IgG4 serology of loiasis in three villages in an endemic area of south-eastern Gabon

Human filariasis due to Loa loa differs from other filariasis in that the majority of infected subjects are without circulating microfilariae (occult loiasis). In search for alternative diagnostic methods, which do not depend on circulating microfilariae or the (rather infrequent) eye-passage of adult worms, it was shown earlier that IgG4 antibodies directed against Loa loa adult worm antigen are apparently a good marker of occult loiasis and specific with regard to the sympatrically occurring Mansonella perstans . In this study we evaluated an IgG4 antibody-based ELISA using crude extract of Loa loa microfilariae (which is easier to obtain than adult worm) to estimate the prevalence of loiasis in 3 villages in South-East Gabon. Of 222 examined individuals (80 children < 16 years, 142 adults) 44 (20%) carried Loa loa microfilariae and 170 (77%) M. perstans . Using the mean OD-value + 1 standard deviation of 9 sera from patients solely infected with M. perstans (from the Gambia, where Loa loa is not endemic) as a cut-off, 35 of the 44 microfilaraemic Loa loa patients and 2 of the 9 Gambian controls were positive. This shows that our method had a sensitivity of 80% and a specificity of 78%. Among the remaining 178 subjects who had no microfilariae of Loa loa , as many as 97 (55%) had significant levels of specific IgG4 antibodies against Loa loa , suggesting that they carried occult loiasis. The mean IgG4 level in these putatively occult loiasis patients was slightly but significantly lower than in microfilaraemic subjects ( P < 0.03). In conclusion, despite the limited sensitivity and specificity of our method, IgG4- ELISA at present is a very useful tool in estimating the real prevalence of loiasis in epidemiological surveys and at the individual level can confirm the diagnosis of L. loa amicrofilaraemic subjects with clinical signs suggesting loiasis.     

Serum immunoglobulin G4 antibodies to the recombinant antigen,Ll-SXP-1, are highly specific for Loa loa infection

The clinical manifestations and geographic distribution of loiasis overlap with those of other human filarial parasites, presenting challenges in the specific diagnosis of loiasis that may lead to delays in appropriate therapy. A recombinant antigen (Ll-SXP-1), preferentially recognized by serum samples from experimentally infected rhesus monkeys, was identified from a Loa loa L3 cDNA library. IgG4 antibody reactivity to purified Ll-SXP-1 was assessed by means of ELISA, using serum samples from patients with loiasis, lymphatic filariasis, onchocerciasis, mansonellosis, or other helminthiases and healthy control subjects. The assay was 56% sensitive and 98% specific for loiasis. Antibody reactivity was detectable before microfilaremia in experimentally infected rhesus monkeys and declined (but did not disappear) after diethylcarbamazine therapy in infected patients. IgG4 antibodies to recombinant Ll-SXP-1 are a highly specific marker of L. loa infection and may be useful for the diagnostic evaluation of persons with filariasis of unclear etiology.     

Expression of filarial-specific IgG subclasses under different transmission intensities in a region endemic for loiasis

Specific IgG subclasses were investigated in two villages (Okoumbi and Ndjokaye) in southeast Gabon with different Loa loa transmission intensities of approximately 9,000 and 1,300 infective larvae (L3) per person per year, respectively. IgG subclasses were measured by an enzyme-linked immunosorbent assay (ELISA) using extracts of L. loa L3, microfilariae (MF), or adult worms. Levels of L3-specific IgG3 were significantly higher in the village with low transmission (Ndjokaye) (P = 0.006). In contrast, MF-specific IgG2 was significantly higher in Okoumbi than in Ndjokaye (P = 0.0009). In the high-transmission village (Okoumbi), levels of both MF- and adult-specific IgG4 were significantly increased in MF carriers compared with amicrofilaremic subjects (P = 0.0015 and P = 0.003, respectively), while levels of L3- and MF-specific IgG1 were significantly higher in amicrofilaremic individuals compared with MF carriers (P = 0.04 and P = 0.03, respectively). Furthermore, among microfilaremic individuals, the level of the specific IgG1 subclass was much lower in Okoumbi than in Ndjokaye (P = 0.036). These results suggest that the expression of antigen-specific IgG3 and IgG2 is more likely to vary with transmission intensity, whereas antigen-specific IgG4 and IgG1 varies with adult worm and MF burden.     

Prevalence of filariasis in symptomatic patients in Moyen Chari district, south of Chad

Filarial parasites infect an estimated 140 million people worldwide. Wuchereria bancrofti, Onchocerca volvulus, Loa loa and Mansonella perstans are responsible for most filarial infections in sub-Saharan Africa. We describe the prevalence and the clinical characteristics of filariasis in symptomatic patients in Goundi Sanitary district:167 patients were enrolled (99 men, 68 women). M. perstans microfilariae were isolated in peripheral blood in 164 cases, while Loa loa and Wuchereria bancrofti filariasis were diagnosed in only six and three cases, respectively. The most frequent filariasis observed in our study were due to M. perstans and L. loa, while the few cases of W. bancrofti filariasis seem to have been acquired abroad. No cases of O. volvulus were observed. Microfilarial burden was not related to symptoms, but a correlation between eosinophilia and pruritus was evident. No relationship was observed between eosinophils and symptoms. The prevalence observed in symptomatic patients could reflect the real prevalence of filariasis.     

Filariasis in the Igwun River Basin, Nigeria: an epidemiological and clinical study with a note on the vectors

In a cross-sectional, epidemiological and parasitological study of human filariasis, 845 individuals were examined in settlements along the Igwun Basin, Imo State, Nigeria. Four different filarial nematode species were identified. Two hundred and fifty-six (30.3%) of the individuals examined were positive for Onchocerca volvulus, 113 (13.4%) for Mansonella perstans, 76 (9.6%) for Wuchereria bancrofti and 77 (9.1%) for Loa loa. Microfilarial rates increased with age of individuals and showed a tendency towards higher prevalence rates in males than in females. The intensity of O. volvulus infection was high, with the highest microfilarial density of 44 mf mg-1 snip which occurred in the 40-49-year-old individuals. In W. bancrofti and L. loa infections, infections of over 1000 mf 20 ml-1 blood were recorded in 15.8% and 19.5% of individuals, respectively. Observed clinical signs were associated with inflammatory, lympho-obstructive and ocular manifestations. In M. perstans infections all clinical cases were inflammatory. In W. bancrofti, 44.4% of clinical cases were inflammatory, and lympho-obstructive manifestations consisted of 23.8% chyluria, 12.7% hydrocele and 19.1% elephantiasis. In L. loa infections all clinical cases were inflammatory with indications of Calabar swellings. In O. volvulus infections 23.5% of clinical cases were inflammatory, while 76.5% showed ocular manifestations. The absence of blindness despite high O. volvulus infection rates was remarkable. The presence of potential insect vectors and the occurrence of clinical signs are indications of active transmissions.     

Case report: Loiasis with peripheral nerve involvement and spleen lesions

Loiasis, which is caused by the filarial nematode Loa loa, affects millions of persons living in the rainforest areas and savannah regions of central Africa. Typical manifestations are calabar swellings and the eyeworm. We report a case of loiasis with unusual clinical complications: a peripheral neuropathy and focal hypo-echogenic lesions of the spleen, which disappeared after treatment with albendazole and ivermectin. The literature reports that L. loa infection can be associated with various manifestations, some of them being serious. More information is needed to better characterize the protean manifestations of the disease in loiasis-endemic areas to evaluate the true incidence of loiasis.     

High prevalence of occult hepatitis B in Baltimore injection drug users

Occult hepatitis B is defined by the presence of hepatitis B virus (HBV) DNA in a serum or liver in the absence of hepatitis B surface antigen (HBsAg). The prevalence and clinical correlates of occult hepatitis B remain incompletely defined. A cross-sectional study was performed to determine the prevalence of occult hepatitis B in a high-risk cohort composed of 188 injection drug users in Baltimore, Maryland. All individuals had chronic hepatitis C viral infections confirmed by RNA detection and liver biopsy. Serologic assays for HBsAg and core antibody (HBcAb) were performed. Serum HBV DNA was detected using the COBAS HBV AMPLICOR monitor assay (lower limit of detection, 200 HBV copies per milliliter) and a semi-nested polymerase chain reaction (PCR) assay (lower limit of detection, 15 HBV copies per milliliter). Although almost all individuals (96%) were anti-HBC positive, only 8 of 188 (4%) were HBsAg positive. Occult hepatitis B was not identified using the COBAS assay, but was found in 81 of 180 (45%) of individuals using semi-nested PCR. Of the 8 HBsAg positive individuals, HBV DNA was found in 1/8 using the COBAS assay and 6/8 using the nested PCR assay. Overall, liver disease was mild, with a median serum alanine aminotransferase (ALT) of 38 IU/L, median activity grade of 3/18, and median fibrosis stage of 1/6. No association was found between the serum AST (aspartate aminotransferase), activity grade, or stage of liver disease and the presence of occult hepatitis B. Serum ALT levels were slightly higher in patients without occult hepatitis B (46 vs. 35 IU/L), and the median years since first injection drug use was somewhat longer in those without occult hepatitis B (24 vs. 20 years). In conclusion, although further research is needed to assess its clinical significance, there is a high prevalence of occult HBV infection in this cohort of HCV-infected injection drug users.     

The use of mebendazole in the treatment of filariases due to Loa loa and Mansonella perstans

Patients harbouring Loa loa and/or Mansonella perstans infections, or presenting typical filarial symptomatology, were treated with mebendazole, a drug shown to be efficient in other parasitoses. Of the patients who took the drug during 21 days and provided regular blood samples, 84% showed a significant decrease in their L. loa microfilaraemia and 100% in their M. perstans microfilaraemia, with a concomitant improvement in the clinical manifestations and no side reaction to drug administration. Additionally, the long-lasting effect of the drug (up to day 200 in some cases) may suggest a positive action on the adult worm. Mebendazole may consequently be regarded as a safer alternative to diethylcarbamazine for the treatment of these two parasitoses.     

Genetic epidemiology of host predisposition microfilaraemia in human loiasis

Evidence is accumulating from experimental and human studies that genetic factors are involved both in the control of infectious diseases and in the regulation of infection levels and clinical presentation. So far few studies have investigated the role of these genetic factors in human infection by the filarial parasite Loa loa. We present a segregation analysis on 74 nuclear families who live in the tropical rainforest of southern Cameroun and are exposed to homogeneous loiasis transmission. The results indicate that there is a genetic predisposition to be microfilaraemic and that predisposed subjects might be genetically unable to mount an efficient immune response against loiasis antigens. This individual susceptibility could explain at least in part why the prevalence of infection (microfilaraemic individuals) does not usually exceed 30% of the exposed population in hyperendemic regions. Further genetic studies, based on linkage analysis using both familial information and genetic markers, will help to identify the nature of the genetic factors predisposing to microfilaraemia.     

慢性乙型肝炎病毒感染者家族隐匿性乙型肝炎病毒感染的调查

目的 了解慢性HBV感染者家族隐匿性HBV感染的发生率及其与HBV标志物、年龄和性别等的关系.方法 ELISA方法检测慢性HBV感染者家族成员的HBV血清学标志物,套式PCR法检测136例HBsAg阴性家族成员的血清HBV DNA,并将隐匿性HBV感染者和HBsAg、HBV DNA均阴性者分别作为试验组和对照组进行HBV标志物、年龄、性别和生物化学检测结果的比较.两组均数比较采用t检验.率的比较采用χ~2检验或Fisher确切概率法检验.结果 在52个慢性HBV感染者家族中共检测到92例HBsAg阳性者和136例HBsAg阴性者,其中15例为隐匿性HBV感染者,慢性HBV感染者家族HBsAg阳性率和隐匿性HBV感染的发生率分别为40.4%和11.0%,15例隐匿性HBV感染者中有7例抗-HBc阳性(χ~2=5.341,P=0.02),但隐匿性HBV感染的存在与年龄、性别等无关.结论 HBV感染存在家庭聚集现象,且在其家族中存在隐匿性HBV感染,并在抗-HBc阳性者中发生率较高.     

Clinical characteristics of post-treatment reactions to ivermectin/albendazole for Wuchereria bancrofti in a region co-endemic for Mansonella perstans

Post-treatment reactions to single-dose ivermectin (200 microg/kg) and albendazole (400 mg) were studied in a filarial endemic region of Mali. The prevalence of Wuchereria bancrofti in this region was 48.3% (69 of 143), and coinfection with Mansonella perstans was common (30 of 40, 75%). Microfilarial levels of M. perstans correlated positively with age (P = 0.006) and with W. bancrofti microfilarial levels (P = 0.006). Forty individuals (28 infected and 12 uninfected) were treated, with mild post-treatment reactions occurring in 35.7% (7 of 28) of the W. bancrofti-infected subjects. Reaction severity correlated with pretreatment W. bancrofti microfilarial levels (P = 0.001). There were no significant differences in the prevalence or severity of post-treatment reactions in those who were co-infected with M. perstans. It is concluded that co-infection with M. perstans does not significantly alter the post-treatment reaction profile to single-dose ivermectin/albendazole in W. bancrofti infection in this community, and that acute post-treatment reactions should not limit patient compliance in community-based programs to eliminate lymphatic filariasis.     

Experimental infection of humans with filariae

This report summarizes the findings of the 17 published studies involving humans who have been experimentally infected with filarial parasites. Over the past 60 years, 45 individuals have been deliberately infected with Wuchereria bancrofti, Brugia malayi, Brugia pahangi, Loa loa, Mansonella perstans, Mansonella ozzardi, and/or Onchocerca volvulus. The findings from these experimental infections of humans have helped define microfilarial survival and periodicity within human hosts, the prepatent period for the causative agents of lymphatic filariasis, etiologic agents for particular clinical syndromes, immunologic and hematologic consequences of filarial infection, and the role of chemotherapeutic agents in the prevention and treatment of filarial infections.     

Chrysops silacea biting densities and transmission potential in an endemic area of human loiasis in south-west Cameroon

We studied the biting densities of Chrysops silacea and the transmission of loiasis over 1 year in a regenerated forest in the south-west province of Cameroon. A total of 3015 flies caught near a wood fire at ground level during rainy and dry seasons were identified morphologically and 1975 caught during the rainy season were dissected to determine their physiological age and infection rate. The prevalence of microfilaraemia in the human population in the study area was determined using the thick blood smear method. Chrysops silacea was the only species caught. The daily and seasonal biting cycle of C. silacea showed two peaks of activities, 9-11 a.m. and 2-4 p.m. The biting cycles of parous and nulliparous flies showed the same trends, but the density of nulliparous flies biting at all time of the day was 2-3 times higher. Chrysops silacea biting density was high during the rainy season (9.06 +/- 6.88 flies/man/h) and lowest during the dry season (0.44 +/- 0.75 flies/man/h). An infection rate of 1.72% and a monthly morning and afternoon transmission potentials of 120769.11 and 139016.64 infective head L3/man were observed, respectively, in the rainy season. Even though few Chrysops carried Loa loa infective larvae (0.7%), their parasite load was high, giving a high level of transmission of L. loa in the area. A total of 20.37% of the people examined for blood microfilariae were positive. These results suggest that the study area is an active focus of loiasis transmission.     

The migration and localization of Loa loa infective and fourth-stage larvae in normal and immunosuppressed rodents

The migration and localization of the human filarial parasite Loa loa in laboratory mice (BALB/c and Swiss) and jirds (Meriones unguiculatus) was investigated. The rodents, either left immunocompetent or immunosuppressed with hydrocortisone, were each inoculated subcutaneously or intraperitoneally with 50 or 200 infective, third-stage larvae (L(3)) of L. loa. Groups of the rodents were killed at various times post-infection, up to day 40, to enable histological studies and permit developing larvae to be recovered. Larvae survived and developed for only 1 week in the immunocompetent rodents but for a mean of 3 weeks in the immunosuppressed. Most of the larvae were found in the subcutaneous tissues (81.9%), peritoneal cavity (14.9%), pleural cavity (1.8%) or the lungs and heart (1.3%) and none was detected in the spleen, kidney, intestine, liver or pancreas. Localization of the larvae appeared unaffected by the site of inoculation, the rodent species or strain, or the dose of L(3) used. The recovery of larvae (as a percentage of the number inoculated) was better among the rodents inoculated with 50 L(3) each than among those given four times as many L(3). The results of the histological studies not only confirmed the presence of larvae in the subcutaneous tissue (72.5%), muscles (11.7%) and peritoneal and pleural cavities (7.8%) of the infected rodents but also revealed worms in the lymphatic vessels of the mesentery and spinal cord (7.3%). These results indicate that most L. loa L(3) inoculated into a mammalian host localize in the cutaneous sites and that only a small proportion of them might migrate, using the lymphatic system, into the internal organs. The observation of migrating L. loa larvae in the lymphatic vessel of the meningeal envelope of the spinal cord, albeit in an experimental host, may explain why, in areas where human loiasis is endemic, neurological manifestations occasionally occur in those with L. loa infections.     

Overt and occult hepatitis B virus infection among community children in Northwest China

Although a universal newborn hepatitis B (HB) immunization programme has been implemented in China, hepatitis B virus (HBV) breakthrough infection, including HB surface antigen (HBsAg)‐positive infection and occult HBV infection (OBI), still occurs during infancy or childhood. Obtaining the actual prevalence of HBV infection in general children is important for preventing and controlling the spread of HB. Accordingly, we investigated the prevalence of overt infection and OBI in community children and compared the serological and virological characteristics of OBI and HBsAg carrier children to clarify the mechanisms related to OBI. In total, 6 706 community children <12 years of age were included from a population‐based HBV seroepidemiological investigation in Northwest China. The HBsAg carrier rate in community children was 1.60% (107/6706), and the anti‐HBs positive rate was 57.35% (3846/6706). Additionally, 1192 HBsAg‐negative children were examined for OBI using nested PCR. The prevalence of OBI in local children was 1.26% (15/1192), and the predominant OBI genotypes were C and D. The 15 OBI children and 29 HBsAg‐positive children from the same population did not have a statistical significant difference in age, gender, alanine aminotransferase (ALT), proportion of anti‐HBs or anti‐HBc, viral genotypes or mutations. Children with chronic overt infection had higher viral loads than OBI children (P=.004). These results suggested that HBV overt and occult infection of children was more serious in underdeveloped north‐west regions. HBV neonatal immunization and catch‐up programmes should be strengthened and supplemented. None of specific viral mutations or genotypes related to OBI were found. OBI may be a specific stage of HBV infection.     

Comparison of thick blood smear and saponin haemolysis for the detection of Loa loa and Mansonella perstans infections

In an endemic area, Loa loa and Mansonella perstans microfilariae were detected by the examination of 5 ml blood in respectively 7.4 and 26.2% of subjects who would have been erroneously considered amicrofilaraemic by the conventional method of two thick blood smears (40 microliters blood). Correction factors to be applied to the results obtained with thick blood films in order to approach the true parasitological prevalences were 1.5 for L. loa and 1.6 for M. perstans. In addition, the analysis of a large volume of blood provides a better estimation of microfilarial load of the parasitized population.     

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.     

Diagnosing multiple parasitic infections: trypanosomiasis, loiasis and schistosomiasis in a single case.

A case is reported of a 32-year-old traveller with loiasis, schistosomiasis and African trypanosomiasis. The patient had been working in oil exploration in Nigeria and Gabon and presented with Calabar swellings and carpal tunnel syndrome. Serology for all 3 diseases was positive but microfilariae of Loa loa and ova of schistosomiasis were not found. Treatment with diethylcarbamazine and praziquantel was given for loiasis and schistosomiasis respectively. Trypanosomes were isolated from a lymph node aspirate only after repetition of the procedure 2 months later and the patient was treated with suramin. He developed a drug induced nephritis and was then treated successfully with alpha-difluoromethylornithine. There is a discussion of the difficulties encountered making these diagnoses in Europeans particularly where there are atypical clinical features. The risks of rural work in West Africa are noted and the importance of considering all parasitic diseases relevant to the travel/occupational history is emphasised.     

A case of loiasis

Loiasis is quite common in the endemic regions of Central and West Africa. But only three cases were reported in Japan. This is a report of a 28 year old male from Gabon infected with Loa loa with eye symptoms as the chief complaint. For the first time in Japan he was treated with Ivermectin (IVM) which is recently attracting attention as the drug for filariasis world wide. IVM therapy was effective, and decreased the counts of microfilarias in the patient's blood. No adverse effect was seen in this patient. This case suggested that IVM is an useful drug for loiasis, and further study is warranted.