Prospective study of etiologic agents of community-acquired pneumonia in patients with HIV infection.

OBJECTIVES: To study prospectively HIV-positive patients admitted to the hospital because of pneumonia by extensive laboratory tests to determine specific microbiologic diagnoses and to establish the best clinical diagnosis after review of all available data by expert clinicians. METHODS: Patients admitted to one of two hospitals had extensive questionnaires completed and defined diagnostic tests performed on blood, sputum, urine and bronchoalveolar lavage specimens, when available. RESULTS: A total of 230 patients had a diagnosis of pneumonia verified. A definite or probable etiologic diagnosis was made in 155 (67%) of these patients. Pneumocystis carinii caused 35% of all cases of pneumonia. Twenty-seven percent of cases of pneumonia with a single etiology had a definite or probable bacterial etiology. 'Atypical agents' were distinctly uncommon. Few clinical or laboratory parameters could differentiate specific etiologies. CONCLUSIONS: P. carinii continues to be a common cause of pneumonia in these patients. The rarity of 'atypical agents' could simplify the empiric approach to therapy. Despite the use of extensive testing we did not find a definite etiology in a large number of cases.

Multiyear prospective study of intestinal parasitism in a cohort of Peace Corps volunteers in Guatemala.

We conducted a prospective, longitudinal study in a cohort of 36 Peace Corps volunteers (PCVs) in Guatemala to study the incidence and natural history of intestinal parasitic infections during the PCVs' >2-year overseas stay. PCVs collected stool specimens at least monthly and when ill with gastrointestinal symptoms. Of the 1,168 specimens tested, 453 (38.8%) were positive for at least one parasite and 48 (4.1%) were positive for a pathogenic parasite. A median interval of 187 days (range, 14 to 752 days) elapsed before the first documented parasitic infection, and the median intervals from arrival until subsequent infections (e.g., second or third) were >300 days. The PCVs had 116 episodes of infection with 11 parasites, including up to 4 episodes per PCV with specific nonpathogens and Blastocystis hominis. The incidence, in episodes per 100 person-years, was highest for B. hominis (65), followed by Entamoeba coli (31), Cryptosporidium parvum (17), and Entamoeba hartmanni (17). The PCVs' B. hominis episodes lasted 6,809 person-days (28.7% of the 23,689 person-days in the study), the E. coli episodes lasted 2,055 person-days (8.7%), and each of the other types of episodes lasted <2% of the person-days in the study. Gastrointestinal symptoms were somewhat more common and more persistent, but not significantly so, in association with pathogen episodes than with B. hominis and nonpathogen episodes. Although infections with pathogenic parasites could account for only a minority of the PCVs' diarrheal episodes, the continued acquisition of parasitic infections throughout the PCVs' >2-year stay in Guatemala suggests that PCVs repeatedly had fecal exposures and thus were at risk for infections with both parasitic and nonparasitic pathogens throughout their overseas service.

Malaria, intestinal parasites, and schistosomiasis among Barawan Somali refugees resettling to the United States: a strategy to reduce morbidity and decrease the risk of imported infections.

In 1997, enhanced health assessments were performed for 390 (10%) of approximately 4,000 Barawan refugees resettling to the United States. Of the refugees who received enhanced assessments, 26 (7%) had malaria parasitemia and 128 (38%) had intestinal parasites, while only 2 (2%) had Schistosoma haematobium eggs in the urine. Mass therapy for malaria (a single oral dose of 25 mg/kg of sulfadoxine-pyrimethamine) was given to all Barawan refugees 1-2 days before resettlement. Refugees >2 years of age and nonpregnant women received a single oral dose of 600 mg albendazole for intestinal parasite therapy. If mass therapy had not been provided, upon arrival in the United States an estimated 280 (7%) refugees would have had malaria infections and 1,500 (38%) would have had intestinal parasites. We conclude that enhanced health assessments provided rapid on-site assessment of parasite prevalence and helped decrease morbidity among Barawan refugees, as well as, the risk of imported infections.

Declining CD4+ T-lymphocyte counts are associated with increased risk of enteric parasitosis and chronic diarrhea: results of a 3-year longitudinal study.

From January 1991 through September 1994, we observed people who were infected with HIV to assess the impact of enteric parasite-associated diarrhea. Respondents answered comprehensive questionnaires covering clinical and epidemiologic information and provided stool specimens monthly, which were examined unstained as well as stained with trichrome, chromotrope 2R, and with Kinyoun carbol-fuchsin, and with indirect immunofluorescence for Cryptosporidium. In all, 602 participants, who were interviewed, provided stool specimens at 3254 monthly visits. Parasites were associated with 50 of 354 (14.1%) acute diarrheal episodes (lasting < or = 28 days) and with 97 of 279 (34.8%) chronic episodes (lasting > 28 days). A parasite was associated with 31 of 222 (14.0%) episodes that occurred when CD4+ counts were > or = 200 cells/microl and with 150 of 566 (26.5%) episodes that occurred when CD4+ counts were < 200 cells/microl. The most commonly identified parasite was C. parvum, which was associated with 18 of 354 (5.1%) acute episodes and 36 (12.9%) of the 279 chronic episodes of diarrhea. In this patient population, enteric protozoan parasites were commonly associated with illness, particularly as immunosuppression worsened, and were more likely to be associated with chronic rather than acute diarrhea.

An outbreak of cyclosporiasis in Florida in 1995: a harbinger of multistate outbreaks in 1996 and 1997.

Before 1995, only one outbreak of cyclosporiasis had been reported in the United States. To identify risk factors for Cyclospora infection acquired in Florida in 1995, we conducted a matched case-control study (24 sporadic cases and 69 controls) and retrospective cohort studies of clusters of cases associated with two May social events (attack rates = 15.4% [8 of 52] and 54.5% [6 of 11]). In univariate analysis of data from the case-control study, consumption of fresh raspberries (odds ratio [OR] = 6.0, 95% confidence interval [CI] = 1.1-31.7) and bare-handed contact with soil (OR = 5.4, 95% CI = 1.4-20.7) were associated with infection; soil contact was also implicated in multivariate analysis. For the events, mixed-fruit items that had only fresh raspberries and strawberries in common had elevated relative risks (3.7 and 4.2), but the confidence intervals overlapped 1.0. The raspberries eaten at the events and by sporadic case-patients were imported. Given the cumulative evidence of the three studies and the occurrence in 1996 and 1997 of outbreaks in North America associated with consumption of Guatemalan raspberries, food-borne transmission of Cyclospora was likely in 1995 in Florida as well.

Enzyme-linked immunoelectrotransfer blot analysis of a cryptosporidiosis outbreak on a United States Coast Guard cutter.

Symptoms consistent with an outbreak of cryptosporidiosis (diarrhea, vomiting, nausea, and abdominal cramps) occurred on a U.S. Coast Guard cutter within 0-18 days after the cutter filled its tanks with Milwaukee, Wisconsin city water in March 1993. At three-weeks postdocking (PD), the suspected water was removed, and serum samples and stool specimens were collected from 47 of the 58 crew members, as well as questionnaire data on their water consumption and symptoms aboard the cutter. At 10-weeks PD and/or at 28-weeks PD, additional serum specimens were collected. Intensitometric data from enzyme-linked immunoelectrotransfer blot (EITB) were obtained on IgA responses to a 17-kD antigen group, IgM responses to a 27-kD antigen group, and IgG responses to 27-, 17-, and 15-kD antigen groups extracted from oocysts. In addition, IgG responses to crude oocyst antigens were obtained by ELISA. Based on reported symptoms, EITB results, and stool examination, the crew members were classified as confirmed (10), probable (10), suspected (22), and noncases (16). Of the 10 confirmed cases (all symptomatic) and the 10 probable cases (eight symptomatic) whose stools were positive and negative, respectively, for Cryptosporidium oocysts by microscopy, all showed changes in EITB intensities to the antigen groups and were considered EITB positive. The remaining 38 crew members, 22 suspected cases (all symptomatic), and 16 noncases (all asymptomatic), if tested, had negative stool examinations and were considered EITB negative. Of the 10 confirmed cases, only four showed a significant change in IgG responses (P < 0.05) between three-weeks PD and follow-up serum specimens by ELISA. Crew members considered confirmed cases consumed significantly more water (P < or = 0.005) aboard the cutter than noncases. Crew members considered EITB positive consumed more water (P < or = 0.04) than crew members considered EITB negative while there was no significant difference in water consumption (P > or = 0.19) between crew members considered ELISA positive and ELISA negative. Using the EITB, the observation of changes in intensity of IgA responses to the 17-kD antigen group, IgM responses to the 27-kD antigen group, and IgG responses to the 27- 17-, and 15-kD antigen groups from C. parvum oocysts between acute and convalescent serum specimens appears useful for immunodiagnosis of Cryptosporidium infection and for prospective epidemiologic studies designed to monitor infection risk.

First reported outbreak in the United States of cryptosporidiosis associated with a recreational lake.

In the summer of 1994, an outbreak of cryptosporidiosis occurred among visitors to a state park in New Jersey. We enrolled 185 persons in a cohort study, 38 (20.5%) of whom had laboratory-confirmed cryptosporidiosis or gastrointestinal illness that met our clinical case definition. Having any exposure to lake water (e.g., swimming) was strongly associated with illness (P < .001). The outbreak lasted 4 weeks and affected an estimated 2,070 persons. The most likely sources of the outbreak were contaminated runoff of rainwater and infected bathers. This outbreak of cryptosporidiosis is the first reported to be associated with recreational exposure to lake water. Our investigation shows that even a large and ongoing outbreak may not be detected for several weeks. Health professionals and persons at high risk for severe cryptosporidiosis should be aware that recreational water can be a source of cryptosporidium infection.

Etiology of bloody diarrhea in Bolivian children: implications for empiric therapy. Bolivian Dysentery Study Group.

In Bolivia, few data are available to guide empiric therapy for bloody diarrhea. A study was conducted between December 1994 and April 1995 to identify organisms causing bloody diarrhea in Bolivian children. Rectal swabs from children <5 years old with bloody diarrhea were examined for Salmonella, Shigella, and Campylobacter organisms; fecal specimens were examined for Entamoeba histolytica. A bacterial pathogen was identified in specimens from 55 patients (41%). Shigella organisms were found in 39 specimens (29%); 37 isolates (95%) were resistant to ampicillin, 35 (90%) to trimethoprim-sulfamethoxazole, and 24 (62%) to chloramphenicol, but all were susceptible to nalidixic acid. Only 1 of 133 stool specimens contained E. histolytica trophozoites. Multidrug-resistant Shigella species are a frequent cause of bloody diarrhea in Bolivian children; E. histolytica is uncommon. Clinical predictors described in this study may help identify patients most likely to have Shigella infection. Laboratory surveillance is essential to monitor antimicrobial resistance and guide empiric treatment.

Factors influencing Cryptosporidium testing in Connecticut.

To describe patterns of testing for Cryptosporidium oocysts in stool samples, Connecticut laboratories were surveyed. Different detection methods were used. Most laboratories examined stools specifically for Cryptosporidium only on physician request. The rate of positive tests varted widely (0 to 28%). Higher rates of positivity were associated with the use of monoclonal antibody methods, the use of two or more staining procedures, and testing of stool specimens in addition to those requested by physicians.

Cryptosporidial and microsporidial infections in human immunodeficiency virus-infected patients in northeastern Brazil.

To determine the frequency of the parasitic pathogens in human immunodeficiency virus (HIV)-infected patients in a developing world setting, 295 stool specimens were examined from 166 HIV-positive patients (49% with AIDS) at São José Hospital, Fortaleza, Brazil, from September 1990 to March 1992. Significantly more patients with diarrhea (85%) than without (66%) had AIDS or AIDS-related complex (ARC) (P < .005). Of the potential parasitic causes of diarrhea, only Cryptosporidium parvum and microsporidia were significantly associated with diarrheal disease. Infections with C. parvum, but not microsporidia, were associated with the rainy season (P < .005). Thus, C. parvum and microsporidia are the most common intestinal parasites associated with diarrhea in an HIV-infected population in Brazil and are associated with advanced HIV disease. The association of C. parvum infections with the rainy season suggests that contaminated water may be important in its transmission; however, the source of human microsporidia requires further investigation.

Infections with intestinal parasites in Peace Corps volunteers in Guatemala.

To assess the role of parasites in causing diarrhea in Peace Corps volunteers in Guatemala, 115 stool specimens from a case-control investigation (48 case [diarrhea] and 26 control episodes) were examined. A potentially pathogenic protozoan that could account for diarrheal illness was found for only 12% of the case episodes.

Epidemic Shigella dysenteriae type 1 in Burundi: panresistance and implications for prevention.

An epidemic of Shigella dysenteriae type 1 infections has affected Africa since 1979. Reported dysentery cases increase sharply in Burundi during September through December. Of stool samples from 189 patients reporting bloody diarrhea in November 1990, a pathogen was identified in 123 (65%). The pathogen was S. dysenteriae type 1 in 82 (67%). All S. dysenteriae type 1 isolates were resistant to ampicillin, chloramphenicol, nalidixic acid, streptomycin, sulfisoxazole, tetracycline, and trimethoprim-sulfamethoxazole. Thirty-two specimens (26%) yielded other Shigella species. Patients with S. dysenteriae type 1 were more likely than those with other Shigella infections to have abdominal pain, "lots of blood" in the stool, blood in the stool specimen examined by the interviewer, recent contact with a person with dysentery, or recent antimicrobial treatment. Thus, the seasonal increase in dysentery was due largely to multidrug-resistant S. dysenteriae type 1, clinical and epidemiologic features may predict such infection, and efforts to control this epidemic must focus on preventing transmission.

Giardia lamblia and Cryptosporidium infections in child day-care centers in Fulton County, Georgia.

Risk factors for the introduction, spread and persistence of Cryptosporidium and Giardia lamblia infections in child day-care centers are not well understood. In 1989 and 1990 stool specimens were obtained from 292 diapered children attending 17 randomly selected day-care centers in Fulton County, GA; 8 (2.7%) children in 2 centers were infected with Cryptosporidium and 21 (7.2%) children in 7 centers were infected with Giardia. In 1986 the prevalence of Cryptosporidium and Giardia in these same centers had been 0.4 and 11.0%, respectively; the prevalence of Cryptosporidium, but not Giardia, increased significantly (P = 0.04) between 1986 and 1989 to 1990. Risk factors for Giardia infection included day-care attendance for greater than 3 months, the presence of toddlers in the classroom and the presence of other children in the household. Day-care centers with a Giardia-positive child in 1986 were not more likely to have an infected child in 1989 to 1990. Cryptosporidium, like Giardia, may be endemic in day-care centers in Fulton County.

Use of pooled formalin-preserved fecal specimens to detect Giardia lamblia.

Three formalin-preserved fecal specimens from the same child attending a child-care center were pooled and compared with the three separate individual specimens by a single microscopic examination of concentration sediment for Giardia lamblia. The sensitivity of the pooled system was 100% when two or more individual specimens were positive and 88% when only one individual specimen was positive. The organism density in a single specimen was not a factor of whether the pool of specimens was positive or negative. Nearly half of the pools that contained positive specimens had only one of three specimens with positive results, reinforcing the need for multiple stool examinations when diagnosing G. lamblia infections.

Threshold of detection of Cryptosporidium oocysts in human stool specimens: evidence for low sensitivity of current diagnostic methods.

To determine the minimum number of Cryptosporidium oocysts that can be detected in stool specimens by diagnostic procedures, stool samples seeded with known numbers of Cryptosporidium parvum oocysts were processed by the modified Formalin-ethyl acetate (FEA) stool concentration method. FEA concentrates were subsequently examined by both the modified cold Kinyoun acid-fast (AF) staining and fluorescein-tagged monoclonal antibody (immunofluorescence [IF]) techniques. Oocysts were more easily detected in watery diarrheal stool specimens than they were in formed stool specimens. For watery stool specimens, a 100% detection rate was accomplished at a concentration of 10,000 oocysts per g of stool by both the AF staining and IF techniques. In formed stool specimens, 100% of specimens seeded with 50,000 oocysts per gram of stool were detected by the IF technique, whereas 500,000 oocysts per g of stool were needed for a 100% detection rate by AF staining. Counting of all oocysts on IF slides indicated a mean oocyst loss ranging from 51.2 to 99.6%, depending on the stool consistency as determined by the FEA concentration procedure. Our findings suggest that the most commonly used coprodiagnostic techniques may fail to detect cryptosporidiosis in many immunocompromised and immunocompetent individuals.

Evaluation of a commercially available enzyme-linked immunosorbent assay for Giardia lamblia antigen in stool.

The lack of a quick, simple, and inexpensive diagnostic test has limited the ability of public health officials to rapidly assess and control outbreaks of Giardia lamblia in child day-care centers. We evaluated the performance of a commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of a G. lamblia-associated antigen in stool. Stool specimens were collected from the diapers of 426 children attending 20 day-care centers, fixed in 10% Formalin and polyvinyl alcohol, and examined by microscopy by Formalin concentration and trichrome staining techniques. Specimens were also tested visually and spectrophotometrically by ELISA. Of 99 tests positive by microscopy, 93 were visually positive by ELISA (sensitivity, 93.9%). Of 534 tests negative for G. lamblia by microscopy, 32 (6.0%) were ELISA positive. However, on the basis of examination of multiple specimens from the same child, none of these could be considered false-positive ELISAs; the specificity of the ELISA was therefore 100%. The sensitivity of both microscopy and ELISA improved as the number of specimens per child increased. An optical density value of greater than 0.040 was 98.0% sensitive and 100% specific for G. lamblia. This ELISA, which appeared to be more sensitive for G. lamblia than did microscopic examination of stool, should be useful as an epidemiologic tool, particularly in day-care settings, and may also have a role in confirming clinical diagnoses of giardiasis.

Sodium azide: ineffective as a faecal preservative for parasitological diagnosis.

Sodium azide was compared with 10% formalin to evaluate sodium azide's effectiveness as a faecal preservative for intestinal helminths and protozoa. Faecal specimens collected from Haiti were preserved in sodium azide and in 10% formalin and analysed after 1.5, 6.5 and 11.5 weeks by examining direct wet-mount preparations. Sodium azide did not preserve the morphology of either helminths or protozoa as well as 10% formalin did. However, sodium azide prevented embryogenesis of helminth eggs, while some helminth eggs in 10% formalin contained living larvae. Biosafety guidelines regarding the toxicity, reactivity, and disposal of sodium azide were strictly followed. Use of 10% formalin is a significantly better choice than sodium azide for preserving parasites when accurate identification of parasites and biosafety are the main concerns.

Recommendations for collection of laboratory specimens associated with outbreaks of gastroenteritis.

Recent discoveries have implicated a number of "new" (i.e., previously unrecognized) infectious agents as important causes of outbreaks of gastroenteritis. Unfortunately, the ability to detect these agents in an outbreak can be limited by two factors: 1) the lack of appropriate assays-many of which are still in developmental stages and are not readily available to clinical laboratories, and 2) inadequately or improperly collected specimens. At CDC, many newly developed assays are being used for research and for outbreak investigations. The information in this report is especially intended for public health agencies that collaborate with CDC in investigating outbreaks of gastroenteritis. The report provides an update on guidelines and recommendations for the proper collection of specimens to be sent to CDC, gives general background information concerning some recently discovered pathogens, lists some of the tests available at CDC, and provides a list of CDC contacts. The guidelines and the general information provided on causes of outbreaks of gastroenteritis can be also used by public health workers for investigations when specific testing is available and appropriate.

Dietary fiber and giardiasis: dietary fiber reduces rate of intestinal infection by Giardia lamblia in the gerbil.

Gerbils were maintained on a low-fiber (5%) or a high-fiber (20%) diet in which the major fiber source was cellulose. Animals in the low-fiber diet group were significantly more likely to become infected when inoculated with 100 Giardia lamblia cysts than were animals in the high-fiber group. No differences were detected in gastrointestinal transit, gastric, and small intestinal luminal pH, or in duodenal mucus blanket acidic glycoprotein between animals in the high- and the low-fiber diet groups at the time of cyst inoculation. The fiber content of the diet after cyst inoculation determined the infection rate. These data suggest that the dietary fiber effect occurred during trophozoite colonization of the small intestine. When infected animals on the low-fiber diet were placed on the high-fiber diet for 24 hr, trophozoite clearing occurred in the lower small intestine. In the jejunum, the number of trophozoites attached to the mucosal surface decreased, while the number associated with luminal mucus increased. We conclude that the fiber-induced mucus secretion and the bulk movement of the insoluble fiber reduced the attachment of trophozoites to the intestinal mucosa, which decreased the probability of trophozoites establishing and sustaining colonization of the mucosa.