Waterborne Toxoplasmosis, Brazil, from Field to Gene.
Water was the suspected vehicle of Toxoplasma gondii dissemination in a toxoplasmosis Toxoplasmosis Definition
Toxoplasmosis is an infectious disease caused by the one-celled protozoan parasite Toxoplasma gondii. Although most individuals do not experience any symptoms, the disease can be very serious, and even fatal, in outbreak in Brazil. A case-control study and geographic mapping of cases were performed. T. gondii was isolated directly from the implicated im·pli·cate
tr.v. im·pli·cat·ed, im·pli·cat·ing, im·pli·cates
1. To involve or connect intimately or incriminatingly: evidence that implicates others in the plot.
2. water and genotyped as SAG 2 type I.
Water has been considered an important vehicle for disseminating human toxoplasmosis in outbreaks (1,2) and in endemic settings in Brazil (3). We investigated a large toxoplasmosis outbreak in which the exposure to known sources of Toxoplasma gondii infection was assessed. We found that unfiltered Please wikify (format) this article or section as suggested in the Guide to layout and the Manual of Style.
Remove this template after wikifying. This article has been tagged since , municipally treated water was the epidemiologically implicated source of infection for this outbreak. Isolation, polymerase chain reaction polymerase chain reaction (pŏl`ĭmərās') (PCR), laboratory process in which a particular DNA segment from a mixture of DNA chains is rapidly replicated, producing a large, readily analyzed sample of a piece of DNA; the process is (PCR PCR polymerase chain reaction.
polymerase chain reaction
Polymerase chain reaction (PCR) ) detection, and genotyping of T. gondii from the implicated water source were demonstrated.
In November 2001, in Santa Isabel do Ivai, (southern state of Paranfi), a local physician requested serologic tests to diagnose dengue dengue
or breakbone fever or dandy fever
Infectious, disabling mosquito-borne fever. Other symptoms include extreme joint pain and stiffness, intense pain behind the eyes, a return of fever after brief pause, and a characteristic rash. , mononucleosis mononucleosis /mono·nu·cle·o·sis/ (-noo?kle-o´sis) excess of mononuclear leukocytes (monocytes) in the blood.
chronic mononucleosis chronic fatigue syndrome. , cytomegalovirus infection, hepatitis, and toxoplasmosis in 2 persons in whom fever, headache, and myalgias had developed. Positive results were obtained for anti-T, gondii-immunoglobulin M (IgM) and IgG only. Through the end of 2001,294 similar cases, which were serologically confirmed as toxoplasmosis, were reported to health authorities in the same area.
The outbreak peaked between November 2001 and January 2002 (Figure 1). Symptoms were reported by 155 persons; the main symptoms were headache (n = 135), fever (n = 128), malaise (n = 128), myalgia myalgia /my·al·gia/ (mi-al´jah) muscular pain.myal´gic
epidemic myalgia see under pleurodynia.
n. (n = 124), lymphadenitis Lymphadenitis Definition
Lymphadenitis is the inflammation of a lymph node. It is often a complication of a bacterial infection of a wound, although it can also be caused by viruses or other disease agents. (n = 117), anorexia (n = 107), arthralgia arthralgia /ar·thral·gia/ (ahr-thral´jah) pain in a joint.
Severe pain in a joint. Also called arthrodynia. (n = 95), night sweats (n = 83), vomiting (n - 60), and rash (n = 11). The duration and magnitude of the epidemic curve could have been influenced by the intensity of media reporting at specific times, which led to people seeking toxoplasmosis testing.
[FIGURE 1 OMITTED]
Case-patients were located by active contacting of and passive reporting from local physicians, and media campaigns (television, radio, and newspapers). A matched case-control study was conducted from January 15 to February 2, 2002. Acute cases were defined by standard serologic se·rol·o·gy
n. pl. se·rol·o·gies
1. The science that deals with the properties and reactions of serums, especially blood serum.
2. criteria (4) and were selected from a list of volunteers. A total of 2,884 of 6,771 persons living in the urban area of the city volunteered to be serologically tested. A total of 426 (11.5%) persons had anti-T, gondii IgM and IgG antibodies; 1,255 (51%) were positive only for IgG antibodies. Of 426 persons who had anti-T, gondii IgM and lgG antibodies, 176 met the case definition; of these, 156 (89%) participated in the case-control study. Sex and age matched controls ([+ or -] 5 years, n=220) were selected from the same group of volunteer who were asymptomatic and seronegative seronegative /se·ro·neg·a·tive/ (-neg´ah-tiv) showing negative results on serological examination; showing a lack of antibody.
adj. for T. gondii.
Serum samples from case-patients and controls were tested for anti T. gondii IgM and IgG antibodies by the Central Laboratory of the Paranfi State by using 3 different commercially available enzyme-linked immunosorbent assays (ELISAs) because it was not possible for a single vendor to provide the number of required kits. Fifty percent of the case serum samples (78 samples of 156 participants) were randomly retested in a toxoplasmosis serology Serology
The division of biological science concerned with antigen-antibody reactions in serum. It properly encompasses any of these reactions, but is often used in a limited sense to denote laboratory diagnostic tests, especially for syphilis. reference laboratory, Laboratory of Protozoology protozoology /pro·to·zo·ol·o·gy/ (-zo-ol´ah-je) the study of protozoa.
The biological study of protozoa.
the scientific study of protozoa. at the Tropical Medicine Institute of Sgo Paulo. Five (6.4%) IgM- and IgG-positive serum samples, tested previously with 1 of the commercial kits, showed very low IgG avidity avidity /avid·i·ty/ (ah-vid´i-te)
1. the strength of an acid or base.
2. in immunology, an imprecise measure of the strength of antigen-antibody binding based on the rate at which the complex is formed. Cf. when tested by this laboratory. All the other serum sample test results were confirmed by testing conducted in this laboratory.
Of the 156 participants, 138 (88%) lived in the area served by reservoir A and 17 individuals lived in area served by reservoir B (Figure 2); 1 person had a private well. Table 1 shows the univariate analysis results and Table 2 shows the multivariate analysis results. Case-patients were significantly more likely than controls to drink water supplied by municipal reservoir A than reservoir B, as well as to eat commercial ice cream than not. The 4 case-patients that reported not drinking water from reservoir A, however, reported eating ice cream. The frequency of eating ice cream among the persons who drank water from the reservoir A was 32%.
[FIGURE 2 OMITTED]
The environmental investigation included mapping the city water supply system which is served by 2 municipal tank reservoirs (reservoir A and reservoir B) that both receive water from underground, protected deep wells. Both reservoirs are tanks with 150,000 L storage capacity. Case distribution showed a clustering in the central area served by reservoir A (Figure 2).
Because the environmental investigations and the case-control study started in parallel on January 9, 2002, and the outbreak had peaked (Figure 1), the chances of detecting parasites in the municipally distributed water were theoretically low. To increase the chances of detecting the parasite in water, household tanks that had water that had been distributed during the outbreak peak were identified. These tanks could be investigated in municipal schools that stopped water use due to summer vacations from December 17, 2001, to the end of January 2002. Despite the risk from eating ice cream (Table 2), no ice cream made during the outbreak period was available for laboratory testing. The ice cream was prepared locally in small batches with water from reservoir A.
We identified 4 schools that had water in their household tanks that had been distributed by reservoir A during the peak of the outbreak. Approximately 4,650 L of water collected from these tanks was filtered through 56 fluoropore membrane filters (Millipore Billerica, MA, USA). We retrieved 19 liters of water concentrated to 60 mL by centrifugation Centrifugation
A mechanical method of separating immiscible liquids or solids from liquids by the application of centrifugal force. This force can be very great, and separations which proceed slowly by gravity can be speeded up enormously in centrifugal (600 x g 30 min 4[degrees]C). The membrane filters were divided into 3 equal sets. One set remained in Brazil (Universidade Estadual do Norte Fluminense Darcy Ribeiro) for bioassays in T. gondii-seronegative chickens and further genotyping. One set was sent to the US Department of Agriculture for bioassays in T. gondii-seronegative pigs and cats, and 1 was sent to the Centers for Disease Control and Prevention Centers for Disease Control and Prevention (CDC), agency of the U.S. Public Health Service since 1973, with headquarters in Atlanta; it was established in 1946 as the Communicable Disease Center. for PCR analysis. Chickens and pigs were fed with membrane filters and their serum samples tested by ELISA ELISA (e-li´sah) Enzyme-Linked Immuno-Sorbent Assay; any enzyme immunoassay using an enzyme-labeled immunoreactant and an immunosorbent.
n. and or modified agglutination test (5) until seroconversion seroconversion /se·ro·con·ver·sion/ (-con-ver´zhun) the change of a seronegative test from negative to positive, indicating the development of antibodies in response to immunization or infection. . The seropositive seropositive /se·ro·pos·i·tive/ (-poz´i-tiv) showing positive results on serological examination; showing a high level of antibody.
adj. animal organs were examined for T. gondii (6). Control animals were fed with noncontaminated membrane filters. Water samples from the 4 schools' household tanks were positive for T. gondii by at least 1 assay method. Parasites were found in the lungs of mice injected with brain and heart tissue of seropositive chickens. Cats fed pig tissues shed T. gondii oocysts after 4-5 days. Oocysts from cat feces were injected into mice, which died of acute toxoplasmosis. Viable T. gondii was recovered in mice after subpassage as verified by optical microscopy. The nested amplification of SAG 2 followed by restriction fragment length polymorphism restriction fragment length polymorphism
n. Abbr. RFLP
Intraspecies variations in the length of DNA fragments generated by the action of restriction enzymes and caused by mutations that alter the sites at which these enzymes act, changing identified type I T. gondii from chickens and pigs (7).
DNA extraction from fluoropore membranes was performed with the FastDNA extraction method (Qbiogene, Irvine, CA, USA), by using a procedure previously published (8), and PCR was performed on extracted DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. by using primers Toxo B22 and B23 (9). PCR from DNA extracted directly from the fluoropore membranes was tested blindly by 2 persons on 3 aliquots extracted individually from each membrane filter. The correct size fragment of 115 bp from B1 T. gondii gene was amplified from each DNA aliquot aliquot (al-ee-kwoh) adj. a definite fractional share, usually applied when dividing and distributing a dead person's estate or trust assets. (See: share) extracted from membranes used to process water from 3 of the implicated tanks.
Our investigation determined that this toxoplasmosis outbreak was associated with consumption of contaminated water, or ice cream prepared with contaminated water, during the outbreak peak. The main factor leading to contamination of reservoir A was the vulnerability to infiltration due to its precarious state of conservation. We propose that reservoir A was contaminated with T. gondii oocysts because 1) a female cat living in the reservoir A area delivered 3 kittens in early October 2001; 2) the kittens lived on the top of the tank reservoir; and 3) the kittens were most likely weaned by the first week of November. However, it was not possible to confirm T. gondii in the kittens because we were not able to catch them. The reservoir shelter roof tiles were removed and not replaced until the end of heavy summer rains. From November 4 to December 12, the daily rainfall varied from 27 mm to 72 mm. Reservoir A, constructed in the 1940s, had cracks that were unprotected from rain water, which were likely contaminated with cat feces. These factors could have been enhanced by the lack of filtration and flocculation flocculation /floc·cu·la·tion/ (flok?u-la´shun) a colloid phenomenon in which the disperse phase separates in discrete, usually visible, particles rather than congealing into a continuous mass, as in coagulation. processes as part of the water treatment. Additionally, the level of chlorination chlorination Public health Addition of chlorinated compounds to drinking water as disinfectants. Cf Ozonation. used to treat water in municipal systems is inadequate to eliminate T gondii oocysts (10).
Of the 408 case-patients examined for ophthalmologic conditions through February of 2002 who were Toxoplasma Toxoplasma /Toxo·plas·ma/ (tok?so-plaz´mah) a genus of sporozoa that are intracellular parasites of many organs and tissues of birds and mammals, including humans. T. gon´dii is the etiologic agent of toxoplasmosis. IgM and IgG positive, 10% had ocular lesions; however, only 4.4% had necrotizing necrotizing /nec·ro·tiz·ing/ (nek´ro-tiz?ing) causing necrosis.
Causing the death of a specific area of tissue. Human bites frequently cause necrotizing infections. retinal lesions (11). The frequency of symptoms observed in this study may be associated with the dose and virulence of organisms ingested since parasites of genotype I, which are of high virulence (12,13), were isolated from the water implicated in the outbreak. These data are consistent with other studies also showing SAG-2 type I parasites isolated from the environment from different geographic areas in Brazil (14), including in the outbreak area (15). Demonstration of the parasite in the outbreak implicated water was decisive in the closing of reservoir A and the construction of a new municipal reservoir.
We thank Jarbas Barbosa da Silva Junior, Maria Regina Fernandes de Oliveira, Ana Eliza Mazzotini, Ant6nio Moreno, Marcos Cardoso Marques Marques may refer to:
throws over lover for another. [Fr. Lit.: Carmen; Fr. Opera: Bizet, Carmen, Westerman, 189–190]
See : Faithlessness
the cards repeatedly spell her death. [Fr. Lucia Muricy, Wildo Navegantes de Araujo, Alessandra Araujo Siqueira, Roberto Mello Dusi, Heitor Franco de Andrade Jr, Enrique Medina Acosta, Erica dos Santos Carvalho, Fernanda Santos Nascimento, Patricia Silva Santos, Daniele Seipel da Silva, Juliana Salgado Viana, Fernando C6sar Lopes, James Maguire, and Mark Eberhard for their assistance with this article.
This study was supported by the Ministry of Health of Brazil, CNPq PRONEX (2704) and Universidade Estadual do Norte Fluminense Darcy Ribeiro.
The institutional review board at Ministry of Health approved submission of this manuscript for publication.
(l.) Benenson MW, Takafuji ET, Lemon SM, Greenup RL, Sulzer AJ. Oocyst-transmitted toxoplasmosis associated with ingestion ingestion /in·ges·tion/ (-chun) the taking of food, drugs, etc., into the body by mouth.
1. The act of taking food and drink into the body by the mouth.
2. of contaminated water. N Engl J Med. 1982;307:666-9.
(2.) Bowie WR, King AS, Werker DH, Isaac-Renton JL, Bell A, Eng SB, et at. Outbreak of toxoplasmosis associated with municipal drinking water. The BC Toxoplasma Investigation Team. Lancet. 1997;350:173-7.
(3.) Bahia-Oliveira LMG LMG Light Machine Gun
LMG Laurence M. Gould (Antarctic Research Support Vessel, USAP)
LMG Local Marketing Group
LMG Loaf's Merry Guild
LMG Laboratory Molecular Genetics
LMG Liquid Methane Gas , Jones JL, Azevedo-Silva J, Orefice F, Crespo C, Addiss D. Highly endemic waterborne toxoplasmosis in north Rio de Janeiro Rio de Janeiro, city, Brazil
Rio de Janeiro (rē`ō də zhänā`rō, Port. rē` thĭ zhənĕē`r state, Brazil. Emerg Infect Dis. 2003;9:55-62.
(4.) Remington JS, McLeod R, Thulliez P, Desmonts G. Toxoplasmosis. In: Remington JS, Klein JO, editors. Infectious diseases of the fetus and newborn infant, 5th ed. Philadelphia: WB Saunders; 2001. p. 205-346.
(5.) Dubey JP, Ruff MD, Camargo ME, Shen Shen, in the Bible, place, perhaps close to Bethel, near which Samuel set up the stone Ebenezer. SK, Wilkins GL, Kwok OC, et al. Serologic and parasitologic responses of domestic chickens after oral inoculation with Toxoplasma gondii oocysts. Am J Vet Res. 1993;54:1668-72.
(6.) Dubey JR Refinement of pepsin pepsin, enzyme produced in the mucosal lining of the stomach that acts to degrade protein. Pepsin is one of three principal protein-degrading, or proteolytic, enzymes in the digestive system, the other two being chymotrypsin and trypsin. digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol. 1998;74:75-7.
(7.) Lehmann T, Blackstonn CR, Parmley SF, Remington JS, Dubey, JP. Strain typing of Toxoplasma gondii: comparison of antigen-coding and house keeping genes. J Parasitol. 2000;86:960-71.
(8.) da Silva A J, Bomay-Llinares F J, Moura IN, Slemenda SB, Tuttle JL, Pieniazek NJ. Fast and reliable extraction of protozoan protozoan (prō'təzō`ən), informal term for the unicellular heterotrophs of the kingdom Protista. Protozoans comprise a large, diverse assortment of microscopic or near-microscopic organisms that live as single cells or in simple parasite DNA from fecal specimens. Mol Diag. 1999;4:57-64.
(9.) Bretagne S, Costa JM, Vidaud M, Tran J, Nhieu V, Fleury-Feith J. Detection of Toxoplasma gondii by competitive DNA amplification of bronchoalveolar lavage samples. J Infect Dis. 1993; 168:1585-8.
(10.) Aramini JJ, Stephen C, Dubey JP, Engelstoft C, Schwantje H, Ribble CS. Potential contamination of drinking water with Toxoplasma gondii oocysts. Epidemiol Infect. 1999;122:305-15.
(11.) Silveira CAM. A maior Epidemia do mundo. In: Silveira CAM, editor. Toxoplasmose Duvidas e Controversias, 1st ed. Erechim: Edifapes; 2002. p. 79-82.
(12.) Howe DK, Summers BC, Sibley LD. Acute virulence in mice is associated with markers on chromosome VIII in Toxoplasma gondii. Infect Immun. 1996;64:5193-8.
(13.) Boothroyd JC, Grigg ME. Population biology of Toxoplasma gondii and its relevance to human infection: do different strains cause different disease? Curr Opin Microbiol. 2002;5:438-42.
(14.) Dubey JP, Graham DH, Seipel DS, Lehmann T, Bahia-Oliveira LMG. Toxoplasma gondii isolates of free-ranging chickens from Rio de Janeiro, Brazil: mouse mortality, genotype, and oocyst oocyst /oo·cyst/ (-sist) the encysted or encapsulated ookinete in the wall of a mosquito's stomach; also, the analogous stage in the development of any sporozoan.
n. shedding by cats. J Parasitol. 2003;89:851-3.
(15.) Dubey JP, Navarro IT, Sreekumar C, Dahl E, Freire RL, Kawabata HH, et al. Toxoplasma gondii infection in cats from Parana Brazil: seroprevalence seroprevalence Immunology The proportion of a population that is seropositive–ie, has been exposed to a particular pathogen or immunogen; the seropositivity of a population is calculated as the number of individuals who produce a particular antibody divided , tissue distribution, and biologic and genetic characterization of isolates. J Parasitol. 2004;90:721-6.
Lenildo de Moura, * (1) Lilian Maria Garcia Bahia-Oliveira, ([dagger]) (1) Marcelo Y. Wada, * Jeffrey L. Jones, ([double dagger]) Suely H. Tuboi, * Eduardo H. Carmo, * Walter Massa Massa, in the Bible
Massa (măs`ə), in the Bible, seventh son of Ishmael.
Massa, city, Italy
Massa (mäs`ä), city (1991 pop. 66,737), capital of Massa-Carrara prov. Ramalho, * Natal J. Camargo, ([section]) Ronaldo Trevisan, ([section]) Regina M. T. Graca, ([paragraph]) Alexandre J. da Silva, ([double dagger]) Iaci Moura, ([double dagger]) J.P. Dubey, (#) and Denise O. Garrett **
* Ministerio de Saude, Brasilia, Brasil; ([dagger]) Universidade Estadual do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, Brazil; ([double dagger]) Centers for Disease Control and Prevention, Atlanta, Georgia, USA; ([section]) Secretaria de Saude do Estado do Parana Curitiba, Brazil; ([paragraph]) Laboratorio Central de Saude Publica, Curitiba, Brazil; (#) United States Department of Agriculture United States Department of Agriculture (USDA),
n.pr established in 1862, USDA is responsible for the safety of meat, poultry, and egg products. It conducts ongoing research in areas from human nutrition to new crop technologies and also helps ensure open , Beltsville, Maryland; ** Centers for Disease Control and Prevention Foundation, Atlanta, Georgia, USA
Address for correspondence: Lilian Maria Garcia Bahia-Oliveira, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Centro de Biociencias e Biotecnologias, Laborat6rio de Biologia do Reconhecer. Av. Alberto Lamego 2000, Horto, Campos dos Goytacazes Campos dos Goytacazes is a municipality and city located in the northern area of Rio de Janeiro State, Brazil, with a population of 429,667 inhabitants, being the largest municipality of the state. Its area is 4.031,910 km² and its elevation is 14 m. , Rio de Janeiro, Brazil 28013-600; fax: 55-22-2726-1521 ; email: firstname.lastname@example.org
Use of trade names is for identification only and does not imply endorsement by the Public Health Service or by the U.S. Department of Health and Human Services Noun 1. Department of Health and Human Services - the United States federal department that administers all federal programs dealing with health and welfare; created in 1979
Health and Human Services, HHS .
Mr de Moura is a registered nurse and epidemiologist and member of the Training Programs in Epidemiology and Public Health Interventions Network (TEPHINET TEPHINET Training Programs in Epidemiology and Public Health Interventions Network (Atlanta, Georgia) ). His principal area of research interest is epidemiology.
Table 1. Univariate analysis showing risk factors for Toxoplasma gondii infection for statistically significant factors (positive results only), N=376 Characteristic No. persons * Case Control Water exposure Drank water exclusively from municipal tank reservoir A 350 152 198 B 26 4 22 Household tank No 95 28 67 Yes 281 128 153 Drank >10 cups water per day No 270 97 173 Yes 106 59 47 Drank beverages made with unfiltered water No 34 20 14 Yes 342 136 206 Food exposure Ate undercooked meat in past 30 days No 345 136 209 Yes 31 20 11 Ate commercial ice cream No 188 51 137 Yes 188 105 83 Ate bacon No 228 82 146 Yes 148 74 74 Ate lamb No 316 122 194 Yes 60 34 26 Ate in restaurants in the past 30 days No 277 105 172 Yes 99 51 48 95% Matched confidence Characteristic odds ratio p value interval Water exposure Drank water exclusively from 3.73 0.016 1.27-10.93 municipal tank reservoir A B Household tank 2.16 0.006 1.24-4.01 No Yes Drank >10 cups water per day 2.07 0.004 1.24-3.61 No Yes Drank beverages made with 2.25 0.044 1.02-5.50 unfiltered water No Yes Food exposure Ate undercooked meat in past 2.71 0.027 1.11-7.34 30 days No Yes Ate commercial ice cream 3.43 0.000 2.08-5.67 No Yes Ate bacon 1.89 0.009 1.15-3.02 No Yes Ate lamb 1.85 0.043 1.02-3.51 No Yes Ate in restaurants in the past 1.71 0.028 1.06-2.96 30 days No Yes * Case-patients ranged from 1 to 72 years of age (median= 28); 79 (51%) were male; 6 (3.8%) were pregnant woman. Table 2. Risk for Toxoplasma gondii infection shown as odds ratios estimated with conditional backward elimination logistic regression, N=376 Wald confidence limits Odds p Variable ratio Lower Upper value * Drinking water from reservoir A 4.55 2.01 5.49 0.001 Drinking >10 glasses of water per day 3.29 1.46 4.46 0.001 Having household water storage tank 1.81 0.99 3.33 0.054 Eating commercial ice cream 4.55 2.01 5.49 0.001 * Significant (p [less than or equal to] 0.001, rounded).