"| WATERBORNE. PATHOGENS OP THE U.S. VIRGIN TSDANDS ae Project No. 02 a a oe it “ 20. ; 42-08-0001- 6-935 2 september 1906 -Pechnical Report ‘No. 25° “Caribbean . Research. Institute College of the Virgin Islands - ‘st. Thomas + U.S. Virgin: Islands - 00802 . 7 “submitted in ‘Partial ‘Pulfiliment . of Contract Obligations. tor wre Water Resources: Research Center. : “TECHNICAL ‘REPORT ‘NO. 25. -CARIBBEAN RESEARCH | INSTITUTE oo, COLLEGE OF THE VIRGIN ISLANDS ST. ‘THOMAS i U. Ss (VIRGIN. ISLANDS _ BOB02 PROJECT NO. 02 _ AGREEMENT NO. 14~08-0001-G-939 athe. “ peséatch’ on whitch - the report is: based” was financea in > | part. by. the. ‘United States Department. of. the Interior, Geological ‘Survey, “through the Virgin Islands “Water : we _. Resources Research Institute." "contents of this. publication do not” necessarily ‘reflect the -views and policies. of the: United States ‘Department . of 7 the Interior,. nor: does mention of trade: names. or ‘commercial products : constitute _ their) “endorsement. by the OU, See Government . eee oe gg Thee ABSTRACT AL séries of water quality analyses were carried out - during 1985 on drinking water sources’ in the U.S. Virgin “Islands. The object of the study was to determine — which _ human pathogens might be present, the implications. of their "presence, and the ‘value of current Environmental Protection “Agency indicators for predicting the presence of pathogens. | on water samples _ were taken from seven cisterns, four wells, and five ‘places along the public water. ‘line (16 total). A four ‘liter sample was | taken at. each site and- “returned "immediately to ‘the lab’ for processing. “Each | “sample. was” checked ‘for total and: fecal. coliform. using both multiple tube and membrane filter methods: approved by the American Public Health Association and U. S. Environmental - “Protection Agency. They were “cultured, where ‘applicable, standard enrichment | “broth. and on selective media. ae Individual colonies were removed and cultured on eight "differentiating media as well as on. the APL strips (Analytab Products, Plainview, N. Yes 20 tests). me Controls 7 weve - run simultaneously. using: ‘Blanks, (no. inoculum) and “pare: cultures of eight species from the ‘American Type es ‘culture Collection | (arc) . The results of these 1800 separate tests were collated and compared for consistency, occurrence of pathogens, and "reliability, of prediction from the coliform tests. Thirty "found. 1. “species: of. bacteria, ‘twenty Of: them: ‘pathogens, This, analysis showed that: All of. the 16 “sources chad human pathogenic a bacteria present. Fifteen ‘of the 16. sources also had fecal streptococcus. Present while = of the. contaminated sources. would pass. “current “coliform 1 EPA water. ; quality. standards. Use of a "pre- ~enrichment" technique is vital. to get a true estimate of the less vigorous . pathogens and injured ‘coliforms. | Current EPA indicators used for contami- nated afinking or groundwater are, at least in: ‘the. v. Tey useless for ‘predicting. the presence or. absence. of ‘pathogens. were Further comparison (of these results with earlier work “on ground and cistern water indicates ‘that the problem may ‘be more severe and ubiquitous than. previously thought. "Recommendations are ‘made for future management, de~ tection studies, and. control. CUES “ACK NOWLEDGEMENTS ae We appreciate. the guidance of. Dre ‘verry c. : “Hazen and, the assistance of the ‘University of Puerto Rico, Department of Biology, with ‘the. analysis of ‘Legionella spp. and for. "reading the manuscript; Dr. James — MeFetters | of ‘the _— University. of Montana and ‘Dre William Fox. of the v. Se EPA “for their interest and advice. we ‘thank the Department of - seience ’ and math, and the: Vv. re Cooperative. “Extension Service, college. of the Vv. Ter for their generous loan of equipment; ‘the crt Water, ‘Resources Center ‘provided — "valuable advice and ‘support. “Finally, we want to “thank the staff of ‘CRI, for all their, help. and. @ patience. — TABLE OF CONTENTS Page ABSTRACT cee me GED ee Jc roman Sane Se my SMO FEE TY Gee Ge Ge em Cee eae CaO: aN Gen Got Ges SER Le GD SO ene Gre Coe Ge One anme Gone rum iil ACKNOWLEDGEMENT me eee sm a me me, cr as ee mene sn nr iss sn on, wii TABLE OF CONTENTS vi LIST OF FIGURES oe dees ae cone Se are aoe eas ined Come Ge Gem WES Me CREE ne Gee ROS Cire iD oe Meee Oe Bae OY toe Vil LIST OF TABLES Soe eee ee oe ee Vill INTRODUCTION - METHODS AND ‘MATERIALS RESULTS. Oe Se eee ee Cie perme oe! come cht See es tne emcee eae Oa Ome Meee: Wem Sham We ee Gabe Smee Or tm coe cu Ge ee Gem ms ee es oe 14 DISCUSSION - 21 21 aL “Waterborne “Respiratory Pathogens: ‘b. Waterborne Gastrointestinal ‘Pathogens-~ 29 ee ‘Waterborne Disease’ and: Environmental. Health Considerations: "et ea aie come ie ee eo ees teres Se ein ec See “ines fume ane 34 CONCLUSIONS - ae are cee Se et ao ‘ame fame ee ep Yc Gt Sie ee eam ce ers ee ne es ne cee cake 38 REFERENCES. 4] APPENDICES et ee pcan AP 1 LIST OF FIGURES Page 1. Geographic Location Map for the Virgin’ Islands: ye es cea ee one ce ain ee er we ca cree wae ss te eae cs ne cos ee ane es acne sn ie 2. Diagram of the Main Trunk: Water Distribu- tion ‘System and Sample Stations: ae pee a naw ey i. ee ete eres dene Se 3. Genéralized Flow. ‘Diagram: ‘for Microbiological Se se Sy noe eh nny nn sd cee ee se es ed ee ee Samples~ 10 w“Vii- the: Virgin. Islands Seer erceettes | Quality. Control “Confirmation: Type Cultures Used seers are ana ee BS Ge setae paiacantes ieceireerm . Media Used In Isolation and Identification-- _ Source and Identification of Waterborne woe ; Bacteria. from the Virgin Petenep ccs Waterborne peatonella. Species” From A Comparison of: Two Indicators of ~ Contamination With Actual ‘Occurrence’ eee » Of Pathogens Humans: oe ee ae - Rtiology of waterborne. Disease Outbreaks In Le the United States, (1971- 1979 CUSTER Classification of Waterborne. Streptococci From. the V. Te and- fhe Diseases” They May Causerer Etiology of: Ve I. Waterborne ‘Bacterial - Diseases. Acquired Through the Respiratory _API- 20E Test , System asses Oleg ome Pract Si rehtorencminke nooner emerer anne nao oe - Rtdology | of V.I. Waterborne Bacteria cae ao Acquired ‘Through the Alimentary Tracto—----—— "APPENDIX. A. Classification. ‘of Medically | Important Waterborne Bacteria from the U.S. Virgin. Ts1ands ————--——— one “Appendix Be Procedures for Biochemical | Differentiation of Bacteria Using the -viii- - Pages aD 135 15. 25 26 AP 1 | significant “permanent populations. Th 1984, approximately INTRODUCTION ‘The U.S. Virgin Islands are a group of three, major islands, and. a7 smaller islands and cays clustered around the 18th paral lel. The. islands are about 1100 miles ae southeast of Miami (Pigure nD. 8 Only: the. three larger Yslands” of st. “Thomas, st. “gohn and st. croix support any 140, 000 ‘permanent "residents occupied the 60 odd square miles of land. In addition ‘beer 1. million persons a year visit the islands. ‘These visitors comprise a large ‘poten=". “tial | vector pool which could. transmit pathogens widely. - hocated in the. tradé-winds. zone the islands have” a high ve oe humidity but low rainfall. Annual rainfall varies from 20: Mo gnehes £0” 70, “inches per year ine different parts of the “. fslands. with ¢ this rainfall and ‘popllation: deena,” water’ “| becomes: a. ‘major commodity. the 4-5 million gallons of . public’ water per day. required comes — from three ‘sources: valnwater. catchment, groundwater, “and sea water. desalina- 7 thon. Of these sources. only. the desalinated public water oy is foutinely treated prior to. dist ribation. Even SO, due to an antiquated, "leaky distribution system, (see Figure 2) the: public water is. liable to “contamination before “reaching the publics All of the water, sources are cross — *SPURTSI UTATA 943 ZOz. dew uotqe007 oTyderboe9 + emnbta “sf .. epnqieg & : ae ‘Avasfauueg. “3S o- Pe IE! YS. Be Bjomop ‘ Aa epebouy NV390 OLLNVILV. ony oneng connected via unregulated commercial water “hauling truc ks, which carry” water from wells or the public. water, supply to cisterns in homes, businesses, and ‘public buildings: without cleaning the. trucks. “The. tor ad) Determine objectives of this research are. presence Or absence of common: human pathogens: in ‘the various water sources. of the v. Tey (2) to determine whether their presence has any. implications, (3) to evaluate whether the use. of Environmental Protection Agency : standards concerning coliforms is. adequate: for the various sources Of water in the Velo. ans _ ‘METHODS AND MATERIALS fs Water samples from each source (Figure 2) were . taken 7 -in four — liter sterile bottles. These bottles were kept at "ambient temperature, — transported directly to the lab, ~and » oe “analysis begun immediately. — “An aliquot was taken from the sample for conductance, turbidity and PH analysis, — zach | sample was filtered through HA and HC” membranes (Millipore, Mass.) in 5-10 and. 50 ml (duplicate) volumes. Where applicable, menbranes were preincubated on pads with oe "enrichment broths; — fOr 2- 3 hours on. laurel ‘tryptose “broth for total coliforms; Or 4 hours on. phenyl red lactose broth — at 38°C before ‘being | transferred to selective growth media, then ‘reincubated at an appropriate temperature (Canoy. and ‘gnudeen, - 1983). ‘quality control tests were run bys. a. untnoentated media for sterility, (@) using | American Type ‘culture | collection (arce), pure cultures with test samples, and (C) running : ‘simultaneous. biochemical tests on commercial media’ arid on Analytical Profile Index (API- -20E,_ 208 and = ‘Staph- Ident) I. De kits. “Anomalous tests were | “run again and also were| “queried ‘via. the API computer. “The” “APT, 208 system is a standardized, “miniaturized "version of. ‘conventional procedures for the “identification _ - Of Enterobacteriaceae ana other Gram-negative bacteria. rt ote a. ready-to-use, microtube (Bartoli, et aly 1972) system designed — ‘for the performance of 23 standard ‘biochemical ees from isolated colonies of bacteria on plating medium, Used - n. conjunction with the- API Profile Recognition — (Robertson, et al, 1976), it is’ intended to enable “laboratory. personnel to identify members of the family | oe Enterobacteriaceae and ~ “other Gram-negative bacteria . accurately _ and easily. — ‘the API system | has procedures for. game day and. 18-24 hour identification of “Bnterobacteri- ~ aceae as well as ‘1s- 24 or, 36- 48 hour identification of | : other. Gram-negative bacteria. the biochemical identifica- — tion of Salmonella arizona, Shigella, Escherichia coli, as well as Vibrio choler ae should be considered | a. presumptive - identification and confirmed agtplogiestiy. © specimen Collection and Processing . - . "Samples were ‘collected at 0700- 0800. at the » “gtations shown in Figure 2, ‘transported to the laboratory, immediate- oly after collection, and ‘processed beginning at 0900. Care “was used in proper selection. of adequate plating media and the ‘conditions of incubation. _ For the API. 208, “blood d agar -9-" ) suotzeis: etdues g, we4sdg: uoTANgnszysta yunay: us en auy jo werberc > -z armnbta SSE AE rane: sim. ames oR oe oe a . oe -7 : FW allolavHo a ae ‘plates. or. “MacConkey" Ss agar were streaked for. “initial i enrichment _ and isolation of colonies ‘to run through “the biochemical | testa. The APL system is standard for medical microbiology. | ‘Chemical and Physical: Principles “The ‘API 205. “system consists of microtubes containing — "© gghyarated® aubstrates, These: substrates “are reconstituted es by adding a bacterial suspension, incubated so that the or~— - ganisms | react with the contents of ‘the tubes, and read when eerie ‘the various indicator systems are affected by the “meta- bolites or “added reagents, generally after 18-24 hours in- “‘eubation < at 35- -37° es : "Components os | ‘The procedure for. each of the tubes _ are given in- = “Appendix B. “The: ONPG tube: contains an ingredient — that ee _ functions as an internal indicator. The ADH, LDC ’ ~ODC. and or URE tubes contain phenol red as. the indicator. The crt, os GLU, | MAN, INO, SOR, RHA, SAC, MEL, AMY and ARA tubes con- _ tain | charcoal _and the H. s tube contains iron salts as. "indicators. “the TDA, IND and VP tubes contain TO, oO . "indicator. — ALL. (of ‘the. tubes contain buffers, and tubes, with: the exception of the CIT, and URE tubes, con-_ | tain | peptone. All of. ‘these | "components retain “their "reactivity £ for 18 ‘months. ae Ident srveattee PE Brant ast “Identification of. the organism can “be: made either with ‘the. aid of API differential charts, or by using the compu-_ ter. Profile Recognition system, ‘when ‘differential charts are. used to determine the _— faentity of. cultures, — ‘the: aggregate reactions given by the _ostrains. should be ‘considered as. a whole. — Even when this is” ~ done, - however, the conclusions “reached. may differ “from laboratory to. laboratory. — | | wes | , "For rapid identification. of . Species and — biotype levels, the APL Profile Recognition system (PRS) , Computer) "Service cand Analytical Profile. Indexes. are ‘available. A “data ‘base. compiled” ‘from. strains of the ‘collections of deveral institutions as well as type and neotype cultures _ allows | very precise statistical calculation and thus ‘the identification of less | conmonly occurring microorganisms. “Quality Control cae Biochemicals "incorporated into the APL (20E system are: ae controlled by standard testing procedures prior. to. their usage in the systen. " Each lot is controlled with . a stock cultures for: sensitivity and specificity. Because. of the. use of lyophilizea cultures for quality control ‘procedures, several ‘transfers. were made prior bo the inoculation, ae : , oer recommends that quality, ‘control be- done. using the 18- 24 hour procedure. Same day results for ‘the recommended = ATCC cultures are available from Technical Services, _ API for: 1. Klebsiella pneumoniae. . ATCC 13883 2. Enterobacter cloacae ATCC 13047 3. Proteus vulgaris ATCC 13315 4. Pseudomonas aeruginosa ATCC (10145 - APT differential charts. have been derived ‘from type ‘and neotype cultures and strains from various “collections. ane 4 percentages obtained on API- may differ from percentages "appearing in ‘Published material based. on. macromethods. For example, the urease test found on API is a~ Ferguson — . (1948). formulation | which ae less sensitive Ose than ae Christensen’: Ss. "formulation which is used | widely in existing oe macrotechniques. - Reactions recorded as delayed “Positive with macromethods generally are “shown, as negative on the APL chart. ‘for Enterobacteriaceae since ‘this chart. is based on results | obtained exactly after. 5 or within 18- 24 hours . Of incubation depending on the. ‘coordinate parallel standard tests being carried on at the ‘same times * Figure 3. Generalized Flow Diagram of Microbiological ‘Samples. . . L “Three Dilutions: vast 50 ml to 100 ml on ae ate eee ea oe ‘Membrane to Enrichment Broth | Quality Control == =. 2 Or 4 hr. iae" ome . Culture Organisn . :-¢g_, For Media Perfor- . : for sterility cheek. (Confirmation Tests Set A). American Type -flance plus blanks & , "Key n Selective media a ‘aacond "x25" pifferential Media - feonetemtson Tests set 2B) ae Bative. (end) & Negative Lg : Qu wality ‘Control APT Test. ‘Strir Media Sterilit : -mhujca1/Bioctentcal . Characterization Media APIS ~Test: ; me = The ‘first “quality control. effort is made by setting — aside 7 1s of ‘the | containers with “uninoculated media, which - is watchea for contamination. A second - “quality — "assurance _ “measure involved inoculation of each nedia with pare | culture bacteria from the American. ‘type, Culture Col- 2 lection z (amec) “(Table 1) and observing _ their, relative reactions. The final confirmation was a comparison of Ana=_ lytical Profile Index (API). test strips with “standard PH, 1984), characterization medias. Aree 4 Table. i. “quality Control Confirmation: Type Cultures Used Difco Bactrol Diske | (Lot + ice 32- 2 and Lot # 1656- 32-1) ATCC 8100 - Serratia marcescens "ATCC #13315 - Proteus vulgaris ATCC #27853 - Pseudomonas aeruginosa - ATCC # 23355 - Enterobacter cloacae _ ame # 13048 - Enterobacter aerogens 25922 -— Escherichia coli. 8090 ~ Citrobacter freundii - apcc #19606 - Acinetobacter calcoaceticus; var. anitratus ATCC 14028 - salmonella typhimurium 19615 - Streptococcus pyogenes BTCC. # $ # # $ # . "—amec #13983 - Klebsiella pneumonia ; : $ $ $ # 25923 - s staphylococcus aureus. - ay / ance # 12228 - § Staphylococcus | epidermitis oe ‘Table 2. “Media used In Isolation and. ‘Tdentification _—-mEndo-Agar-LEs ‘MFC-Agar a mStaphylococcus Broth. - mBrilliant Green Broth mBismuth Sulfite Broth. -mTetrathionate Broth Base ‘Phenol Red Lactose. Broth © “Plate Count . Agar Brain-Heart Infusion Broth -. Brain-Heart Infusion Agar 9. : . Brain-Heart Infusion Agar: w/5% blood - Nutrient Broth: — ; “Simmons Citrate Agar MR-VP Medium ~ Tryptone: Urea Disk ae , Lauryl Tryptose ‘Phosphate | Broth Brilliant. Green Bile Broth 28 “EC Broth. © fel kes (Oxidation-Fermentation Glucose ‘Medium Me - Levine EMB Agar... ee ae Bismuth Sulfite Agar ~ 8/S Agar: Pseudomonas. Isolation. Agar. Motility GI Medium — - Rabbit Plasma. w/EDTA XLD Agar oo , -. Sellers Agar - Hectoen Agar . _ MacConkey Agar ee a _ Triple Sugar Iron Agar - Yersinia. Selective Agar - DNase_ Test Agar w/Methyl Green -. Decarboxylase Base (Moelter) w/tysine. _ Arginine & “Ornithine | Nutrient Agar. : KCN Broth Base. -Phenylalamine Agar . -- Bile Esculin Azide Agac: KF- St reptococcus-Broth surskqoadexig - _oTx0} “uTpTooyney OSES aseutyoideris teseptuointedy | _-. .sutsdpoqderyg . «ded prose’ stuoinpeAy ty ayeapAyoqtes H dnosy aqerpAyoqred y dnoip » xeTdwo> pte outue -oyerpAyoqred dno, _ eyexpAyogzes *9 dnosg -oqieo y dnoig efns . eudly. eydty — eudty 210g e1O8g Bqeg. _ SATIRATTES: ~ _. STPTUESTAbe - “*ej0q ‘eydte 8q Aeu yey) mn990 T25000Ader3s (I afidkin FO soTDdS’ TOYIO WY) rq ae El ui ie whed an bs - % STAOG ‘$) 0 TopeD07SAUe=UON oy (STTBDORF : *S) Toosoremay Oe Sows “FV Seseostq “TeOTUTTO. "~~ Sgonpolg re[NTTOIeLIXg “Syonporg | _ FRTNTTSO © se padg OO aoig oo ppergzoouey © Sane da Kou SoseosIa aia PUE “TA ot uous "PoddeadaLas FuLOG LeIeN Jo VOIIEDLETSSEL _ Disease Table 6. Etiology of Vv. I. waterborne Bacterial ‘Diseases aes : Aeaeired Throsah the Respiratory Tract ae Other Possible ~ Causative Entry Routes_ — Organism “upper Respiratory Infections. Pharyngitis tage (children) — Streptococcus (group A) o stre tocéceus pneumoniae og Streptococcus _ (group A). Sta taphylococcus — aureus enteric bacilli. Gram-negative. Bacterial Pneumonias Other bacterial. * pneumonias. eran ' Legionnaires' _ disease. - . heaichelia Streptococcus pyogenes _,Staphyloccus “aureus - Klebsiella ~~ pneumoniae ‘Unknown — Bneumonphila - ot Strep (sore) _ "atypical. no ee ycobacterium - Spp. (mot Me Eubereuopis) oc “Table 6. con't | - Clinical Disease: “Causative 7 Organiam «= __ Streptococeal Diseases throat — “Scarlet. fever. eg ‘sequelae of group - "A" strep infec- tions: = - Rheumatic Fever. “Acute hemorrhagic” "Streptococcus “possible . Entry Routes —_. PYOGENES Streptococcus _ ae “pyogenes — “glonerulonephri~ tis oe “Tuberculosis. peer a Tuberculosis — ee mycobacteria v0. Waterborne Gastrointestinal Pathogens (See. Table 7. ; for etiology) ws salmonella (Salmonellosis) The genus _ Salmonella occurs: Pies of man and — -. a number of birds and animals. _ Tt is normally spread via _ water or food contamination. The organism prefers. nutrient orton nedia and. temperatures of less. than 40°C | While there are. several” hundred serotypes known only” some -20=30 are — and of those only about A3- 15 normally cause ‘disease gn homanne c= The “typhoid. Fever" organism Ss. ‘typhi ‘is. the only epsaie® dtpenne epeeisic for man. ‘Salmonellosis (including typhoid) AR one time over 1800 ‘serotypes: of: ‘Salmonella were. con oeigered “separate “species. Now this has been reduced to” _ about” 200: and probably soon. this will reduce to. one or. two "serologically. highly diverse" species. while” only ‘Ss. “typhi is specific for many: humans, perhaps ‘most (Of the others, are pathogenic to “some ‘degree, “Gastroenteritis is. the generic term for ‘Salmonella good ‘poisioning. _ Various “Salmonella species: may” be in- volved” -and the source is. usually. water or food. an mary. cases” “contamination comes from ‘a non-symptomat ic person _ te ("the carrier") (Or from the animal which produced the. food, as often occurs with eggs. as “Salmonellias were found in a “well on st. ‘Croix, during a. previous: study (canoy, Knudsen Btioloay. of. V.I. Waterborne Table Te Bacteria » Acquired: Through the. Alimentary: Tract _ Clinical RAgease - Causitive ae Organism.” 2 “giles bemeinie Entry Routes. “Reate Gastrointestinal ae infect ione -- salmonellosis — Piso (bacillary — ae dysentery) - "Gastroenteritis : “Salmonella spp. | oe _..(many sterotypes) = — . Shigella spp. . "-Enteropathogenic oe Es coli ‘Campylobacter Yersinia’ ve enterocoli tica Sexual “Placental Water. - SPP. _.. Respiratory? — Water? “Bacillus. cereus fake Epidemic diarrhea Seo imureertes.. ve: “Enteropathogenic : Escherichia coli _ _ Biaamaas Coes by Ingestion of Preformed Toxin "Staphylococcal ambpx icabion 7 Staphylococcus aureus “chterotoxin Noes ° Table 7.0 —. Clinical eo . Causative. i apeage ce a Organism, Other Possible. Entry. Routes — Infectious. systematic Diseases” Typhoid Fever Salmonella 2 vey? typhi = Salmonellosis _ : alameda top Galmonella) "Salmonella cholerae~ . septicemia’ fs esuis — | re “and Garcia, 1985) but were “not definitely identifica during this study of water in. ‘the Virgin Islands. a8 Due to the generally” nigh number of Salmonella necessary ‘to cause - an. i. infection ‘and the probability, of being | "masked" by other "bacteria, a lack of disease or. colony isolates does net” _nectasantly: mean they are. absent. “The genus salmonella was isolated from a cistern. in . ste ae. in 1983 (Caney and > Rnudsen, 1983). oe ‘Shortly Fan _ thereafter a. few. isolated cases: of typhoid occurred on. Steg Croix. A study of wells in the Vv. Te in 1984 confirmed — ‘Salmonella SPP+ from two. wells on. ‘st. “John and one on St. — Croix. In 1985, the 3rd largest U. Se outbreak of. ‘typhoid occurred on. st. croix ‘totaling 66 cases hospitalized. ‘Considering, the: low level of Salmonella, even. ‘in known _ contaminated areas, detection is. snot. easy in any case. To. oe find ‘positive ‘identification ine four ‘sites was in itself “cause for some. ‘concern. — No. attempt was. made to. . identify a “the « organisms to species, which “would | have “required “serology and/or immuno-fluoro ‘techniques presently “beyond the laboratory. - | Shigellosis is an intestinal. disease — of varying severity ranging from simple ‘dysentary BO / cholera- like. symptoms. Shigella species are responsible for about 308 of the "summer dysentery" outbreaks in the 7 8. 8. ‘In ‘the. v. Ss. and “Europe shigell: La related fatanities | percent. are only about one. > percent. ; In tropical areas with poor” “health care; ‘fatalities — vin. epidemics may reach twenty - Salmone ella an and ‘Shigella spp. are not detectable using ce ihe standard indicator tests. — only. lactose | : getménting °° . "bacteria are detected in ‘standard water. tests; and except for. a few genera these are non-pathogenic. o Becherichia coli, Enteropathogenic varieties (acute | Gastroentritis) i | a are Bs gol is a normal inhabitant of the ‘human colon. | Pees as. such. it has. become the standard “indicator organism — oes for human fecal pollution. “Many scientists now. criticize this on ‘the basis ety "he “gold is not @ pathogen. Stn “actuality about. O. 5 -.. 08 of the U.S. children under 5 years of age: and 2. 2 - 3. 388 of the children in ‘developing = countries are stricken » with E Ee goli gastroenteritis (Wo . annual report, 1978). a The symptons ¢ of the disease are a profuse. watery aiarrhea, with little mucous. or ‘blood, fake : Bear Prostration and» ‘dehydration. ‘The organism apparently ee ‘excretes: an. “enterotoxin and also. penetrates the » colonic “epithelia. — with the. advent of modern antibiotics and in- _—-tervencus feeding, ‘the disease in the “developed countries eo ha ‘become only” an ) unpleasant nuisance, In developing petedey countries, and in poverty ¢ areas of. the U. Se it. is a serious, ee often fatal, disease of young children. - the isolation, of hydrogen sulfide 088) ‘producing i re E.coli from wells (Canoy, ‘Knudsen, Garcia, 1985). and from public” “drinking water. (this: study) indicates a potential — pathogenic Es goli. Due ‘to ‘the biochemistry, of this - variety | at. is not detected | on. standard” confirmed “fecal “coliform tests. S The presence of these bacteria in’ gbhe. water. is, especially for ‘infants and “medically compromised patients, of ‘concern. _ “Waterborne Disease ‘and Environmental “Health oe Considerations : potential dispersal routes: “for waterborne pathogens "Human sate. | . mus gay a "potable. Water Distribution system oe “Water: Transport | ‘Trucks, ~ gewage - -Rain/Wind . Human vectors. (as voice to “general sewage) include | “persons 4 who handle food or water or those. who» deliberately ‘spit or. excrete into. and around cisterns and wells. whis) S category» -also_ includes: ‘persons ans charge of wells, | ae trucks, and | barges who, do not. clean, or. close them properly. os ee - At ‘this time (ages), there. is no satisfactory est imate i a to. the. importance | of this vector. — In a, recent typhoid " epidenic, statements were made, but not substantiated, that "food handlers - were ‘the ‘source. — other reports. said that Q feces. “deposited around or. in the cisterns, were the “source “Sor blamed cross” contamination between “water and sewer | a pipes. ‘The importance of this vector needs to be addressed even though it is very time ‘consuming. The” ‘potable water distribution system has been. blamed — often for “allowing cross-contamination with sewage. Again “there is no good est imate of ‘Ene: magnitude of this problem. "However in the Present ‘study | six. samples were taken. along. . the | distribution dine. — The closest sample to. the. storage tanks was number 262 ‘in ‘Subbase and had two. ay potential ereeeen "pathogens. After ‘this point, one pipe goes. to Veteran’ SS "Drive West and one. to. veteran’ Ss. ‘Drive East and the town of. oes “charlotte Amalie. ‘The Veteran! 's Drive West ‘sample (267 Doo. a nine 3) pathogens, the. East Veteran! Ss. ‘Drive (264). had (2) pathogens and (261) east of town (towards: end of Line) had ‘thirteen (a3) pathogens. — Another sample, on a branch Of the system, was from. the airport and had | eleven. ay potential pathogens. ae Pee From the ‘limited number. of samples it appears that cin = gece: the: farther from the, storage and ‘treatment: site, the MNES BB oe extremely £luorescent dye ‘such as "greater numbers of total bacteria and potential ‘pathogens. “This. would: imply some. sort of cross contamination. — As critical as this factor is, a great | deal of importance should be placed on: dete mining. . the | locale — of oe contamination... ‘This | could be done in 100. samples using — “both bacterial tests at various” ‘Rhodamine B B injected into. the | sewage “lines. “It is possible that area ‘treatment — oe “should be installed o ‘other precautions taken. " Water transport — trucks are. he vate to. areas not served ‘by the public. distribution — sys- tem, Some of these trucks are owned or leased - by the ‘Housing Authority or: Public Works, however, most | are private contractors. “They all have a potential for conta~ mination | from polluted wells, public. cisterns, | or illegal : withdrawals fron the: distribution system. since there is oints in the lines, and an used to deliver. ‘drinking = “no, control on servicing, — cleansing “monitoring, these nh - trucks are potential “hasaraay “The: owners will not. permit cee - “sampling by private researchers and SO. any evaluative study wil have to have government backing. Furthermore, because of the. unique environment, “any simple. coliform test “would be meaningless. “The tests required would be a ‘Total. Plate Count plus ‘some “other — indicator, “perhaps: - fecal ; “ treptococcus, Clostridium. Aeromonas, or ‘Pseudomonas. ‘Sewage contamination’ has often been blamed ‘for lowered water quality 3 ‘in. cisterns, wells, and. the public system, The ‘type and number of bacteria found in this series of x studies _ (Rhineheart, et aly 1983; ‘Canoy, Knudsen and: @areia,. 1985) | confirm “sewage contamination but do- not "pinpoint: a single source of contamination. There are three ~ major potential sources; cross contamination “between water and sewage pipes, surface and subsurface entry from septic 7 tanks, and. runoff carrying surface wastes. No factorial es has: ever been done to evaluate these three routes. , ‘Animal vectors. include cattle, pets, wild birds, rats, es : A wobgooaeas and “reptiles or amphibians. These animals” “deposit feces “around poorly - sealed wells and cracked _ cisterns. At times not only the feces of smaller animals _ but smaller animals themselves may enter, directly into — open. cisterns or wells. “These animals carry” pathogenic. -Salmonellas, _ Strept ococcus: and atypical | tuberculosis: a8. well as a an - variety of Aeromonas and ‘Pseudomonas. Rain and wd nd “play can _ unknown tele. in barry ing, garee. pathogens into wells” and "cisterns. Road ‘dust is” . constantly being swirled up into the air to be added to water foamites: from wastes: and sewage aerators: as. well as. 23 7- dust | ‘borne on. upper winds: from as s far away as” Saharan. Africa. — This mixture can ‘then be rained out on. water ~ catchment surfaces locally. . _ CONCLUSIONS “this research has. ‘shown, ‘that ‘the private and public ~ potable water systems in the Virgin Islands are frequently contaminated with potentially hazardous bacteria. . In the “present study, and ‘two preceeding efforts; 125 cisterns, 26 wells, and 6 public water. outlets were tested. Even, though the tests. were not exhaustive for. all possible “pathogens only ‘two wells, ‘four cisterns, and. no public water outlets _ were” found to. be free of ‘the present EPA standard indicator ‘bacteria, the implications of ‘this, in terms: of basic an human health and_ high risk to infants, elderly, and medically compromised persons are dramatic. _ Special 1 moni- 3 toring and prophylaxis. “for, the high risk groups: are a “critical need. Furthermore the large influx of. tourists and “foreign immigrants in the “Vale. complicate the water - pathogen ‘situation by the introduction. of organisms and by providing © . large mobile host “reservoir because “those | east tues ) from many: ‘greap., where typhoid, cholera, “yellow fever, and other “ Mexotic™ pathogens _ still are oe active. When introduced here, transients going to the Us S. “mainland or ‘Europe could easily transfer the disease before oP faage Fe Me even identifiea like the controlled outbreaks of typhoid in -1ge4- -85. “This: “would be nothing short of an ‘economic, as well as. a 1 public health, crisis. The. majority of the pathogens found cause. one of three "general maladies: ly “Skin or wound | “septicemia, 2). "Respiratory and Genitourinary tract infections (particu- larly “Of women), 3). Gastro-intestinal infections, Less 4 : common nee. the pee atypical “bacterial pneumonias — and. tuberculosis. — ‘These bacteria are not. generally (but can oo become) | primary pathogens and so. usually become critical in already — weakened. hosts” or. cause: “chronic low level | debsnseating dcasea | a | bends “Current water: standards in the: ve I. are those wee in the U. Se for, a ‘different geography, popula- oe “tion, and water system. Neither the tests, ‘monitoring, or. oe “indicator organisms appear appropriate to the local situa oo tton. ‘These are. particularly, questionable in view of the | ee - diverse ‘sources — of water, — dissolved. organics, 2g heat, turbidity, ee and storage times Of water. Studies are - required | to provide data to. ‘develop more appropriate ‘tests wee and to. “recommend to the lawmakers. more appropriate “local “regulations for water systems: including wells, trucks, and public cisterns, “Strict” ‘regulation. of water Saeedithen and treatment, septic tanks, and ‘sewage is’ ‘required, _In addition to the physical measures called for, a massive educational effort is necessary on the part of — “" gchools, government, and civic organizations to create an informed and active public relative to water quality. 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Enterobacteriaceae_ Motile or. nonmetile differentiated by. carbohydrate fermentations and > antigenic properties; some. conmen- - galistic. in intestinal tract with ",. Genus IV: Salmonella pathogenic potential for extra-. eee intestinal tissues; some are paths ae _ ogenic” parasites. acquired ‘through. “alimentary: route Genus I: "Escherichia os os Epidemic diarrhea of: , are Es coli poe ee infants; urinary: tract pene and wound infections; ane "turista™ S$, typhi ee fever. . Be enteritidis paige he Gastroenteritis, para~ : ; o-.. .typhoid, and enteric - fevers: (many. varieties — a of this species may - es S@ause these diseases) _ Ss cholerawsuis Septicemia, muleipie ES 8 eee _ abscesses. “Genus Vi Shigella g. Se : boise eek — boydi “ pveenéeis: . devere diarrheic. Ss. sonnei isease Genus VI: Klebsiella foe oe os ey AED 2a a pneumoniae SS. pneumonia: os, te Fey ooo Urinary: tract infec- a . tions eas os Genus 1: : Table 8 Con ' t Classification —— Disease Enterobacter E. cloagae. E. aerogenes... Es agglomerans — © Genus VII: ~ ~- Genus XII in . family Entero- -backeriaceae) Serratia. Genus IX: § ee ET Se marcescens. - Proteus P. vulgaris. ~ Genus X: Ba alrablis versinia eee : Dog Egy enterocolitica Genus XII: Erwina ake ty ta herbicola oe ‘plant: pathos -- gen) also class- pases as Entero-. “bacter scene “pamily - II. VIBRIONACEAE “Genus II: Aeromonas ~ Genera of uncertain Family Genus “Legionella - Le: pneumophila — a ee ‘apeta teuleed veda: oe Normal intestinal tract, “may cause urinary. tract a infections ~~ -.. Has caused systematic (see Erwina’ ils infection when. intro- ~- duced in contaminated. -- solutions. injected in- _ travenously — “Urinary and: respiratory - tract infections _. oo “Urinary tract and wound. infections; infant | diarrhea . Intestinal tract of oe animals and humans, may —. Gause: diarrhea, 5 ae mesenteric” lymphadeni- tis, abscesses, | iInenin- . —gitis “Has carved. ‘Byatemic ' infection when intro- duced in- contaminated ee . solutions injected intravenously," fe “Marine: environment, seafoods, may cause . acute enteritis cota wd ‘Associated with sys-_ -- temic and intestinal _ infections — ee a “Legionnaire' s. @isease: (mild. to ‘Severe ‘pneumo~ _ nia). ae o Mable 8 Con't ~ Classification — . Disease ~ “.C. Group: Gram-positive cocci, aerobic and/or. facultatively © -. anaerobic, some ‘strictly ~ ahaerebie : ane ~pamily I.MICROCOCCACEAE . Arranged in. clusters. or pack-- ets, saprophytic or parasitic, identified by coagulase, hemo-. =. — -- lysin, and pigment production _ Staphylococcus Genus II: | ay pe oo S. aureus -. S. epidermidis as saprophvticus ‘skin abscesses, impetigo —s. wound infections, pneumo-_ nia and other systemic . infections, enterotoxin- — _ producing strains cause . _.food poisoning, exfoli- - - ative toxin causes - - - -- "scalded skin". syndrome infants, other toxins. ae cause toxic shock | syn- drome.) Normal commensal of skin “but may cause. local. or systemic infections if-. ~.. introduced by contami-9 “nated sutures or instru- ments coed . ‘Associated with urinary” tract infection in. young: women Sasssisiontion ‘Disease | amily. Il. " STREPTOCOCCACEAE . Changing cocci, .commensalistic and/or pathogenic : Genus I: Streptococcus | ee oo Se Pheumoniae. a Se) pyogenes. - (usually beta-_ ~ hemolytic) Ss. faecalis” _ (enterococei") viridans’ group. . (alpha hemolytic “species that pro-. . duce greening of. blood agar); in- cludes: following | species: | Se mitis . -§, galwarius. — $. mutans Pneumonia, other sys~ temic infections — “Scarlet. fever, sore throat, erysipelas,. impetigo, postpartum. endometritis, wound. | ‘and burn infections, aon and other serious diseases . Normal intestinal tract, os . sometimes cause res- _ piratory and urinary tract infections, .—- ‘infective endocarditis. Intestinal tract of cows, -and other animals, = ‘causes: human endocar~ itis “Mastitis in cows, in i -.. humans, neonatal sep- ‘ticima and meningitis . - (infected mothers may also have sérious ae systematic disease) Sesh Normal respiratory. tract; — chronic mucosal. in-. fections: infective endocarditis “Normal ‘veapiratory tract; may cause endocarditis, - dental caries: Same ‘as above _ Major cause of. dental caries; may cause . endocarditis. : Tmpet igo shnefngitle