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Risk factors for Marburg hemorrhagic fever, Democratic Republic of the Congo.


We conducted two antibody surveys to assess risk factors for Marburg hemorrhagic fever Noun 1. Marburg hemorrhagic fever - a viral disease of green monkeys caused by the Marburg virus; when transmitted to humans it causes serious or fatal illness
green monkey disease, Marburg disease
 in an area of confirmed Marburg virus Marburg virus: see hemorrhagic fever.  transmission in the Democratic Republic of the Congo. Questionnaires were administered and serum samples tested for Marburg-specific antibodies by enzyme-linked immunosorbent assay enzyme-linked immunosorbent assay
n.
ELISA.


Enzyme-linked immunosorbent assay (ELISA)
A diagnostic blood test used to screen patients for AIDS or other viruses.
. Fifteen (2%) of 912 participants in a general village cross-sectional antibody survey were positive for Marburg immunoglobulin G immunoglobulin G
n. Abbr. IgG
The most abundant class of antibodies found in blood serum and lymph and active against bacteria, fungi, viruses, and foreign particles. Immunoglobulin G antibodies trigger action of the complement system.
 antibody. Thirteen (87%) of these 15 were men who worked in the local gold mines. Working as a miner (odds ratio [OR] 13.9, 95% confidence interval confidence interval,
n a statistical device used to determine the range within which an acceptable datum would fall. Confidence intervals are usually expressed in percentages, typically 95% or 99%.
 [CI] 3.1 to 62.1) and receiving injections (OR 7.4, 95% CI 1.6 to 33.2) were associated with a positive antibody result. All 103 participants in a targeted antibody survey of healthcare workers were antibody negative. Primary transmission of Marburg virus to humans likely occurred via exposure to a still unidentified reservoir in the local mines. Secondary transmission appears to be less common with Marburg virus than with Ebola virus Ebola virus (ēbō`lə), a member of a family (Filovirus) of viruses that cause hemorrhagic fevers. The virus, named for the region in Congo (Kinshasa) where it was first identified in 1976, emerged from the rain forest, where it survives in , the other known filovirus Filovirus /Fi·lo·vi·rus/ (fi´lo-vi?rus) Marburg and Ebola viruses: a genus of viruses of the family Filoviridae that cause hemorrhagic fevers (Marburg virus disease, Ebola virus disease). .

**********

Marburg hemorrhagic fever (MHF MHF Mental Health Foundation (London, UK)
MHF Malaysian Hockey Federation
MHF Method Hiding Factor
MHF Medium High Frequency
MHF Major Hazards Facility
MHF Mixed Hydrazine Fuel
MHF Magnolia House of Furniture, Inc
) is a severe illness caused by Marburg virus, a member of the Filoviridae family. MHF was first described in 1967 during outbreaks in Germany and the former Yugoslavia that were linked to monkeys imported from Uganda (1-3). Since then, only a few sporadic cases in East Africa and southern Africa
This article concerns the region in Africa. For the present-day country in this region, see South Africa; for the former country, see South African Republic.
Southern Africa
 and one laboratory infection have been identified (4-7). Serosurveys for Marburg antibodies in the general population generally have shown prevalences of <2%, indicating it to be a rare and highly lethal disease (8-25).

The largest outbreak of MHF recorded to date began in late 1998 in northeastern Democratic Republic of the Congo (DRC DRC Democratic Republic of Congo
DRC Down (Stage) Right Center
DRC Director(ate) of Reserve Components
DRC Disability Rights Commission (United Kingdom) 
) (26,27). Although the remoteness of the area and the civil war in eastern DRC delayed access and evaluation, in May 1999 a team of international investigators identified 73 cases (8 laboratory-confirmed and 65 suspected cases retrospectively identified) (28). Follow-up surveillance subsequently identified >150 cases through December 2000.

The natural reservoir Natural reservoir or nidus, refers to the long-term host of the pathogen of an infectious disease. It is often the case that hosts do not get the disease carried by the pathogen or it is asymptomatic and non-lethal.  for Marburg virus remains unknown, although it is presumed to be of zoonotic Zoonotic
A disease which can be spread from animals to humans.

Mentioned in: Zoonosis
 origin. Primary transmission of the virus from the natural reservoir appears to occur only in sub-Saharan Africa and is sometimes followed by secondary person-to-person transmission in both community and nosocomial nosocomial /noso·co·mi·al/ (nos?o-ko´me-il) pertaining to or originating in a hospital.

nos·o·co·mi·al
adj.
1. Of or relating to a hospital.

2.
 settings (4-6,29). Because of the disease's rarity and lethality, risk factors for transmission of Marburg virus have not been extensively investigated. We therefore performed two antibody surveys in the wake of the 1998-99 outbreak in DRC to explore risk factors for Marburg virus exposure and transmission. One antibody survey was a cross-sectional study cross-sectional study
n.
See synchronic study.


cross-sectional study,
n the scientific method for the analysis of data gathered from two or more samples at one point in time.
 of the general village populations; the other was a focused investigation of healthcare workers (HCWs).

Methods

Area of Study

The studies we describe were performed as an adjunct to the investigation of an outbreak of MHF in May 1999. The epicenter of the outbreak was the village of Durba in the Haut-Uele District, Oriental Province, in northeastern DRC, an isolated region approximately 200 km from the borders of Uganda and Sudan (Figure). Although no official population count for Durba is available, unofficial estimates are approximately 25,000. Watsa, a larger town of approximately 60,000 and the administrative seat of the zone, lies 14 km away. Although the Yogo ethnic group predominates, the population of Durba/Watsa is quite heterogeneous, as many people have migrated to the area to work in the local gold mines. Most are Catholic. The area has had intermittent armed conflict since the beginning of the Congolese civil war in 1996, a situation that has severely limited travel and economic growth.

[FIGURE OMITTED]

The livelihood of most of the population in the Durba/Watsa area is associated with gold mining, conducted almost exclusively by young men and most often without professional training or equipment. Some older men, women, and children are involved in the extraction of gold from ore and its sale. Subsistence farming subsistence farming

Form of farming in which nearly all the crops or livestock raised are used to maintain the farmer and his family, leaving little surplus for sale or trade. Preindustrial agricultural peoples throughout the world practiced subsistence farming.
 and hunting are also common. Although various mines exist in the area, most mining appears to take place in the Goroumbwa mine a few kilometers from the village of Durba. In addition, some miners dive in local rivers in search of gold. The existence of a hemorrhagic Hemorrhagic
A condition resulting in massive, difficult-to-control bleeding.

Mentioned in: Hantavirus Infections


hemorrhagic

pertaining to or characterized by hemorrhage.
 illness in the region appeared to be common knowledge and was labeled "Durba hemorrhagic syndrome" or "Durba syndrome" by the villagers, who often associated it with working in the mines.

Because of the remoteness of the region and the war, supplies are severely limited in all the health facilities in the region. The major facility in Durba is a small rudimentary government health center staffed by a few nurses. In Watsa, there is a larger government hospital, a hospital affiliated with the mining company, and two government health centers. In addition, at least 14 small private health centers operate collectively in Durba and Watsa.

Study Population

Two surveys were undertaken. The first was a cross-sectional survey on a convenience sample of the general population of Durba township. It was performed by establishing a post in the center of the village. With the aid of local HCWs and a village loud-speaker system, residents of sequential "quartiers" of the village were requested to come for evaluation over a 3-day period. Persons <15 years of age were excluded. The second survey focused on HCWs at all health centers in Durba and Watsa. All HCWs were surveyed at their place of employment.

Questionnaire

The rationale for conducting the study was explained to all participants, and verbal consent was obtained. Questionnaires were pretested on local villagers not included in the final study. For the general population, a 2-page questionnaire was administered. Supervisors at the local mining company were consulted about appropriate questions regarding exposures in the mines. Persons were first evaluated by local village HCWs, who determined their ability to speak and understand French. If deemed able to do so, the person was then interviewed by either a local HCW HCW Health care worker, see there  or a French-speaking member of the international investigative team. For those persons who did not understand French, the questionnaire was administered by a local HCW in the appropriate local language. Participants were asked about their entry into and activities within the mines, exposures to persons presumably pre·sum·a·ble  
adj.
That can be presumed or taken for granted; reasonable as a supposition: presumable causes of the disaster.
 sick with Durba syndrome (defined as a severe illness with high fever and bleeding from the nose, mouth, and/or anus) in the hospital and at home, and exposures to various animals thought to possibly transmit Marburg virus (rodents, bats, and monkeys). Participants were given a small bag of peanuts as a token of appreciation for their cooperation.

For the study of HCWs, a 1-page questionnaire was administered; HCWs were asked about exposure to persons with suspected Durba syndrome at work and at home, as well as any history of a compatible illness in the HCWs themselves. HCWs were also asked if they had ever entered the mines. Interviews were conducted in French.

Phlebotomy Phlebotomy Definition

Phlebotomy is the act of drawing or removing blood from the circulatory system through a cut (incision) or puncture in order to obtain a sample for analysis and diagnosis.
 and Serologic Testing

After administration of the questionnaire, 5 mL of blood was obtained and stored out of the sunlight. At the end of the day, the serum and clot were separated, labeled, and stored in liquid nitrogen Noun 1. liquid nitrogen - nitrogen in a liquid state
atomic number 7, N, nitrogen - a common nonmetallic element that is normally a colorless odorless tasteless inert diatomic gas; constitutes 78 percent of the atmosphere by volume; a constituent of all living
. Aliquots were sent on dry ice for analysis at both 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.  (CDC See Control Data, century date change and Back Orifice.

CDC - Control Data Corporation
) in Atlanta, Georgia, USA, and the National Institute for Communicable Diseases communicable diseases, illnesses caused by microorganisms and transmitted from an infected person or animal to another person or animal. Some diseases are passed on by direct or indirect contact with infected persons or with their excretions.  in Johannesburg, South Africa South Africa, Afrikaans Suid-Afrika, officially Republic of South Africa, republic (2005 est. pop. 44,344,000), 471,442 sq mi (1,221,037 sq km), S Africa. .

Testing for Marburg-specific immunoglobulin (Ig) M and IgG antibodies by the enzyme-linked immunosorbent assay (ELISA ELISA (e-li´sah) Enzyme-Linked Immuno-Sorbent Assay; any enzyme immunoassay using an enzyme-labeled immunoreactant and an immunosorbent.

ELISA
n.
) was conducted with a technique analogous to that reported for detecting antibody to Ebola virus, except for the substitution of an antigen made from the Musoke strain of Marburg virus (30,31). The cut-off value for a positive ELISA result was set to 3 standard deviations from the mean control-adjusted optical density (OD) of 410 nm found on a panel of normal serum normal serum
n.
A nonimmune serum, especially serum from an individual prior to immunization.
 samples. This value generally corresponds to an OD of approximately 0.1 at a dilution of 1:100 for the IgM assay, and 0.2 at 1:400 for the IgG assay, which generally has a higher background. Positive and negative controls were included with each run and consisted of serum from African patients with and without a laboratory-confirmed history of MHF. Because the rarity of MHF has precluded rigorous field testing of the ELISA for Marburg antibody, all ELISA-positive serum samples were also examined by the immunofluorescent immunofluorescent

having the characteristic of immunofluorescence.


immunofluorescent antibody test
see fluorescence microscopy.

immunofluorescent microscopy
see fluorescence microscopy.
 antibody assay (IFA Immunofluorescent assay (IFA)
A blood test sometimes used to confirm ELISA results instead of using the Western blotting. In an IFA test, HIV antigen is mixed with a fluorescent compound and then with a sample of the patient's blood.
), as previously described (32). The cut-off titer titer /ti·ter/ (ti´ter) the quantity of a substance required to react with or to correspond to a given amount of another substance.  for a positive IFA result was 1:50. Serum samples were considered positive only if positive results were obtained by both ELISA and IFA. Study participants were directed to seek the 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.
 results (free of charge) 1 month after testing from the nurse in charge of the local health center. The nurse was provided the results along with details on how to interpret antibody status.

Data Analysis

Questionnaires were created by using EpiInfo version 6.04 (CDC, Atlanta, GA). Data were initially recorded on the questionnaires by hand, then entered into EpiInfo 6.04, and finally imported into SPSS A statistical package from SPSS, Inc., Chicago (www.spss.com) that runs on PCs, most mainframes and minis and is used extensively in marketing research. It provides over 50 statistical processes, including regression analysis, correlation and analysis of variance.  version 10.0 statistical software (SPSS, Chicago, IL) for further analysis. The chi-square test chi-square test: see statistics. , Fisher exact test, and Student t test with Levene's test In statistics, Levene's test is an inferential statistic used to assess the equality of variance in different samples. Some common statistical procedures assume that variances of the populations from which different samples are drawn are equal.  for equality of variances were employed, where appropriate. Data found to be non-normally distributed were normalized by a log l0 transformation before statistical analysis. Statistical tests were two-sided, and significance was set at p [less than or equal to] 0.05. Variables with significance values of p [less than or equal to] 0.1 in the univariate analysis were examined in a multivariate model using forward stepwise stepwise

incremental; additional information is added at each step.


stepwise multiple regression
used when a large number of possible explanatory variables are available and there is difficulty interpreting the partial regression
 maximum likelihood logistic regression In statistics, logistic regression is a regression model for binomially distributed response/dependent variables. It is useful for modeling the probability of an event occurring as a function of other factors. .

Results

General Population

A total of 912 participants were surveyed. Another seven persons were initially enrolled but did not stay to complete their questionnaires or to have a blood sample drawn. No further information is available regarding these seven or their reasons for withdrawal. Demographic data are presented in Table 1. Since the gold deposits in Durba attract young male miners from the surrounding region, most participants were men (65%), and 347 (38%) listed their occupation as miner, although only 281 (81%) of these 347 were currently working in the mines. Virtually all (99%) of the miners were male. Marburg-specific IgG antibodies were found in 15 (2%) of the 912 study participants. All were IgM antibody negative. Thirteen (87%) of the 15 IgG antibody-positive participants were male miners. The other two were women who had never entered into the mines and whose profession was listed as "other" (most likely housewives).

All 13 of the antibody-positive miners were currently working: 10 (77%) at Goroumbwa, 2 (15%) at another mine in Durba, and 1 (8%) at an unspecified site. None reported river-diving for gold. Compared with those who were antibody-negative, antibody-positive miners tended to have worked fewer years at their present mine site but to have spent more time in the mines, although the differences were not statistically significant (Table 2). Antibody-positive miners were significantly younger than their antibody-negative counterparts (Table 1). This finding perhaps reflects the longer exposure time in the mines of younger miners (age <30 years) relative to older miners (mean [+ or -] standard error of the mean [SEM] consecutive hours per week 17.7 [+ or -] 1.2 vs. 13.7 [+ or -] 1.3, respectively, p = 0.006; mean [+ or -] SEM longest stint: 34.7 [+ or -] 2.3 vs. 23.5 [+ or -] 2.1, respectively, p < 0.001).

We examined associations between antibody to Marburg virus and various practices while working in the mines, as well as exposures related to sick persons in the home, healthcare services, and animals. In a univariate analysis, significant positive associations were found with having touched the corpse of someone who died from Durba syndrome, having had Durba syndrome oneself, and having received injections in the past year (Table 3). Touching the blood, feces, or urine of someone with Durba syndrome was of borderline statistical significance. Consumption of rodents was associated with a borderline significant protective effect, which was probably a spurious association. Of the four antibody-positive survey participants who said they had had Durba syndrome themselves, three dated the illness to the 5 months immediately before the study. The fourth, although uncertain, dated his illness to September 1998, 9 months before the investigation.

Receiving an injection as part of medical treatment was common in Durba: 505 (56%) of 907 of the participants in our cross-sectional village survey reported receiving an injection in the previous year, including 13 (87%) of the 15 antibody-positive participants (11 miners and both female nonminers). Overall, however, receiving injections was significantly more common among nonminers than miners (62 [368/596] vs. 47 [162/348], respectively, OR 1.9, 95% CI 1.4 to 2.4, p < 0.001) and among women than men (71% [221/312] vs. 49% [305/626], respectively, OR 2.6, 95% CI 1.9 to 3.4, p < 0.001).

In a multivariate model, the only variables that remained significantly associated with a positive Marburg antibody result were being a miner (OR 13.8, 95% CI 3.1 to 62.1) and having received injections (OR 7.4, 95% CI 1.6 to 33.2). Having previously had Durba syndrome was not added to the model, as it was not an independent risk factor for acquiring MHF. The associations between Marburg antibody, mining, and receiving injections remained essentially unaltered when men were looked at independently. The number of antibody-positive women (two) was too small to permit meaningful statistical analysis. However, both positive women were among the relatively few survey participants with extensive secondary contact in the household. Both reported having someone in the household sick with Durba syndrome, having contact with their body fluids, and participating in their burial, although only one of the two women reported direct contact with the corpse. In contrast, 4 (31%) of the 13 antibody-positive male miners reported any type of household exposure.

Healthcare Workers

One hundred three HCWs were enrolled from 15 different health centers, including 73 (71%) nurses, 13 (13%) clerical or administrative stall, 10 (10%) midwives, 5 (5%) laboratory workers, and 2 (2%) doctors. These figures are thought to represent virtually all of the active HCWs in the two villages except those practicing traditional medicine. HCWs had a mean of 9 years (range 0-42) of experience. All were negative for both Marburg IgM and IgG antibodies, despite the fact that 67 (65%) reported caring for a patient with Durba syndrome, and 5 (5%) reported having had Durba syndrome themselves. Types of patient contact included administering injections (38%); cleaning up blood, vomitus vomitus /vom·i·tus/ (vom´i-tus) [L.]
1. vomiting.

2. matter vomited.


vom·i·tus
n.
Vomited matter.



vomitus

1. vomiting.

2. vomited material.
, urine, or feces (28%); washing bed clothes (7%); washing corpses (6%); and receiving a needlestick injury needlestick injury Infection control The unintentional exposure of a health care worker to a needle used in direct Pt management. See Hospital-acquired penetration contacts, Sharps.  (2%).

Discussion

Despite conclusive evidence CONCLUSIVE EVIDENCE. That which cannot be contradicted by any other evidence,; for example, a record, unless impeached for fraud, is conclusive evidence between the parties. 3 Bouv. Inst. n. 3061-62.  of circulation of Marburg virus in the Durba/Watsa area in the months and years preceding our antibody surveys, we found few persons with serologic evidence of previous infection (26,27). This likely reflects a combination of the rarity of MHF and the high case-fatality ratio case-fatality ratio Epidemiology A value calculated as 100 cases of a disease 'X', divided by the number of persons with the disease who died in a given period of time; the resulting ratio is equal to the rate of a disease's occurrence. See Cause-fatality ratio.  (83%) associated with the disease in Durba/Watsa, leaving few survivors for study.

Most previous observations on risk factors for MHF have been of an anecdotal nature. Despite the small number of antibody-positive survey participants found in Durba, we were able to systematically identify and quantify several risk factors for MHF. The preponderance of antibody in male miners without obvious evidence for person-to-person transmission suggests that the local mines are a site of primary infection with Marburg virus, most likely through exposure to the primary zoonotic reservoir. Various previous findings support the conclusion of an association between MHF and exposure in mines and caves, including the following: 1) most cases of MHF identified in Durba/Watsa through December 2000 occurred in miners (J.J. Muyembe-Tamfum et al., unpub. data); 2) molecular epidemiologic data demonstrate the circulation of numerous distinct genotypes of Marburg virus in Durba/Watsa, consistent with multiple parallel primary introductions rather than a single one amplified by secondary spread (R. Swanepoel et al., unpub. data); and 3) previous cases of MHF have been associated with entry into caves (5,6).

As expected, close contact with case-patients with MHF or corpses were risk factors for secondary transmission of Marburg virus. Although injection with contaminated contaminated,
v 1. made radioactive by the addition of small quantities of radioactive material.
2. made contaminated by adding infective or radiographic materials.
3. an infective surface or object.
 syringes has been previously shown to be associated with filovirus transmission, the retrospective nature of our study makes it impossible to discern whether the use of Marburg virus-contaminated syringes resulted in virus transmission in Durba/Watsa or whether patients sick with MHF, usually a severe disease, were simply more likely to seek and receive medical care, including injections (33,34). That the general profile of the antibody-positive persons who received injections (male miners) contrasted with that of the general population (female, nonminer) suggests that the latter explanation may be more likely.

Although at least seven HCW infections have been confirmed in Durba/Watsa since 1998, we found no antibody-positive HCWs, despite what would appear to be frequent high-risk exposures (35, J.J. Muyembe-Tamfum et al., unpub. data). The high case-fatality ratio may again explain the absence of antibody-positive HCWs, although historical review does not suggest the existence of previous large nosocomial epidemics in Durba/Watsa (D. Bausch et al., unpub. data). Sound barrier nursing practices on behalf of local HCWs may have helped prevent nosocomial transmission but, given the severely limited availability When customers of the PSTN make telephone calls, they commonly make use of a telecommunications network called a switched-circuit network. In a switched-circuit network, devices known as switches are used to connect the caller to the callee.  of protective material in the area, this is unlikely to be the sole explanation.

The low prevalence of Marburg antibody found in Durba/Watsa, despite what would be considered significant risk factors for person-to-person transmission, suggests that secondary transmission of Marburg virus may be relatively infrequent compared with transmission of the other known member of the filovirus family, Ebola virus. In contrast to Ebola hemorrhagic fever Noun 1. Ebola hemorrhagic fever - a severe and often fatal disease in humans and nonhuman primates (monkeys and chimpanzees) caused by the Ebola virus; characterized by high fever and severe internal bleeding; can be spread from person to person; is largely limited to  (33,34), no large nosocomially amplified outbreaks of MHF have been noted. Only six secondary infections (five nosocomial and one sexually transmitted) were noted of the 32 cases reported during the original MHF outbreak in 1967 in Europe, despite the fact that the etiologic agent was unknown at the time of the outbreak and thus appropriate barrier nursing measures were unlikely to have been rapidly implemented (1-3,36-38). Smith et al. reported that 1 of 207 close contacts of a case-patient with MHF contracted the virus (5). Neither MHF nor antibody developed in a nurse in Durba who suffered a needlestick injury while caring for a case-patient with laboratory-confirmed and subsequently fatal MHF during the 1999 outbreak in DRC; however, the needle and IV line may have been flushed before the accident. Finally, immunohistochemical studies of skin biopsy Skin Biopsy Definition

A skin biopsy is a procedure in which a small piece of living skin is removed from the body for examination, usually under a microscope, to establish a precise diagnosis.
 specimens from patients with fatal MHF generally show that Marburg virus antigen is more sparsely distributed relative to Ebola antigen in fatal cases of Ebola hemorrhagic fever, which suggests that there may be less cutaneous cutaneous /cu·ta·ne·ous/ (ku-ta´ne-us) pertaining to the skin.

cu·ta·ne·ous
adj.
Of, relating to, or affecting the skin.


Cutaneous
Pertaining to the skin.
 shedding of Marburg virus and thus lower person-to-person communicability communicability

transmissibility; ability to spread from infected to susceptible hosts.


communicability period
the time during which the patient is infectious to others.
 in MHF (S. Zaki et al., pers. commun.).

Our study had several limitations. As participants were not randomly selected, disproportionate participation from specific subpopulations could have skewed skewed

curve of a usually unimodal distribution with one tail drawn out more than the other and the median will lie above or below the mean.

skewed Epidemiology adjective Referring to an asymmetrical distribution of a population or of data
 our results. Selective participation could have occurred because of fear of stigmatization stigmatization /stig·ma·ti·za·tion/ (stig?mah-ti-za´shun)
1. the developing of or being identified as possessing one or more stigmata.

2. the act or process of negatively labelling or characterizing another.
 or selective migrations of persons into or out of Durba/Watsa. Social stigmas could have also resulted in recall bias. The small number of Marburg antibody--positive participants limits our statistical power to identify all possible risk factors for MHF. Although ELISA testing for Marburg antibody has not undergone rigorous field testing, we believe that our conservative criterion of positive results on both ELISA and IFA for a participant to be considered Marburg antibody--positive lends credence to our conclusions. The precise duration of antibody persistence after Marburg infection is unknown for both tests. If reversion to antibody-negative status appears after a relatively short time, some previously infected persons may have escaped detection. However, most of the aforementioned limitations would likely result in false-negative results, with the ultimate effect of underestimating the magnitude of any recognized associations.

In defining risk factors for primary transmission of Marburg virus in Durba/Watsa, our study helps orient the hunt for the reservoir for the filoviruses. If primary infection to humans is indeed occurring in the mines around Durba/Watsa, future investigations of the reservoir for Marburg virus should focus on fauna present in such habitats. Bats, rodents, arthropods, and plant life within cave/mine habitats would be the prime suspects. Samples taken from small mammals captured in and around mines in Durba are being analyzed for possible Marburg virus infection (R. Swanepoel, pers. commun.). Only a combination of the use of epidemiologic and epizootiologic investigations along with direct observations made during outbreaks is likely to shed light on the still-cryptic natural history of the filoviruses.
Table 1. Demographic characteristics and Marburg immunoglobulin
G antibody results of the study population in Durba,
Democratic Republic of the Congo, 1999 (a)

                                         IgG antibody   IgG antibody
                      All participants     positive      negative
                        n = 912 (%)       n = 15 (%)    n = 897 (%)

Characteris               594 (65)         13 (87)        581 (65)
Mean age, y (range)      31 (14-79)       27 (21-42)     31 (14-79)
Profession
  Miner                   347 (38)         13 (87)        334 (37)
  Merchant                141 (15)          0 (-)         141 (16)
  Other/unknown           424 (46)          2 (13)        422 (47)

                         OR (95% (CI)       p value

Characteris           3.5 (0.8 to 15.4)       0.10
Mean age, y (range)           --              0.04
Profession                    --
  Miner               11.0 (2.5 to 48.9)     <0.001
  Merchant                    --              0.15
  Other/unknown               --               --

(a) Odds ratios (OR) and p values are for the comparison
between antibody-positive and-negative participants.

CI, confidence interval: Ig, immunoglobulin.

Table 2. Duration of time spent working in mines and Marburg
immunoglobulin G antibody status among 281 active miners in
Durba, Democratic Republic of the Congo, 1999

Time in mines              Antibody positive   Antibody negative    p
                            ([+ or -] SEM)      ([+ or -] SEM)     value
                              (n = 13)             (n = 268)

At present mine site (y)    6.6 [+ or -] 1.0   10.3 [+ or -] 0.6   0.52
Usual h/wk working in
 mine                      58.2 [+ or -] 9.2   49.5 [+ or -] 1.7   0.36
Usual h in mine without
 exiting                   24.2 [+ or -] 6.1   16.0 [+ or -] 0.9   0.17
Longest stint in mine (h)  38.8 [+ or -] 10.2  28.8 [+ or -] 1.8   0.16

Table 3. Antibody to Marburg virus and possible risk factors
for Marburg hemorrhagic fever in Durba, Democratic Republic
of the Congo, 1999 (a)

                                    All
                                participants      Antibody
Characteristic                      (%)         positive (%)

Behavior in the mines (b)
  Wear mask                       4/289 (1)    1/13 (8)
  Drink water from
    sources in the mine         160/289 (55)   9/13 (69)
  Use explosives                129/289 (45)   7/13 (54)
  Wear boots                     46/289 (16)  21/13 (15)
Household/village
    exposures to someone
    with Durba syndrome (c)
  Touched corpse                 88/905 (10)   4/15 (27)
  Touched blood,
    feces, or urine              60/903 (7)    3/15 (20)
  Worked with someone
    with syndrome               248/906 (27)  71/15 (47)
  Been in the same room with
    someone with syndrome       179/902 (20)   4/15 (27)
  Touched skin of person
    during illness              286/903 (32)   6/15 (40)
  Someone in the household
    sick with syndrome          210/906 (23)   4/15 (27)
  Participated in burial        393/904 (43)   6/15 (40)
Healthcare-related exposures
  Had Durba syndrome yourself    60/912 (7)    4/15 (27)
  Received injections
    in the last year            505/907 (56)  13/15 (87)
  Underwent surgery
    in the last year             85/905 (9)    2/15 (13)
  Received scarification (d)
    in the last year            209/906 (23)   4/15 (27)
Animal exposures
  Rodents
    Touched                     437/897 (49)   4/14 (29)
    Ate                         271/892 (30)   1/15 (7)
    Bitten by                   200/896 (22)   3/15 (20)
  Bats
    Touched                     169/901 (19)   4/14 (29)
    Ate                          31/898 (3)    0/15 (-)
    Bitten by                     8/896 (1)    0/15 (-)
  Monkeys
    Touched                     502/892 (56)   6/14 (43)
    Ate                         682/895 (76)  11/14 (79)
    Bitten by                    76/895 (8)    1/15 (7)

                                  Antibody                         p
Characteristic                  negative (%)     OR (95% CI)     value

Behavior in the mines (b)
  Wear mask                       3/276 (l)   7.6 (0. to 78.4)   0.17
  Drink water from
    sources in the mine         151/276 (55)  1.9 (0.6 to 6.2)   0.40
  Use explosives                122/276 (44)  1.5 (0.5 to 4.5)   0.57
  Wear boots                     44/276 (16)  1.0 (0.2 to 4.5)   1.00
Household/village
    exposures to someone
    with Durba syndrome (c)
  Touched corpse                 84/990 (9)   3.5 (1.1 to 11.2)  0.05
  Touched blood,
    feces, or urine              57/888 (6)   3.6 (1.0 to 13.3)  0.07
  Worked with someone
    with syndrome               241/891 (27)  2.4 (0.8 to 6.6)   0.15
  Been in the same room with
    someone with syndrome       175/887 (20)  1.5 (0.5 to 4.7)   0.51
  Touched skin of person
    during illness              280/888 (32)  1.4 (0.5 to 4.1)   0.58
  Someone in the household
    sick with syndrome          206/891 (23)  1.2 (0.4 to 3.8)   0.76
  Participated in burial        387/889 (44)  0.9 (0.3 to 2.5)   1.00
Healthcare-related exposures
  Had Durba syndrome yourself    56/997 (6)   5.4 (1.7 to 17.7)  0.01
  Received injections
    in the last year            492/892 (55)  5.2 (1.2 to 23.6)  0.02
  Underwent surgery
    in the last year             83/890 (9)   1.5 (0.3 to 6.7)   0.64
  Received scarification (d)
    in the last year            205/891 (23)  1.2 (0.4 to 3.9)   0.76
Animal exposures
  Rodents
    Touched                     433/883 (49)  0.4 (0.1 to 1.3)   0.18
    Ate                         270/877 (31)  0.2 (0.0 to 1.2)   0.05
    Bitten by                   197/881 (22)  0.9 (02 to 3.1)    1.00
  Bats
    Touched                     165/887 (19)  1.8 (0.5 to 5.6)   0.31
    Ate                          31/883 (4)          --          1.00
    Bitten by                     8/881 (1)          --          1.00
  Monkeys
    Touched                     496/878 (57)  0.6 (0.2 to 1.7)   0.42
    Ate (c)                     671/881 (76)  1.1 (0.3 to 4.2)   1.00
    Bitten by                    75/880 (9)   0.8 (0.1 to 5.9)   1.00

(a) Odds ratios (OR) and p values are for the comparison between
antibody-positive and -negative participants. CI, confidence
interval.

(b) Includes only responses from persons who stated that they
currently worked in the mines.

(c) Before questioning began, Durba syndrome was described to the
participant as "a severe illness with high fever and bleeding from
the nose, mouth, and/or anus."

(d) Scarification is the practice of intentionally scarring the
skin with sharp instruments. It may be done for aesthetic reasons
of a belief that it has medicinal or spiritual value.

(e) Many participants reported the meat was smoked or cured at
the time of purchase, so potential exposure to viable may have
been limited.


Acknowledgments

We thank the following for their assistance in this study: Ray Arthur, Angelo Belli, Leo Leo, in astronomy
Leo [Lat.,=the lion], northern constellation lying S of Ursa Major and on the ecliptic (apparent path of the sun through the heavens) between Cancer and Virgo; it is one of the constellations of the zodiac.
 Braack, Michel Noureddine Kassa, Alan Kemp, Leon Kinuani, Herwig Leirs, Laura Morgan, Jean-Pierre Mustin, Camille Nakwa, C.J. Peters, Antoine Tshomba, Florimond Tshioko, Kent Wagoner, Herve Zeller, and the administration of the Kilo-Moto (OKIMO) mining company in Durba, Democratic Republic of the Congo.

Financial support for the investigation of Marburg hemorrhagic fever in the Democratic Republic of the Congo, of which this study was a part, was received from the Office of Foreign Disaster Assistance The Office of U.S. Foreign Disaster Assistance (OFDA) is the office within USAID responsible for directing and coordinating U.S. Government relief assistance overseas. This office is within the USAID Bureau of Democracy, Conflict, and Humanitarian Assistance (DCHA).  of the United States Agency for International Development The United States Agency for International Development (or USAID) is the U.S. government organization responsible for most non-military foreign aid. An independent federal agency, it receives overall foreign policy guidance from the U.S. .

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Daniel G. Bausch, * (1) Matthias Borchert, ([dagger]) (2) Thomas Grein, ([double dagger]) Cathy Roth, ([double dagger]) Robert Swanepoel, ([section]) Modeste L. Libande, ([paragraph]) Antoine Talarmin, (#) (3) Eric Bertherat, ** (4) Jean-Jacques Muyembe-Tamfum, ([paragraph]) Ben Tugume, ([dagger]) ([dagger]) Robert Colebunders, ([dagger]) Kader M. Konde, ([double dagger] [double dagger] 5) Patricia Pirard, ([subsections]) Loku L. Olinda, ([paragraph]) Guenael R. Rodier, ([double dagger]) Patricia Campbell, ([paragraphs]) Oyewale Tomori, ([double dagger] [double dagger]) Thomas G. Ksiazek, * and Pierre E. Rollin *

* Centers for Disease Control and Prevention, Atlanta Georgia, USA; ([dagger]) Institute of Tropical Medicine tropical medicine, study, diagnosis, treatment, and prevention of certain diseases prevalent in the tropics. The warmth and humidity of the tropics and the often unsanitary conditions under which so many people in those areas live contribute to the development and , Antwerp, Belgium; ([double dagger]) World Health Organization, Geneva Geneva, canton and city, Switzerland
Geneva (jənē`və), Fr. Genève, canton (1990 pop. 373,019), 109 sq mi (282 sq km), SW Switzerland, surrounding the southwest tip of the Lake of Geneva.
, Switzerland; ([section]) National Institute for Communicable Diseases, Johannesburg, South Africa; ([paragraph]) Ministry of Health, Kinshasa, Democratic Republic of the Congo; (#) Institut Pasteur, Cayenne, French Guiana; ** Le Pharo, Marseille, France; ([dagger]) ([dagger]) Uganda Virus Research Institute The Uganda Virus Research Institute (UVRI), located in Entebbe, Uganda, was established in 1936 as the Yellow Fever Research Institute by the Rockefeller Foundation. , Entebbe, Uganda; ([double dagger]) ([double dagger]) World Health Organization, AFRO, Harare, Zimbabwe; ([subsections]) Doctors without Borders Doctors Without Borders, Fr. Médecins Sans Frontières (MSF), international organization that provides emergency medical assistance to people suffering from a natural or societal disaster, such as an earthquake or war. , Brussels, Belgium; and ([paragraphs]) Doctors without Borders, Amsterdam, the Netherlands

(1) Present address: Tulane School of Public Health and Tropical Medicine, New Orleans, LA, USA.

(2) Present address: London School of Hygiene and Tropical Medicine, London, England.

(3) Present address: Institut Pasteur, Bangui, Central African Republic.

(4) Present address: World Health Organization, Geneva, Switzerland.

(5) Present address: World Health Organization, AFRO, Ouagadougou, Burkina Faso.

Dr. Bausch is a medical epidemiologist specializing in the investigation and control of viral hemorrhagic fevers.

Address for correspondence: Daniel G. Bausch, Tulane School of Public Health and Tropical Medicine, Department of Tropical Medicine, SL-17, J. B. Johnston Building, Rm. 511, 11430 Tulane Avenue, New Orleans, LA 70112-2699 USA; fax: 504-988-6686; email: dbausch@Tulane.edu
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