The seroprevalences of cystic echinococcosis, and the associated risk factors, in rural-agricultural, bedouin and semi-bedouin communities in Jordan.

An ELISA was used to determine the seroprevalence of cystic echinococcosis (CE), caused by Echinococcus granulosus, in representatives of the rural-agricultural, semi-bedouin and bedouin communities of Jordan. The knowledge, attitudes and practices (KAP) pertaining to the transmission of CE in such communities were also investigated. In the ELISA, serum samples from 2388 subjects were tested for IgG antibodies reacting with antigens in crude sheep hydatid fluid (CSHF). The rural-agricultural subjects were significantly more likely to be seropositive (11.4%) than the semi-bedouin (5.0%) or bedouin (3.7%), but male and female subjects were equally likely to be seropositive. Among the rural-agricultural and semi-bedouin subjects, those aged 11-20 years were most likely to be seropositive. Among the bedouin subjects, however, seroprevalence was highest for those aged 31-40 years. When the distribution of 59 seropositive subjects detected within 36 rural-agricultural households was investigated, 12 (33.3%) of the households were each found to have at least two seropositive members. Immunoblotting indicated that 27.1% of the seropositive rural-agricultural subjects showed immunoreactivity to at least one of the CSHF antigen-B components (of 8-12, 16 and 24 kDa). The living conditions, practices and lifestyles of the rural-agricultural, semi-bedouin and bedouin communities favour the spread of CE in Jordan and warrant an effective programme for the control of the disease.

Cystic echinococcosis in Jordan: socioeconomic evaluation and risk factors.

The costs of illness and surgical intervention for human cystic echinococcosis (CE) cases in Jordan was economically evaluated by 77 surgeons and 77 CE patients. The cost of diagnosis for each CE case was 111.30 US Dollars and 146.20 US Dollars as estimated by surgeons and patients, respectively. The cost of surgical extraction of hydatid cysts for each case was 590.20 US Dollars and 638.50 US Dollars as estimated by both groups, respectively. Knowledge, attitudes and practices (KAP) of 77 CE patients as well as several Jordanian groups with different occupations including 144 shepherds, 119 settled livestock owners, 25 slaughter house workers, 400 university students and 80 inhabitants of a CE focus in southern Jordan were analyzed through a set of questionnaires. All of these groups had poor knowledge of CE, especially the source and causes of infection. All practices and attitudes of each group favored continuous transmission of the parasite and indicate the need for the implementation of a proper control program in the country.

Human cystic echinococcosis in the West Bank of Palestine: surgical incidence and seroepidemiological study.

The surgical incidence of cystic echinococcosis (CE) due to Echinococcus granulosus was investigated in hospitals of the West Bank, Palestinian Authority between January 1990 and December 1997. Serum samples from school-children in Yata town, which showed the highest surgical incidence, were tested for anti-hydatid antibodies. A total of 390 surgically confirmed cases were recorded throughout the 8-year period, with an overall mean annual surgical incidence (MASI) of 3.1 per 100,000. A high MASI of 4.9, 5.0 and 5.1 per 100,000 was found in Hebron, Jericho and Bethlehem Governorates, respectively. Yata town, Hebron governorate, showed the highest MASI, at 16.8 per 100,000. The highest incidence was found in age groups 11-20 and 21-30 years, at 27.4% and 21.5% of the total number of cases. While there was no significant gender difference in the number of cases in the age groups of 20 years or less, the male to female case ratio was 1:3.2-4.1 in the older age groups. The liver was the most common site of hydatid cysts in 69.9% of cases. Lung cysts were predominant in younger age groups (20 years or less). The seropositivity for CE in the school-children of Yata was 2.4% and 2.1% using enzyme-linked immunosorbent assay and the indirect haemagglutination test, respectively. CE is a significant endemic disease throughout the West Bank. The disease is acquired early in life and is more prevalent among females than males. Behaviour and life-style favour the spread of the disease.

Diagnosis of canine echinococcosis: comparison of coproantigen detection with necropsy in stray dogs and red foxes from northern Jordan.

The sandwich enzyme linked immunosorbent assay (ELISA) was used as a diagnostic test for Echinococcus granulosus infection by detecting coproantigens in 94 stray dogs Canis familiaris and eight red foxes (Vulpes vulpes) from northern Jordan. The results were analyzed in relation to actual helminth infection as revealed by necropsy. The infection rate of dogs with E. granulosus was 13.8% with a worm load ranging between 3-> 10,000 per infected dog. In contrast, eight of 13 E. granulosus infected dogs were coproantigen positive (overall sensitivity 61.5%). The sensitivity increased to 87.5% and 100% in dogs harboring > 20 and > 100 worms/dog, respectively. The specificity of coproantigen-ELISA was 91%. The greatest cross-reactivity was found in dogs infected with Dipylidium caninum. The positive and negative predictive values for the coproantigen-ELISA test were 50% and 94.2%, respectively. Thus, a coproantigen negative dog is most probably truly negative for E. granulosus. In contrast, a coproantigen positive dog may not be truly positive for E. granulosus, except if it has a high worm burden of > 100 worms/animal.

Prevalence of intestinal helminths of dogs and foxes from Jordan.

Necropsy of 340 stray and semi-stray dogs (Canis familiaris) and nine red foxes (Vulpes vulpes) from Jordan revealed that 239 dogs (70.3%) and all foxes were infected with at least one intestinal helminth species. No trematodes were found in the intestine of these hosts. The overall infection rates with cestodes, nematodes and acanthocephalans in dogs were 66.8%, 4.4% and 2.9%, respectively. The following cestodes were identified: Echinococcus granulosus (9.4%), Taenia pisiformis (11.8%), T. hydatigena (7.4%), T. ovis (4.4%), T. multiceps (3.8%), T. taeniaeformis (2.9%), Dipylidium caninum (19.4%), Joyeuxiella (3.2%), Diplopylidium (2.4%), and Mesocestoides (0.9%). Other intestinal worms in dogs were Toxascaris (2.6%), Toxocara canis (1.2%), and Protospirura (0.6%) nematodes, and gigantorhynchiid acanthocephalans (2.9%). Intestinal helminths found in foxes included cestodes (D. caninum, Joyeuxiella, Diplopylidium, Mesocestoides), nematodes (Protospirura, Uncinaria stenocephala and Oxynema) and an acanthocephalan (Macracanthorhynchus). In both hosts, most helminths were recovered from the second intestinal segment of four equally divided segments.

Differential serodiagnosis for cystic and alveolar echinococcosis using fractions of Echinococcus granulosus cyst fluid (antigen B) and E. multilocularis protoscolex (EM18).

Echinococcus granulosus cyst fluid and E. multilocularis protoscolex extract were fractionated by a single step of preparative isoelectric focusing, resulting in an antigen B-rich fraction (8-kD) and an Em18-rich fraction, respectively. The usefulness of both fractions for differential serodiagnosis of cystic (CE) and alveolar (AE) echinococcosis was evaluated by a large-scale immunoblot analysis on a battery of 354 serum samples. These included 66 from AE patients originating from four different endemic areas, 173 from CE patients originating from seven different endemic areas, 71 from patients with other parasitic diseases, 15 from patients with hepatomas, and 29 from healthy individuals. In an immunoblot with the antigen B-rich fraction, 92% (158 of 173) of the CE sera as well as 79% (52 of 66) of the AE sera reacted with the 8-kD subunit. No cross-reactivity occurred with any sera from patients with cysticercosis, other parasitic diseases, or with hepatomas, or from healthy controls. In an immunoblot with the Em18-rich fraction, all but two sera from AE patients (64 of 66, 97%) recognized Em18, and only nine of 34 CE sera from China reacted with it. All other (139) CE sera from six other countries were negative as were all (115) other non-echinococcosis sera. These findings indicate that antigen B (8-kD) is not species-specific for E. granulosus but is genus-specific for Echinococcus, and that the Em18 antigen is a reliable serologic marker for species-specific differentiation of AE from CE.

Short report: outbreak of cutaneous leishmaniasis in a nonimmune population of soldiers in Wadi Araba, Jordan.

An outbreak of zoonotic cutaneous leishmaniasis (ZCL) occurred in a battalion of 80 soldiers posted at Qurayqira camp in Wadi Araba in southern Jordan. The battalion spent an intermittent period of five and a half months in the area, during which 45.0% (36 of 80) of the soldiers showed clinical disease. Of the 44 clinically negative soldiers, 31 were tested with leishmanin and 11 (35.5%) were leishmanin positive. The number of lesions in infected soldiers ranged from one to 15 and were mostly on the face and extremities. This report shows the level of transmission of ZCL in Wadi Araba, which is presently undergoing economic expansion and development following the peace process of the Arab-Israeli conflict.

A new focus of cutaneous leishmaniasis caused by Leishmania tropica in northern Jordan.

An endemic focus of cutaneous leishmaniasis caused by Leishmania tropica was identified in the district of Bani Kinana, northern Jordan. Fifty-five autochthonous cases were detected from February to September 1994 in a population of about 50,000. Most cases (74.5%) were from the villages of Malka and Um-Quais. The lesions were typically small, dry and mostly located on the face and extremities. Multiple lesions were common. Incidence was higher in females than in males and familial clustering of cases was observed in houses on the outskirts of villages. A leishmanin survey showed a 23.3% positivity rate in the populations of Malka and Um-Quais. The rate of positivity was low in the younger age groups and increased with age. There was some evidence that the disease is a zoonosis in this focus.

In vitro cultivation of Trypanosoma acomys: production of insect stages and bloodstream forms.

When Trypanosoma acomys bloodstream forms were cultivated at 37 degrees C in Schneider's Drosophila medium supplemented with 20% (v/v) heat-inactivated foetal calf serum (FCS), with Microtus agrestis embryonic fibroblasts in RPMI 1640 medium supplemented with 20% FCS or in Baltz's medium supplemented with 10% FCS, the parasites transformed and largely remained as epimastigotes. Epimastigotes were also usually the commonest stage observed when the parasites were co-cultivated with a mosquito cell line at 27 degrees C. However, if these cultures were initiated with the supernatant suspensions from fibroblast cultures that had been cryopreserved, trypomastigotes, including bloodstream-like forms, were the predominant stage for the first 4 days of culture. It is suggested that the glycerol supplement or the temperature changes stimulated this unusual morphogenesis. At 27 degrees C, T. acomys was incapable of multiplying and died when cultured in fresh Schneider's Drosophila medium supplemented with 20% FCS, but co-cultivation with the mosquito cell lines or cultivation in cell-free supernatants from 1-week-old mosquito cell cultures was successful at this temperature; most of the parasites multiplied as epimastigotes.

Serodiagnosis and seroepidemiology of human unilocular hydatidosis in Jordan.

A total of 2182 serum samples from 38 patients with surgically confirmed unilocular hydatidosis, 19 clinically assessed patients, 15 patients with parasitic infections other than hydatidosis, 104 hospital outpatients, and 2006 normal Jordanians were serodiagnosed for the presence of IgG antibodies against hydatid fluid, circulating immune complexes (CIC), and/or hydatid circulating antigen (CA). Anti-hydatid IgG antibodies were detected in the sera of 77.4% of patients with hydatid disease and persist for very long periods postsurgery. As many as 54.1% of patients with hydatidosis had positive levels of CIC, and 16.1% had circulating antigen in their sera. The search for circulating antigen and CIC decreased the number of false-negative hydatid cases from seven to three, and the combined sensitivity of the assays thus increased from 77.4% to 90.3%. Using the immunoblot technique, 16- and < 14.4-kDa Echinococcus granulosus-specific bands were detected in sera from 54.1% and 61.5% of patients with hydatid disease who were tested before and after surgery, respectively. The seropositivity rate for anti-hydatid IgG antibodies was 2.4% for the general Jordanian population and 5.8% for hospital outpatients.

Echinococcus granulosus: possible formation of a shelled egg in vitro.

What appeared to be the early stages in the formation of a single egg with a striated embryophore was observed in an in vitro culture of Echinococcus granulosus protoscoleces isolated from sheep hydatid cysts in North Jordan. The 'egg' measured 19 x 19 microns in diameter and was formed in an intermediate vesicular/monozoic form which was never previously reported from a culture. This is the first report of an apparently shelled egg forming in an in vitro culture, but although promising, cannot be regarded as being unequivocal and will require confirmation by further work.

In vitro culture of the strobilar stage of Echinococcus granulosus of sheep and donkey origin from Jordan.

Protoscolices of Echinococcus granulosus isolated from hydatid cysts of sheep and donkeys in Jordan were cultured in vitro using a modified diphasic culture system. Protoscolices from these two sources manifested differences in the mode of development, evagination and growth rates. Protoscolices isolated from sheep cysts grew in vitro in the polyzoic direction up to the three- to four-segmented mature worms, reaching a length of 2.9 mm. In contrast, donkey protoscolices failed to develop beyond the early stages, even after 67 days of culturing. On prolonged culturing, few worms of donkey origin reached the banding and segmentation stages, attaining a maximal length of 1.6 mm at periods ranging between 81 and 114 days of culturing. None of these segmented worms showed genital differentiation or analgen. The evagination and growth rates of protoscolices isolated from donkey liver cysts were compared with those obtained from sheep liver or lung cysts. The most significant difference in these rates occurred at the commencement of the segmentation stage. Differences in the development, growth and evagination rates observed between the donkey and sheep forms may reflect the strain variation of E. granulosus in this country.

Urbanization--how does it affect the behaviour of sandflies?

The effect of building activities on the abundance of sandflies was investigated in Jordan. A total of 3442 sandflies were collected. The largest collections were made from natural habitats, uninhabited and inhabited houses. Collections from the latter two often included a larger number of Phlebotomus species. It was evident from this study that sandfly populations are able to recover to, and often exceed their original natural densities in man-made artificial habitats.

The behaviour and dispersal of sandflies in Ras el Naqb, south Jordan with particular emphasis on Phlebotomus kazeruni.

The behaviour and dispersal of sandflies from an enclosed cave were investigated during the summer of 1988 in Ras el Naqb by the mark-release-recapture technique. Nine species of sandflies were reported during this study including 5 Phlebotomus and 4 Sergentomyia species. P. sergenti, P. alexandri, S. adleri/S. clydei and S. taizi were reported for the first time from Jordan. P. kazeruni was the most abundant sandfly species inside the cave. The maximum distance travelled by a single P. kazeruni was 340m. However, the majority were recaptured within 10m of the cave. It seems likely that an independent "population" of sandflies which breeds inside the cave is developing.

Trypanosoma acomys (Wenyon, 1909): continuous culturing with a mouse fibroblast cell-line (A9).

The continuous culturing of Trypanosoma acomys in the presence of a murine areolar-adipose cell line (A9) was possible for the 1st time. The trypanosomes were cultured at 37 degrees C with A9 in DMEM supplemented with 20% heat inactivated fetal bovine serum, using an initial inoculum from primary cultures of lung or blood clots from infected spiny mice. The cultures were maintained for 115 days and underwent 15 passages before termination and cryopreservation. Using this culture system T. acomys subcultures were initiated from 3 different initial inocula (3 x 10(4), 1.5 x 10(5) and 7.4 x 10(5) parasites/ml) and growth curves revealed that the lowest inoculum gave the best growth pattern. This inoculum yielded a population doubling time of less than 12 h for 4 days, a high peak density of 7 x 10(6) parasites/ml and the most gradual decline compared to the other 2 inocula. Rosetting epimastigotes and nests of amastigotes were observed in close association with the feeder layer cells. Epimastigotes were the most predominant form in culture supernatants but other morphological forms observed included trypomastigotes and sphaeromastigotes.

Spleen hydatidosis in sheep from north Jordan.

A total of 380 ewes of different age groups were inspected for spleen hydatidosis during the summer of 1985. Of these, 7.6% had cysts in the spleen. There was an increase in the prevalence of the disease with increasing age of the animal, reaching 11.8% in greater than or equal to 8-year-old ewes. Out of all the animals with spleen hydatidosis, 61.5% were found to have fertile cysts and, when classified, 46.7% were found to be fertile with a mean total number of protoscolices of 13.4 x 10(3) per cyst. The majority of the protoscolices were found to be viable (74%). Spleen hydatidosis which is mainly solitary seems to be secondary to the involvement of the liver or the liver and lung simultaneously. The importance of these aspects of spleen hydatidosis are discussed.

Trypanosoma acomys (Wenyon, 1909): reproductive forms and course of parasitemia in the natural host Acomys cahirinus (Desmarest, 1819).

Trypanosoma acomys (Stercoraria, Trypanosomatidae) was studied in the laboratory-bred spiny mouse Acomys cahirinus. Only trypomastigotes were found in peripheral blood, whereas the thymus of experimentally infected A. cahirinus showed amastigote syncytial reproductive forms whose nuclei of nuclei ranged from 2 to 128. These reproductive forms were detected 9-10 days after the i.p. injection of spiny mice with trypanosomes obtained from primary lung cultures. Transitional trypomastigote-like forms were observed in the thymus; no reproductive or diving forms were encountered in any other tissue, including blood. The course of the parasitemia in spiny mice inoculated i.p. with 1.2 x 10(6) parasites from 26- to 32-day-old cultures initiated from infected lungs was studied. A similar, characteristic pattern was observed in both male and female hosts. Four phases were recognized: latent, logarithmic, stationary, and decline. These phases ended on days 2, 14, 98, and 154 postinoculation, respectively. The decline phase was gradual and the parasites could not be detected by day 154 postinoculation. In contrast, inoculation from older cultures (35-50 days old) showed longer latent and logarithmic phases, a shorter stationary phase, and a sharp drop in parasite density during the decline phase; the parasites were not detected in the peripheral blood by 120 days postinoculation.

The immune response to Schistosoma mansoni infections in inbred rats. VI. Regulation by T cell subpopulations.

These studies assess the roles of subpopulations of T lymphocytes in inducing and modulating resistance to Schistosoma mansoni. CDF rats were depleted of RT 7.1+ (anti-Pan-T), W3/25+ (anti-T helper/inducer), or OX8+ (anti-T suppressor) cells by the in vivo administration of monoclonal antibodies (mAb). The development of parasites and immunity to challenge by S. mansoni were compared with results in undepleted normal and congenitally athymic rats. Discrete subpopulations of T lymphocytes were adoptively transferred to ascertain effects upon parasite development and the protective immune response. In vitro studies, involving utilizing cocultivation of cell subpopulations +/- cyclosporin A, were utilized to dissect mechanisms. Depletion of T lymphocytes by anti-RT7.1 mAb and anti-W3/25 mAb resulted in augmented initial worm development, suboptimal resistance, and decreased antibody and delayed-type hypersensitive reactivity directed against schistosome antigens. Depletion with OX8 mAb produced opposite effects. The adoptive transfer of T cell subpopulations produced concordant results with T cell regulation expressed B cell-dependent effector mechanisms. The coadoptive transfer of cells resulted in the suppression of resistance afforded by the W3/25+ cells by OX8+ cells, which could be augmented in vitro by cyclosporin A. Thus, protective immunity to S. mansoni in rats is regulated by discrete subpopulations of T lymphocytes. The findings suggest the possibility of selective immune regulation of resistance based on the manipulation of specific T cell subpopulation.