Andréia Cirolini

Occurrence of potentially pathogenic Vibrio in oysters (Crassostrea gigas) and waters from bivalve mollusk cultivations in the South Bay of Santa Catarina

INTRODUCTION This research aimed to identify and quantify potentially pathogenic Vibrio from different cultivations of bivalve shellfish in the State of Santa Catarina, Brazil, and water regions in the South Bay, as well as correlate the incidence of these microorganisms with the physicochemical parameters of marine waters. METHODS Between October 2008 and March 2009, 60 oyster and seawater samples were collected from six regions of bivalve mollusk cultivation, and these samples were submitted for Vibrio counts. RESULTS Twenty-nine (48.3%) oyster samples were revealed to be contaminated with one or more Vibrio species. The Vibrio parahaemolyticus and Vibrio vulnificus counts in the samples ranged from < 0.5 log10 Most Probable Number (MPN) g(-1) to 2.3 log10 MPN g(-1) oyster and from < 0.5 log10 MPN g(-1) to 2.1 log10 MPN g(-1) oyster, respectively. Of the 60 seawater samples analyzed, 44 (73.3%) showed signs of contamination with one or more vibrio species. The counts of V. parahaemolyticus and V. vulnificus in the samples ranged from < 0.3 log10 MPN·100mL(-1) to 1.7 log10MPN·100mL(-1) seawater and from < 0.3 log10 MPN·100mL(-1) to 2.0 log10 MPN·100mL(-1) seawater, respectively. A positive correlation between V. vulnificus counts and the seawater temperature as well as a negative correlation between the V. parahaemolyticus counts and salinity were observed. CONCLUSIONS The results suggest the need to implement strategies to prevent vibrio diseases from being transmitted by the consumption of contaminated bivalve shellfish.

Depuration of Oysters (Crassostrea gigas) contaminated with Vibrio parahaemolyticus and Vibrio vulnificus with UV light and chlorinated seawater.

The efficacy of depuration using UV light and chlorinated seawater for decontaminating Vibrio parahaemolyticus and Vibrio vulnificus from oysters was investigated. Oysters were contaminated with a five-strain cocktail of V. parahaemolyticus or V. vulnificus to levels of 10(4) to 10(5) CFU ml(-1) for bioaccumulation. The depuration was conducted in a closed system in which 350 liters of seawater was recirculated at a rate of 7 liters/min for 48 h at room temperature. Counts of V. parahaemolyticus or V. vulnificus were determined at 0, 6, 18, 24, and 48 h. Three treatments were conducted: T1, control treatment; T2, UV treatment; and T3, UV plus chlorine treatment. After 48 h of depuration of V. parahaemolyticus, T3 reduced the count by 3.1 log most probable number (MPN) g(-1) and T2 reduced the count by 2.4 log MPN g(-1), while T1 reduced the count by only 2.0 log MPN g(-1). After 48 h of depuration of V. vulnificus, T2 and T3 were efficient, reducing the counts by 2.5 and 2.4 log MPN g(-1), respectively, while T1 reduced the count by only 1.4 log MPN g(-1). The UV light plus chlorine treatment was more efficient for controlling V. parahaemolyticus in oysters. Both UV light and UV light plus chlorine were efficient for V. vulnificus. The present study is the first report showing the efficacy of depuration systems for decontaminating V. parahaemolyticus and V. vulnificus in oysters cultivated on the Brazilian coast. This study provides information on processes that can contribute to controlling and preventing such microorganisms in oysters and could be used for effective postharvest treatment by restaurants and small producers of oysters on the coast of Brazil.

Métodos alternativos para contagem de micro-organismos em carcaças suínas

Micro-organisms are present throughout the meat processing chain, from raw materials until finished product. Industrial quality programs check for contamination in raw materials and final products to ensure the safety of consumer products; for this reason, the use of alternative methods instead of the conventional methods used in routine diagnostic laboratories is necessary to obtain reliable results in the shortest time possible. Samples of pig carcasses were collected in a slaughterhouse with Federal Inspection in the State of Santa Catarina, Brazil. Mesophilic micro-organisms, Enterobacteriaceae and Escherichia coli were evaluated and tests performed by alternative methods, TEMPO®, Petrifilm™, and the conventional method of counting micro-organisms in Petri dishes. After statistical analysis, the Pearson correlation coefficient (r) showed a strong correlation up to 0.70 between the methodologies used for the enumeration of Enterobacteriaceae and mesophilic micro-organisms, show a moderate correlation between 0.30 to 0.70, for the diagnosis of Escherichia coli. The use of alternative methods to replace the conventional method can be recommended for diagnosis of Escherichiacoli, Enterobacteriaceae and mesophilic micro-organisms, because there was consistency between the results and the speed was increased by the alternative methodologies benefiting the pork industry.

Evaluation of the PetrifilmTM and TEMPO® systems and the conventional method for counting microorganisms in pasteurized milk

New microbiological methods have been developed and commercialized, but their performance must be guaranteed. The aim of the present study was to evaluate the PetrifilmTM and TEMPO® systems compared to the conventional method for counting microorganisms in pasteurized milk. A total of 141 samples of pasteurized milk were analyzed by counting mesophilic aerobic, Coliforms at 35 oC, Coliforms at 45 oC, and Escherichia coli microorganisms. High correlation was found between the methods for counting Coliforms at 35 oC, but low correlation was found for counting mesophilic aerobic, Coliforms at 45 oC, and Escherichia coli. No significant statistical difference was found among the three methods for counting Coliforms at 35 oC; however, the mean counts of mesophilic aerobic, Coliforms at 45 oC, and Escherichia coli showed significant statistical difference. PetrifilmTM and TEMPO® systems had satisfactory results for Coliforms at 35 oC in pasteurized milk but low performance for mesophilic aerobic, Coliforms at 45 oC and Escherichia coli.

Evaluation of the PetrifilmTM EB and TEMPO® EB systems with ISO 21528-2: 2004 method for the count of Enterobacteriaceae in milk

The development of alternative microbiological techniques is driven by the necessity to meet the current needs to deliver rapid results in the manufacturing process of foods, but it is important that these methods be evaluated for each application. The objective of the present study was to assess the Petrifilm™ EB and the TEMPO® EB systems with ISO 21528-2:2004 for the count of Enterobacteriaceae in pasteurized and UHT milk samples. We analyzed the microflora of 141 pasteurized milk samples, 15 samples of artificially contaminated pasteurized milk and 15 samples of artificially contaminated UHT milk. Investigation of the method Petrifilm™ EB and ISO 21528:2 regression analysis showed a high correlation in the samples, r = 0.90 for the microflora of pasteurized milk, r = 0.98 for artificially contaminated pasteurized milk and r = 0.99 for the artificially contaminated UHT milk. In evaluating the system TEMPO EB ® method and ISO 21528:2 correlation was also significant in the analyzed samples, with r = 0.86 for the microflora of pasteurized milk, r = 0.96 for artificially contaminated pasteurized milk and r = 0.99 for artificially contaminated UHT milk. No statistically significant differences were observed between the three methods conducted to analyze artificially contaminated pasteurized and UHT milk at three inoculum levels. In conclusion, the Petrifilm™ EB system and the TEMPO® EB system may be an alternative to the ISO 21528-2:2004 for the Enterobacteriaceae assay for milk as because of the ease-of-operation and the time reduction achieved for conducting the microbiological assay using these systems.

Avaliação do sistema Petrifilm™ HS na contagem de coliformes em leite pasteurizado

Alternative microbiological methods present several advantages over the conventional methodologies, but their efficiency should be confirmed. The present study aimed at analyzing Petrifilm™ HS and Petrifilm™ EC systems, and comparing with the conventional methodology for counting coliforms at 35 oC in pasteurized milk. High correlations were found among these methodologies used for coliforms counting at 35 oC. Petrifilm™ HS system presented satisfactory results for counting coliforms at 35 oC in pasteurized milk.