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Crimean-congo hemorrhagic fever, Thogoto, dugbe, and Jos viruses isolated from ixodid ticks in Ethiopia.

From 1974 through 1976, U.S. Naval Medical Research Unit No. 5 isolated 25 strain of tick-borne virus in infant mice from 410 pools containing over 6,000 ticks, and one strain from a bird and one strain from a rodent collected in central and southern Ethiopia. Of these, 17 were identified as known viruses previously found in West Central and East Africa. There were 8 strains of Jos virus from Amblyomma ticks; 7 strains of Dugbe virus from a bird, a rodent and from ticks; 1 strains of Crimean-Congo hemorrhagic fever virus and 1 strain of Thogoto virus from ticks.

Animals↗

Close relationship of Crimean hemorrhagic fever-Congo (CHF-C) virus strains by neutralizing antibody assays.

Strains of Crimean hemorrhagic fever-Congo (CHF-C) virus originating in widely separated points representing diverse zoogeograhiczl and ecological areas were compared by a neutralization of fluorescent foci of infection-test, in an attempt to determine whether antigenic differences could be detected among the strains. Minor differences were found which could be confirmed by reciprocal kinetic neutralization tests with one-injection immune sera if the results were analyzed by linear regression. However, CHF-C virus strains cannot be reliably subdivided into geographic variants on the basis of the minor antigenic variations observed. In this apparent paucity of antigenic diversity over its wide distribution, CHF-C virus differs from other arboviruses with similar or even lesser distribution.

Animals↗

Immunosorbent assays for diagnosis of Crimean-Congo hemorrhagic fever (CCHF).

Utilization of direct and indirect enzyme-linked immunosorbent assay (ELISA) and solid-phase radioimmunoassay (SPRIA) for the diagnosis of Crimean-Congo hemorrhagic fever (CCHF) allows the detection of low amounts of infectious virus (2 log LD50) or inactivated antigen and antibody to CCHF within 5-6 hours. These methods were shown to be more sensitive, specific, rapid, and reproducible than the complement-fixation test, immunofluorescence, hemagglutination, or radial diffusion in gel. The experimental design of ELISA and SPRIA developed for CCHF may be used successfully for the detection of the other members of the Bunyaviridae family.

Animals↗

Crimean-congo hemorrhagic fever in South Africa.

Crimean-Congo hemorrhagic fever virus was isolated for the first time in South Africa in February 1981, from the blood of a 13-year-old boy who died in Johannesburg after attending a camp in a nature reserve in the western Transvaal. Virus was isolated from 21/120 pools of questing ticks from the nature reserve, the infected species being Hyalomma marginatum rufipes and H. truncatum. Virus was also isolated from 4/38 pools of partially engorged ticks and other ectoparasites collected off hosts, the infected species being H.m. rufipes, H. truncatum and Rhipicephalus evertsi. Antibodies were found in the sera of 5/74 humans, 8/26 wild vertebrates, 74/270 sheep, and 109/170 cattle from the reserve and surrounding farms. Antibodies were also found in 28/200 hares from various locations in the country. It was concluded that the virus is widely prevalent in South Africa, but the full medical and veterinary significance of its presence has yet to be determined.

Adolescent↗

Reversed passive hemagglutination and inhibition with Rift Valley fever and Crimean-Congo hemorrhagic fever viruses.

Reversed passive hemagglutination (RPHA) tests were performed by coating glutaraldehyde-fixed and tannic acid-treated sheep erthrocytes with antibodies to Rift Valley fever (RVF) and Crimean-Congo hemorrhagic fever (CCHF) viruses. Cells coated with crude antibody, in the form of mouse immune ascitic fluid, reacted with high specificity and sensitivity in RPHA tests with live or inactivated virus antigens. In inhibition (RPHI) tests, RVF antibody titers in human and sheep sera were similar to those determined by hemagglutination inhibition test. RVF virus antigen could be detected in viremic sheep sera by the RPHA technique. RPHI antibody titers to CCHF virus in human and hare sera were similar to indirect immunofluorescence (IF) titers, but sheep and cattle sera with RPHI titers of 1:32 or less were negative in IF tests.

Animals↗

Epidemiologic and clinical features of Crimean-Congo hemorrhagic fever in southern Africa.

Following the diagnosis in 1981 of the first case of Crimean-Congo hemorrhagic fever (CCHF) in South Africa, an antibody survey was undertaken on cattle sera to determine the distribution of the virus and specific diagnostic tests were routinely applied to specimens from suspected cases of hemorrhagic fever to establish the medical significance of its presence. Antibody to CCHF virus was demonstrated by reversed passive hemagglutination-inhibition technique in 2,460/8,667 (28%) cattle sera and in 140/180 herds tested in South Africa, as well as in 347/763 (45%) cattle sera and in 32/34 (94%) herds tested in Zimbabwe. The antibody was found in all major cattle farming areas, but was of low prevalence along the southern coast where 2 of the 3 species of Hyalomma tick which occur in South Africa are absent. From February 1981 to January 1986, inclusive, 29 indigenous cases of CCHF were diagnosed in 16 outbreaks which arose in various locations throughout South Africa. A further 2 imported cases of CCHF arose in Zaire and Tanzania. The clinical features of infection conformed to the classical descriptions of CCHF in the Soviet Union. The fatal outcome in 11/31 cases indicates that the African disease is no less severe than that which occurs in Eurasia. It is inferred that the virus is widespread in all countries in Africa and Eurasia which lie within the limits of world distribution of ticks of the genus Hyalomma.

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Antibody to Crimean-Congo hemorrhagic fever virus in wild mammals from southern Africa.

Crimean-Congo hemorrhagic fever (CCHF) virus is becoming increasingly recognized as an important human pathogen in southern Africa. In order to determine the role of wild mammals in the natural ecology of the virus, sera from 3,772 wild mammals of 87 species and from 1,978 domestic dogs collected in South Africa and Zimbabwe between 1964 and 1985 were tested for antibody to CCHF virus by reversed passive hemagglutination inhibition (RPHI) and by indirect immunofluorescence (IF). Antibody was found to be highly prevalent in large mammals in the Orders Artiodactyla and Perissodactyla such as giraffe, Giraffa camelopardalis (3/3 positive), rhinoceros, Ceratotherium simium and Diceros bicornis (7/13), eland, Taurotragus oryx (59/127), buffalo, Syncerus caffer (56/287), kudu, Tragelaphus strepsiceros (17/78), and zebra, Equus burchelli (16/93). In small mammals antibody was found in the sera of 40/293 hares, 22/1,305 rodents, and 1/74 wild carnivores, but not in 522 primates, 176 insectivores, or 19 hyrax. Antibody was also found in the sera of 118/1,978 domestic dogs. The species of wild mammal in which antibody was distributed (with highest antibody prevalence in hares and large herbivores) reflects the feeding preference of immature and adult ticks of the genus Hyalomma, suggesting that Hyalomma sp. are the principal CCHF vectors in the wild.

Animals↗

Preparation and use of monoclonal antibodies for identifying Crimean-Congo hemorrhagic fever virus.

Seven monoclonal antibodies were prepared against a South African strain of Crimean-Congo hemorrhagic fever (CCHF) virus and were found to be directed against viral nucleocapsid protein. Five of the monoclonal antibodies reacted to high titer in indirect immunofluorescence (IF) tests and enzyme-linked immunosorbent assays (ELISA) with 22 strains of CCHF virus and failed to cross-react with the closest antigenic relative of CCHF, Hazara virus, or with 4 other nairoviruses which need to be distinguished from CCHF virus in Africa. These antibodies, used in the IF technique, readily detected antigens induced by all strains of CCHF virus included in the study in cell culture monolayers and mouse brain tissue, which represent the systems commonly used for isolation of CCHF virus. The IF technique with monoclonal antibodies constitutes a rapid and specific means of identifying newly isolated strains of CCHF virus.

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Experimental transmission of Crimean-Congo hemorrhagic fever virus by Hyalomma truncatum Koch.

Larval Hyalomma truncatum ticks were infected with Crimean-Congo hemorrhagic fever (CCHF) virus by allowing them to engorge on viremic newborn mice. The overall tick infection rate was 4.4% (24/542). Virus was detected in specimens for greater than or equal to 160 days postinfection. Transstadial transmission to the adult tick stage was observed and horizontal transmission to a mammalian host was demonstrated. Horizontal transmission of CCHF virus to uninfected adult ticks occurred while feeding with transstadially infected ticks on the same host. No evidence of transovarial virus transmission from infected female ticks to their 1st generation progeny was observed.

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Experimental studies on the replication and transmission of Crimean-Congo hemorrhagic fever virus in some African tick species.

Seven African tick species were studied as potential vectors of Crimean-Congo hemorrhagic fever (CCHF) virus. Engorged nymphae of 4 ixodid species, Hyalomma marginatum rufipes, H. truncatum, Rhipicephalus evertsi mimeticus, and Amblyomma hebraeum, were inoculated intracoelomically with CCHF virus and assayed for virus content at varying times post-inoculation. The virus replicated in all 4 species, reaching maximum titers of 4.6-5.5(10) fluorescence focus units per ml on days 5-9 post-inoculation. Virus titers declined up to the molt, but increased slightly on emergence of adult ticks. Thereafter, virus titers declined progressively, but infectivity could still be detected in adult ticks for up to 205 days post-inoculation. Groups of H. m. rufipes, H. truncatum, and R.e. mimeticus infected adults were fed on susceptible sheep and successfully transmitted CCHF infection. CCHF virus was not isolated from pools of the larval and nymphal progeny of the female ticks nor did the larvae transmit infection to guinea pigs by bite. CCHF virus failed to replicate in adults and nymphae of 3 argasid tick species, Argas walkerae, Ornithodorus porcinus porcinus, and O. savignyi, after intracoelomic inoculation and could be reisolated from the ticks no later than 1 day post-inoculation. The results suggest that all ixodid ticks are capable of transmitting CCHF virus but argasid ticks do not appear to be capable of serving as vectors.

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Viremia and antibody response of small African and laboratory animals to Crimean-Congo hemorrhagic fever virus infection.

Eleven species of small African wild mammals, laboratory rabbits, guinea pigs, and Syrian hamsters were infected with Crimean-Congo hemorrhagic fever (CCHF) virus. Low-titered viremia followed by development of antibody was observed in scrub hares (Lepus saxatilis), Cape ground squirrels (Xerus inauris), red veld rats (Aethomys chrysophilus), white tailed rats (Mystromys albicaudatus), bushveld gerbils (Tatera leucogaster), striped mice (Rhabdomys pumilio), and guinea pigs. The maximum viremic titer in 4 scrub hares was 10(1.7-4.2) 50% mouse lethal doses/ml. Viremia was detected in 1/17 infected laboratory rabbits. Antibody response was only detected in South African hedgehogs (Atelerix frontalis), highveld gerbils (T. brantsii), Namaqua gerbils (Desmodillus auricularis), 2 species of multimammate mouse (Mastomys natalensis and M. coucha), and Syrian hamsters. The results of the study indicate that a proportion of infected scrub hares develop CCHF viremia of an intensity shown in the Soviet Union to be sufficient for infection of feeding immature ixodid ticks, but that South African hedgehogs and wild rodents are unlikely to be of importance as maintenance hosts of the virus in southern Africa.

Animals↗

Inhibition of Crimean-Congo hemorrhagic fever viral infectivity yields in vitro by ribavirin.

Ribavirin was evaluated as a potential therapeutic for Crimean-Congo hemorrhagic fever (CCHF). Viral yields for strains of CCHF virus from Europe, Asia, and Africa in African green monkey kidney (Vero) cells were markedly reduced by this drug. Some CCHF viral strains appeared more sensitive than others, but in general, ribavirin doses as low as 5 micrograms/ml caused a transient reduction of viral yields. A further reduction in viral yields was induced by a dose of 25 micrograms/ml, and evidence of viral replication was not demonstrated in cells treated with 50 or 250 micrograms/ml. In contrast, a dose of ribavirin at least 9 times greater was required to induce a comparable inhibitory effect on the yields of Rift Valley fever virus, for which the drug has been shown to inhibit replication in monkeys and rodents.

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Transmission indices of Loa loa in the Chaillu Mountains, Congo.

A longitudinal entomological survey of the vectors of loiasis was conducted in the Missama area (Lekoumou region) in the Congo from September 1987 to August 1989. The principal catching site was a palm grove surrounded by forest 3 km from the village. Landing/biting densities of Chrysops were measured by standardized fly catches lasting 11 hr carried out twice a month. Vector landing densities were also assessed in the Bantu and Pygmy villages and in the fields. Populations of Chrysops from the palm grove were examined 6 times a month for infection with the infective stage of Loa loa. Chrysops silacea was the predominate vector except at the beginning of the rainy season, when C. dimidiata was the prevailing species. Chrysops were caught throughout rainy season, from October to June. The host-seeking activity of C. silacea was greatest in the middle of this season (February), but occurred sooner (October) for C. dimidiata. The following variables associated with transmission were calculated from our observations in the palm grove (the first figure corresponds to the first year of the study and the figure in parentheses corresponds to the second year). It was calculated that 2.658 (2.185) C. silacea and 1.412 (1.182) C. dimidiata could bite a person in the palm grove per year, including an average of 14.4 (12.7) infective C. silacea and 9.8 (7.2) infective C. dimidiata. The percentage of all dissected flies with third stage larvae in the head and the mean number of larvae in the head/infective fly were 0.57% and 10.1 +/- 6.8 for C. silacea and 0.66% and 11.2 +/- 6.5 for C. dimidiata, respectively. The estimated annual transmission potentials were 171.1 (102.9) for C. silacea and 116.1 (73.8) for C. dimidiata. In the palm grove, transmission was ensured by 2 effective vectors during the rainy season (October to May). Although the annual biting rate for both species was twice as low in the village as in the forest, our data suggest that effective transmission occurs there also.

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Distribution of Crimean-Congo hemorrhagic fever viral antibody in Senegal: environmental and vectorial correlates.

The spatial pattern in Senegal of Crimean-Congo hemorrhagic fever (CCHF) virus IgG antibody prevalence in human and sheep was determined as was the relative abundance of potential tick vectors. A systematic, country-wide serological survey of sheep demonstrated that 10.4% of sheep exhibited IgG to CCHF virus. Sexes were infected equally. Antibody prevalence increased with age from 2.1% during the first year to 18.2% among sheep greater than or equal to 3 years of age. IgG prevalence was highest in the northern, arid Sahelian zone, averaging 75.7% seropositivity, and decreased to zero in the southern, moister Sudano-Guinean and Guinean zones. Human IgG prevalence ranged from 21% to less than 1% among the 8 sites that were sampled throughout the country, being greatest in the arid north and least in the south. Hyalomma ssp. ticks predominated in those biotopes where antibody prevalence was highest. The abundance of Hyalomma ticks may be the proximal determinant of endemic transmission.

Adult↗

Population dynamics of Loa loa and Mansonella perstans infections in individuals living in an endemic area of the Congo.

A follow-up of Loa loa and Mansonella perstans microfilaremia was carried out in an adult population living in a highly endemic area of the Congo. Infection rates and parasite loads were found to be stable in the general population, both in the short-term (two months) and long-term (3-4 years) followup. The microfilarial status of most of the subjects examined did not change between tests. At the individual level, the microfilarial densities of L. loa and M. perstans also remained remarkably constant over time. This results in a qualitative and quantitative stability of the parasitic material available for the vectors.

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Risk of human infections with Crimean-Congo hemorrhagic fever virus in a South African rural community.

Crimean-Congo hemorrhagic fever (CCHF) virus is widely distributed in wild and domestic mammals, birds, and ticks throughout many regions of Africa, Europe, and Asia. Interviews were conducted with 484 individuals from nine farms in the Republic of South Africa from which recent human CCHF cases had originated and with individuals from 27 farms without recognized cases. Serum samples were obtained from all consenting individuals. Blood was also drawn from 2,212 farm animals. Human infection with CCHF virus was uncommon (point prevalence 12.6/1,000). Antibody prevalence in humans on farms increased with age (P less than 0.001), and was correlated with handling lambs. Overall, a greater number of older animals were antibody positive than animals less than one year of age (P less than 0.001), but 12.7% of young animals on farms with human were antibody positive compared with 5.8% on those farms without human infection (P less than 0.05). Physical contact with ticks or tick bite was also found to be a risk factor, but contact with animal blood or fresh meat was not. The risk of CCHF virus infection in the community increased seven-fold following contact with a recognized CCHF case, even when other risk factors were taken into account (point prevalence rate 4.7%). In contrast, antibody prevalence was less than 1% (1 of 128) in the local hospital staff who had cared for patients with CCHF. Prevention is best achieved by education of the farming community and establishing and maintaining awareness in the hospital staff.

Adult↗

Mortality and sequelae due to cerebral malaria in African children in Brazzaville, Congo.

This study was carried out on 170 children admitted to the University Hospital of Brazzaville (Congo) for cerebral malaria between January 1, 1988 and June 30, 1989. The selection criteria were 1) unarousable coma, cerebrospinal fluid without microorganisms or a marked cellular reaction, and the absence of other causes, and 2) that the children lived in Brazzaville. The case fatality rate was 15%. In 75% of the cases, death occurred within the first 48 hr. The prognosis worsened with the stage of the coma and a younger age. At discharge from the hospital, 9% of the cases presented with sequelae. The postcerebral malaria mortality was high; indeed, death occurred in six (7%) of 90 children discharged from the hospital whose parents were contacted between nine and 27 months later. Two deaths were directly related to neurologic sequelae. Among the 58 children examined under satisfactory conditions between nine and 27 months (mean 16.9 months) after discharge, 50% (3 of 6) still presented with attenuated forms of the sequelae observed immediately after the episode of cerebral malaria (cortical blindness had regressed completely, unlike ataxia and loss of balance). Disorders that may have been related to the episode of cerebral malaria were observed in 31% of these 58 cases.

Adolescent↗

Transmission of Crimean-Congo hemorrhagic fever virus in two species of Hyalomma ticks from infected adults to cofeeding immature forms.

Crimean-Congo hemorrhagic fever (CCHF) virus was transmitted from infected adult Hyalomma ticks to uninfected larval and nymphal Hyalomma ticks while cofeeding on a guinea pig host that did not have a detectable viremia. When tested after feeding with infected adults, three (0.8%) of 370 H. truncatum larvae contained detectable CCHF virus (mean virus titer 10(1.6) plaque-forming units [PFU]/tick). The virus was transmitted transstadially from infected larvae and was detected in 15 (1.2%) of 1,253 nymphs and 12 (0.1%) of 2,049 adults. Virus was recovered from 18 (1.9%) of 931 H. impeltatum nymphs, which originated from larvae that cofed with infected adults. After H. impeltatum nymphs cofed with infected adults, CCHF virus was detected in 21 (4.3%) of 449 (mean virus titer 10(1.7) PFU/tick) fed nymphs, but none of 886 adults tested after molt. Results of this study indicate that a small proportion of either larvae or nymphs may acquire CCHF infection while cofeeding on a host without a detectable viremia.

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