INMACULADA VALLEJO MARINE AND ENVIRONMENTAL SCIENCE FACULTY UNIVERSITY OF CADIZ (SPAIN)

Transcription

1 INMACULADA VALLEJO MARINE AND ENVIRONMENTAL SCIENCE FACULTY UNIVERSITY OF CADIZ (SPAIN) Image fromhttp://es.wikipedia.org/wiki/playa_de_la_victoria

2 RELEVANCE AND NECESSITY OF A MICROBIAL ANALYSIS OF THE SEA WATER Microorganisms present in the sea water-risks: Direct exposition by recreational activities Indirect exposition by consuming marine organisms/products The infection depends on: 1. Presence of a pathogenic microorganism in the sea 2. Concentration of the pathogenic microorganism in the sea water 3. Capability of the microorganism to survive in the sea water 4. The infective threshold of the pathogenic microorganism

3 Microorganisms are able to modify the suitability of the sea for a given use such as recreation, fish life and/or marine aquaculture BACTERIAL ANALYSIS NEEDED INDICATOR MICROORGANISMS REQUIREMENTS FOR INDICATOR MICROORGANISMS 1. Cell growth has to be readily detected, quick isolation and identification 2. They have to be practically innocuous for humans and animals 3. Their presence has to be quantitative- and qualytative-related to the presence of other pathogenic microorganisms

4 INDICATOR MICROORGANISMS COLIFORMS COLIFORM IS NOT A TAXONOMIC BUT A WORKING DEFFINITION GRAM NEGATIVE FACULTATIVE ANAEROBIC ROD-SHAPE BACTERIA GROWTH AND LACTOSE FERMENTATION AT 35 ºC WITHIN H TOTAL COLIFORMS FAECAL COLIFORMS ALL COLIFORMS SUBSET OF TC THAT CAN ALSO FERMENT LACOTOSE AT 44,5 ºC WITHIN 48 H Escherichia and Klebsiella

5 FAECAL STREPTOCOCCI GRAM POSITIVE COCCI CHAINS OF VARYING LENGTH HUMAN PATHOGEN SPECIES

6 CADIZ (SPAIN) Image from Jacques Descloitres, MODIS RRTeam, NASA/GSFC

7 BEACHES OF CADIZ Image from Copyright Ana Gonzalez y Carlos Parodi. Chiclana-2002

8 URBAN BEACHES Image from

9 MICROBIAL ANALYSIS OF THE SEA WATER OF THE BEACHES OF CADIZ ROYAL DECREE 734/88 OF 1 st OF JULY EUROPEAN UNION DIRECTIVE 76/160 (BATHING WATER DIRECTIVE) Area / Points of survey Sample collection conditions Growth methods Counting methods

10 La Victoria SURVEY Cortadura 2500 m 3900 m 40 m 60 m 3 survey points 3 survey points Santa María del Mar La Caleta 20 m 400 m 850 m 35 m 1 survey point 1 survey point Images from

11 SAMPLE COLLECTION Glass bottles of minimum 100 ml Analysis of non-cooling samples within 1 hour after collection Analysis of cooling samples within 30 hours after collection Laboratory Analysis Sheet

12 CONSEJERÍA DE SALUD Delegación provincial de Cádiz JUNTA DE ANDALUCIA PROVINCE: VILLAGE: CODE: HEALTH RISK CONTROL OF BEACHES AND COASTAL AREAS SHEET Nm. 1: SURVEY CONDITIONS DATE OF SURVEY: / /200_ SAMPLE: COOLING TIME SURVEYOR SIGNATURE POINT OF SURVEY NAME CODE SAMPLE IDENTIFICATION WEATHER 0. SUNNY 1. PARTLY CLOUDY 2. CLOUDY 3. RAIN WIND 0. NO WIND 1. NORTH, 2. SOUTH 3. EAST, 4.WEST WATER COLOUR OILS FOAMS SOLIDS TIDE HIGH LOW TARS SAND ALGAE SOLIDS OTHER OBSERVATIONS GENERAL OBSERVATIONS: GENERAL STATE OF THE BEACHES: WATER TRANSPARENCY: YES NO

13 GROWTH METHODS

14 SPECIFIC MEDIA AND INCUBATION CONDITIONS TOTAL COLIFORMS ENDO-Agar media (Schiff reactive) Temperature: 35 ºC Time: 24 hours METALLIC GREEN COLONIES

15 FAECAL COLIFORMS FC-Agar media (Anyline blue) Temperature: 44,5 ºC Time: 48 hours BLUE COLONIES

16 FAECAL COLIFORMS Enterococcus-Agar media Temperature: 35 ºC Time: 48 hours LIGHT RED COLONIES

17 COUNTING METHODS COUNTING OF NUMBER OF COLONIES COLONY Group of bacteria derived from the growth of a single bacteria or a single colony-forming unit (CFU) TOTAL BACTERIA COUNT Number of colonies or CFU, visible on 6-8x magnification, which develops from the bacteria present in 100 ml of water in plate cultures with nutrien media at affixed incubation temperature and within a defined period of time

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19 EUROPEAN UNION DIRECTIVE 76/160 (BATHING WATER DIRECTIVE) INDICATOR MICROBE GUIDELINE VALUES IMPERATIVE VALUES SURVEY FREQUENCY Total coliforms Bimonthly Faecal coliforms Bimonthly Faecal Enterococci When contamination is suspected

20 ANNUAL HEALTH REPORT ON WATER QUALITY OF THE BEACHES OF CADIZ BEACH SURVEY AREA WATER CLASSIFICATION ANALYTICAL RESULTS INCIDENCES LA VICTORIA DELFINES CRUZ ROJA Very god quality Very god quality According IV a for quality According IV for quality - - ISECOTEL Very god quality According IV for quality - EL CHATO Very god quality According IV for quality - CORTADURA CORTADURA Very god quality According IV for quality - RES. MILITAR Very god quality According IV for quality - STA. Mª DEL MAR STA. Mª DEL MAR Very god quality According IV for quality - LA CALETA LA CALETA Very god quality According IV for quality - a) IV: Imperative values

21 DIRECTIVE 2006/7/CE OF THE EUROPEAN UNION INDICATOR MICROBE EXCELLENT QUALITY GOOD QUALITY SUFFICIENT QUALITY METHODS FOR ANALYSIS Faecal Enterococci UFC/100 ml ISO or ISO Escherichia coli UDC/100 ml ISO or ISO

22 REFERENCES Atlas, R. and Bartha, R Ecología microbiana y microbiología ambiental. 4ª Ed. Addison Wesley, Madrid. Bacteriological Analytical Manual Online ( Blue Flag Organization ( Borrego, J. J Métodos microbiológicos rápidos para análisis de aguas y alimentos. Universidad de Málaga. Consejería de Salud de la Junta de Andalucía ( Directiva 76/160/CEE del Consejo, de 8 de diciembre de 1975, relativa a la calidad de las aguas de baño. Directiva 2006/7/CE del Parlamento Europeo y del Consejo, de 15 de febrero de 2006, relativa a gestión de la calidad de las aguas de baño y por la que se deroga la Directiva 76/169/CEE. Guide for analytical methods of the Provincial Laboratory of Cadiz Laboratorio provincial de Cádiz (Consejería de Salud de la Junta de Andalucía). Madigan, M. T., Martinko, J. M., and Parker, J Brock. Biología de los microorganismos. 10th Ed. Prentice Hall Iberia, Madrid. Prescott, L. M., Harley, J. P., and Klein, D. A Microbiología. McGraw- Hill Interamericana, Mexico D. F. Real Decreto 734/1988, de 1 de julio, por el que se establecen las normas de calidad de las aguas de baño.