activity (0.97 ± 0.02), and moisture (69.86 ± 2.26) lamb meat blanket is considered a perishable product, and consequently it requires the use of other preservation methods combined with salt, which along with the results of the microbiological analyses (absence of Salmonella sp, score <10 MPN/g of halophilic bacteria, total coliforms between 6.7 × 10³ and 5.2 × 10(6) FUC/g, and Staphylococcus from 8.1 × 10³ CFU/g at uncountable) reinforce the need of hygienic practices to ensure product safety. These results, together with the product notoriety and the organization of the sector are important factors in achieving Geographical Indication of the Salted lamb Meat blanket of Petrolina.

Frog meat microbiota (Lithobates catesbeianus) used in infant food

Captive breeding of bullfrog (Lithobates catesbeianus) is of great economic potential, mainly for its thighs and leather. The nutritional quality of frog meat includes properly balanced amino acids with a protein profile of high biological value, low fat and low cholesterol, and high digestibility due to its short chain molecule structure. It is recommended by doctors and nutritionists, especially for protein restricted children or malnourished children. Aiming to aggregate value to the segment and offer a product with nutritional properties that meet the need of children aged six months and above, a meat product based on the composition of frog meat was developed experimentally. To ensure raw material quality after bleaching and deboning, the microbiota present in the frog meat was determined. The analyses were performed according to Brazilian laws. It was observed that the resident and transient microbiota met the standards set by regulations. The results found were: mesophyll 4.5 x 10(4) CFU/g; Staphylococcus coagulase positive 2.0 x 10² CFU/g; negative for Salmonella sp. and Aeromonas spp. The findings indicate that the raw material showed satisfactory sanitation even in terms of family industry.

Quality of eggs sold in different commercial establishments and the study of the conditions of storage

The objective of this study was to investigate the influence of storage conditions on the physicochemical (mass, albumen height, pH, and Haugh unit) and microbiological (Salmonella spp., determination of the Total and Thermotolerant Coliforms, and Counting of the Mesophilic Aerobic Bacteria) quality of eggs. In the first experiment, a questionnaire was applied, and 33 samples of eggs were collected for the Salmonella spp analysis. In the second experiment, the eggs were collected from supermarkets, open markets, and distributors for physicochemical analysis. In the third experiment, 175 eggs were collected from the distributor, packaged in cardboard boxes lined with plastic wrapping, stored at 5 °C and 28 °C, and the physicochemical and microbiological analyses were performed at 7, 14, and 21 days of storage. In the first experiment, 100% of the samples analyzed showed no Salmonella spp. In the second experiment, it was found that the values of physicochemical parameters were in agreement with those in the literature. However, in the third experiment, the physicochemical parameter results showed statistical difference during storage and temperatures studied. Salmonella spp. were found in the samples stored at room temperature and in the refrigerated samples. Mesophilic microorganisms with values ranging from <10 CFU g- 1 (estimated) to 8.0 × 10³ CFU g- 1 and coliforms to 4 NMP.g- 1 were also found, but the presence of E. coli was not confirmed.

Production of probiotic fresh white cheese using co-culture with Streptococcus thermophilus

In this research, the probiotic Streptococcus thermophilus was inoculated into milk as co-culture to produce probiotic cheese. The effects of using Streptococcus thermophilus with other probiotic bacteria on cheese composition, and microbiological viability during 28 days of storage were investigated. Sensorial properties were determined only at 1st and 28th days of storage. The results showed that the use of Streptococcus thermophilus as co-culture in probiotic cheese production did not affect negatively the cheese components. Fat and dry matter properties of cheese weren't influenced by added probiotic bacteria. However, different level of pH, salt and lactic acid were detected. All probiotic bacteria were present in high levels throughout storage of cheeses, above 7 Log cfu.g- 1, threshold required for probiotic activity. Sensory panel showed that the highest average sensory evaluation points were recorded in cheeses made with Streptococcus thermophilus plus Lactobacillus casei, whereas other probiotic bacteria combinations had been affected less in regard to taste or appearance.

Ability of a Lactobacillus rhamnosus strain cultured in milk whey based medium to bind aflatoxin B1

This study aimed to compare Lactobacillus rhamnosus growth in MRS (de Man, Rogosa and Sharpe) broth and a culture medium containing milk whey (MMW) and to evaluate aflatoxin B1 (AFB1) adsorption capacity by bacterial cells produced in both culture media. L. rhamnosus cells were cultivated in MRS broth and MMW (37 °C, 24 hours), and bacterial cell concentration was determined spectrophotometrically at 600 nm. AFB1 (1 µg/ml) adsorption assays were conducted using 1 x 10(10) non-viable L. rhamnosus cells (121 °C, 15 minutes) at pHs 3.0 and 6.0 and contact time of 60 minutes. AFB1 quantification was performed by High Performance Liquid Chromatography. Bacterial cell concentration in MMW was higher (9.84 log CFU/ml) than that in MRS broth (9.63 log CFU/ml). There were no significant differences between AFB1 binding results at the same pH value (3.0 or 6.0) for the cells cultivated in MRS broth (46.0% and 35.8%, respectively) and in MMW (43.7% and 25.8%, respectively), showing that MMW can adequately replace the MRS broth. Therefore, it can be concluded that the use of L. rhamnosus cells cultivated in MMW offers advantages such as reduction in large scale production costs, improvement of environmental sustainability, and being a practicable alternative for decontamination of food products susceptible to aflatoxin contamination.

Molecular screening of bovine raw milk for the presence of Shiga toxin-producing Escherichia coli (STEC) on dairy farms

Milkborne transmission of Shiga toxin- producing Escherichia coli (STEC) has raised considerable concern due to recent outbreaks worldwide and poses a threat to public health. The aim of this study was to develop a sensitive and specific multiplex PCR assay to detect the presence of STEC in bovine raw milk. To identify E. coli (ATCC 25922) contamination, the gene uspA was used, and PCR sensitivity and specificity were accessed by testing diluted samples ranging from 2 to 2.0 × 10(6) CFU/mL. To detect STEC, the stx1 and stx2 genes were selected as targets. After reaction standardization, the multiplex assay was tested in raw milk collected from 101 cows on dairy farms. PCR assay for E. coli detection had a specificity of 100% and sensitivity of 79% (P<0.0001), with a lower detection limit of 2 CFU/mL. Multiplex PCR assay had 100% sensitivity for E. coli positive raw milk samples, and 31.1% were contaminated with STEC, 28.3% of stx2, and 1.9% of stx1. The multiplex PCR assay described in the present study can be employed to identify and screen E. coli harboring stx1 and stx2 genes in raw milk on dairy farms and in industries.

Characterization of goat milk and potentially symbiotic non-fat yogurt

Combining prebiotics and probiotic microorganisms improve quality in the formulation of foods. In this paper, the characteristics of goat milk and symbiotic yogurt were studied. Raw goat milk was analyzed and the skimming process was optimized. For the formulation of a potentially non-fat symbiotic yogurt made with skimmed goat milk, inulin, gelatin, sugar, and Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus casei subsp. rhamnoshus. Chemical characteristics, acceptability, and viability of lactic acid bacteria and probiotic culture were assessed. The protein and fat content of the raw milk was 2.90 and 3.56 g/100 mL, respectively. The optimum skimming process was obtained at 9,800 rpm and 4 °C for 15 minutes. The product formulated had a protein and fat content of 4.04 to 0.04 g/100 mL, good sensory properties, and acceptability of 95%. The lactic bacteria count was 9 × 10(7) CFU mL- 1, and probiotic culture count was higher than 1 × 10(6) CFU mL- 1, which guarantees their effect and capacity to survive in the digestive tract and spread in the intestine. The yogurt was stable during the 21 days of storage. Therefore, this study shows that goat milk yogurt is an adequate delivery vehicle of the probiotic culture L. casei and inulin.

Use of starter cultures isolated from native microbiota of artisanal sausage in the production of Italian sausage

The most promising microorganisms for use as starter cultures are those isolated from the native microbiota of traditional fermented products. The aim of this study was to evaluate the use of lactic acid bacteria selected from the native microbiota of sausages produced by spontaneous fermentation as starter cultures for the production of sausage. Strains of Lactobacillus plantarum 503 and 341 have the potential for use as starter cultures in the manufacture of Italian sausage type. The population of lactic acid bacteria in sausages was >8 log CFU.g-1 during fermentation, which caused the pH to decrease to <4.5. This decrease in pH and the water activity of < 0.90 of sausages ensures the safety and preservation of this product. Sausages produced with these lactic cultures fulfill the requirements for microbiological quality and composition of Italian sausage type. Our results suggest the possibility of using these starter cultures for the production of sausages with peculiar characteristics that contribute to the identity of the product.

Antimicrobial activity of amurca (olive oil lees) extract against selected foodborne pathogens

The antimicrobial activity of a methanolic extract of amurca (olive oil lees) was determined against both Gram-positive (L. monocytogenes and S. aureus) and Gram-negative (E. coli O157:H7 and S. enteritidis) foodborne pathogens at 10 °C or 37 °C using microdilution and disk diffusion methods, and its relative activity was compared to selected antibiotics. Minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of amurca extract ranged from 60 to 80 µl/ml at 37 °C after 24 h against all tested strains. At 10 °C, amurca was more inhibitory with MIC and MBC values of 40 and 60 µl/ml, respectively, after 7 d against tested strains. Amurca at 40 µl/ml reduced numbers of tested pathogens by 2.5 to 3.2 log10 CFU/ml at 10 °C after 7 d, but was not inhibitory at 37 °C after 24 h. Protein prepared from amurca was not antimicrobial. The relative antimicrobial activity (inhibition zone ratio) of 80 µl/ml amurca methanolic extract compared to chloramphenicol, erythromycin, gentamycin and tetracycline ranged from 0.36 to 1.0 against Gram-negative and from 0.45 to 2.0 against Gram-positive bacteria. In addition, amurca extract inhibited E. coli O157:H7 02-0628 and S. aureus 26127 which were resistant to tetracycline and chloramphenicol, respectively.