Microfilaruria in onchocerciasis. A clinical and epidemiological follow-up study in the Republic of Chad

Microfilariae of O. volvulus were recovered from the urine of 11% of the residents of a village in the Republic of Chad where onchocerciasis was holoendemic. A follow-up study of the same population 3 years after the original investigation revealed that microfilaruria was still present and that living microfilariae of O. volvulus could be recovered from small urine samples obtained by high catherization of the ureters. Radiological changes consistent with chronic pyelitis were found in 4 out of 14 persons with microfilaruria who were examined by retrograde pyelography. The recovery of microfilariae in the urine was associated with the intensity of the infection, as determined by counts of microfilariae in skin snips and the number of onchocercomata. A systematic comparison for differences between onchocerciasis patients with and without microfilaruria revealed that the microfilaruria patients had a low weight: height ratio, deficient antibody responses in indirect haemagglutination tests with O. volvulus antigen, elevated serum aminotransferase levels, and reduced systolic blood pressure.     

Transmission in utero of microfilariae of Onchocerca volvulus.

Skin snips from newborn children and biopsy material from the umbilical cord and placenta of their mothers were examined. Microfilariae of O. volvulus were found in the skin of 2 out of 11 babies and in the umbilical cords of 2 out of 11 babies born to ten mothers infected with O. volvulus in Bawku Hospital, Ghana.     

Studies on onchocerciasis in the United Cameroon Republic. I. Comparison of populations with and without Onchocerca volvulus

The results of an onchocerciasis survey of total populations aged 5 years and over in 22 village groups in Cameroon rain-forest and savanna are reported. Using standardized techniques the same observers examined skin and eye lesions in 2,678 persons infected with O. volvulus, and compared them with lesions in 1,156 persons in whom the parasite was not detected.     

DNA immunisation with Onchocerca volvulus chitinase induces partial protection against challenge infection with L3 larvae in mice

The parasitic nematode, Onchocerca volvulus is a major cause of blindness and dermal pathology in tropical regions. A vaccine directed to infective larvae would provide a valuable control tool alongside the current methods of chemotherapy and vector control. Previously we have described the identification of a chitinase cDNA that is expressed in a stage specific manner by O. volvulus infective third stage (L3) larvae. To evaluate its host protective potential, the complete open reading frame was cloned into the eukaryotic expression plasmid pJW4303 and used to vaccinate mice by DNA immunisation with the Accell GeneGun. The survival of challenge infective larvae was monitored using implanted micropore chambers. In the first trial, mice immunised 3 times over 4 months with 1 μg O. volvulus chitinase DNA responded with modest antibody responses dominated by IgG2a and exhibited a 36% (p=0.189, NS) reduction in parasite survival compared with challenge controls. In the second trial, an increased dose of DNA (5 μg) and more frequent immunisations (5 times over 6 months) stimulated an IgG1 dominant response and a 53% reduction in parasite survival (p=0.042). Antibodies from the vaccinated mice reacted with the cuticle of post-infective L3 larvae, implying that this may be the site of immune attack following secretion of chitinase.     

Histochemical enzyme variation in Onchocerca volvulus microfilariae from rain-forest and Sudan-savanna areas of the Onchocerciasis Control Programme in West Africa

Histochemical staining methods for acid phosphatase were used to study the differences among microfilariae of various West African strains of Onchocerca volvulus in both forest and Sudan-savanna onchocerciasis zones. The results have shown statistically significant differences in the staining patterns of microfilarial populations in the two zones. In the rain-forest areas, where onchocerciasis is transmitted by Simulium yahense, S. sanctipauli, S. soubrense and S. squamosum, there were no significant differences of microfilarial staining patterns in patients, by age and sex, between the three Simulium—Onchocerca complexes studied. There was a close relationship between the “strain differences”, as revealed morphoenzymatically, and the clinical picture of the disease in both the forest and the Sudan-savanna zones. The present findings are in favour of the hypothesis that there are intrinsic differences in the strains of the parasite occurring in the two areas. The application of the histochemical means of parasite characterization appears to be a useful tool in differentiating strains of O. volvulus and could contribute towards a better understanding of the epidemiology of human onchocerciasis in different bioclimatic zones where the disease is endemic.     

Onchocerciasis in Kenya 9, 11 and 18 years after elimination of the vector

Elimination of the onchocerciasis vector Simulium neavei through larvicidal operations in focal areas of Kenya in 1946, 1953, and 1955 achieved complete interruption of transmission. Since no treatment was administered to the infected population, the areas provided an opportunity for studying the natural course of the infection in man in the absence of reinfection, with particular emphasis on its average duration and the effect of duration of exposure to the infection. In a follow-up survey conducted in 1964 in four focal areas, approximately 2000 people were examined parasitologically and clinically; slightly over half this group were also given a thorough ophthalmological examination. The results showed that, 11 years after interruption of transmission, live Onchocerca volvulus adults were present in nodules and microfilariae were present in the skin; after 18 years, however, microfilariae were no longer found in the skin. Assuming that in hyperendemic areas parasites are acquired until shortly before interruption of transmission, it can thus be postulated that O. volvulus worms lose their reproductive potentiality after 16 years or possibly earlier. A comparison of recent microfilarial rates with adjusted rates found in earlier surveys seems to indicate that the onchocercal infection, after interruption of transmission, follows a straight regression line, theoretically reaching zero after about 13-17 years.     

Artificial feeding of Simulium vectors of human and bovine onchoceriasis

A technique for feeding simuliids artificially in a feeding-tube containing blood covered by a membrane has been developed with a view to studying the development of Onchocerca volvulus of man and O. gutturosa of cattle in their respective vectors. A number of different kinds of membranes were tested, but skin from 2-day-old chicks was found to be most satisfactory. S. ornatum, the vector of O. gutturosa in England, fed readily on human and ox blood. The trials were carried out in a constant-temperature room with artificial lighting supplied by 60- or 100-watt clear blue-glass lamps illuminating the feeding-tube with 20 lumens per ft². Ultraviolet illumination was not satisfactory but light filtered through coloured screens gave good results in the middle of the spectrum. The flies fed well up to a temperature of 21°C (70°F) and a relative humidity of 65%-75%. The over-all feeding rate was 24.5%.     

Ocular onchocerciasis.

Well over 20 million people in the world are infected with Onchocerca volvulus and it is probable that 200 000-500 000 people are blind as a result of this infection, which is the most important cause of blindness in certain areas of Africa and Latin America.     

Inflammatory reactions in onchocerciasis: a report on current knowledge and recommendations for further study.

This report concerns the host''s reactions to the presence of the parasite both in the course of the natural disease and during drug treatment. The various stages of Onchocerca volvulus are discussed in terms of the type of tissue reaction seen. The discussion then turns to basic hypotheses concerning the etiology of these reactions, emphasis being placed on the fact that while pathological changes are considerable in some locations there is a remarkable lack of reaction in others. Some of the mechanisms possibly involved in this apparent absence of host response are discussed, including anti-complement factors, poor antigenicity, acquisition of host antigen, immune tolerance, and blocking antibodies. In any study of the inflammatory response it is recommended that critical evaluations be made of histological material, haematological studies, the definition of the antigenic nature of O. volvulus, characterization of immunological reactivity of patients, and the definition of the migratory pathways of the parasite.     

Seasonal variation in the microfilarial skin density of Onchocerca volvulus and in the biting activity of Simulium spp. in Guatemala

The density of Onchocerca volvulus microfilariae in the skin of 11 carriers was examined by carrying out a monthly examination in a single endemic area of Guatemalan onchocerciasis. The results obtained suggested the possibility of a seasonal variation in the microfilarial skin density. Almost all of the volunteers participating had shown a peak concentration of microfilariae in September, followed by August or October. The highest biting densities of Simulium ochraceum, the principal vector of onchocerciasis in Guatemala, occurred from September to the following January. It thus appears that the seasonal variation in the microfilarial density may correspond, to some extent, with the biting activity of the vector, S. ochraceum, and/or with climatic changes in the area.     

Robust gut associated vaccine-specific antibody-secreting cell responses are detected at the mucosal surface of Bangladeshi subjects after immunization with an oral killed bivalent V. cholerae O1/O139 whole cell cholera vaccine: Comparison with other mucosal and systemic responses

The emergence of V. cholerae O139 serogroup of V. cholerae capable of causing severe dehydrating cholera has over the decade led to efforts in formulation of vaccines to protect against this pathogen. Although the prevalence of diarrhea due to V. cholerae O139 has recorded a decrease, efforts on vaccine development continues to formulate an oral vaccine capable of stimulating the gut mucosal system. We have studied the mucosal immunogenicity in Bangladeshi adults to a killed whole cell (WC) bivalent cholera vaccine composed of V. cholerae O139 as well as V. cholerae O1 strains together with the recombinant cholera toxin B subunit (CTB) (WC-O1/O139/CTB) and compared the immune responses to that obtained with the licensed monovalent cholera vaccine, Dukoral (WC-O1/CTB). Direct estimation of the WC-O1/O139/CTB vaccine-specific mucosal responses were carried out using lymphocytes isolated from duodenal biopsies, intestinal lavage fluid and feces. The vaccine induced robust antibody-secreting cell responses in the duodenum specific to CTB as well as the O1 and O139 lipopolysaccharide (LPS). Magnitude of response was higher in the gut than in the circulation in all three antibody isotypes. The CTB and LPS-specific mucosal antibody responses were also seen in intestinal lavage fluid and fecal extracts. Vibriocidal antibody responses in plasma were observed to both the V. cholerae O1 and O139 serogroups (76% and 57% response rates, respectively). Plasma IgA and IgG responses to CTB and IgA responses to both O1 and O139 LPS were elevated. The immune responses were comparable to that seen to the monovalent WC-O1/CTB recipients in all components studied. Overall, the bivalent cholera vaccine induces strong mucosal responses and the addition of the O139 component does not interfere with the responses to the licensed vaccine Dukoral. This sets the ground for testing such vaccines in large field trials in Bangladesh and also demonstrates that addition of other vibrio components to the existing cholera vaccine does not alter the responses to the O1 vaccine components.     

Enhanced humoral response to influenza vaccine in aged mice with a novel adjuvant,rOv-ASP-1

Immunization is the best way to prevent seasonal epidemics and pandemics of influenza. There are two kinds of influenza vaccines available in the United States: an inactivated vaccine (TIV) and an attenuated vaccine; however, only TIV is approved for immunization of the elderly population. While the aged population has the highest rate of influenza vaccination, the protective efficacy is low as evidenced by elderly individuals having the highest mortality associated with influenza. Recently, we reported that an adjuvant derived from the helminth parasite Onchocerca volvulus, named O. volvulus activation-associated secreted protein-1 (Ov-ASP-1), can significantly enhance the protective efficacy of an inactivated vaccine (TIV) in young adult mice. In the current study, we examined whether this recombinant Ov-ASP-1 (rOv-ASP-1) can enhance the efficacy of TIV in aged mice as well. While primary immunization with TIV alone produced only a low level of influenza-specific antibodies (total IgG, IgG1, and IgG2c) in aged mice, the antibody levels were significantly increased after immunization with TIV + rOv-ASP-1. More importantly, the level of the total IgG in aged mice administered TIV + rOv-ASP-1 was comparable to that of young adult mice immunized with TIV alone. Co-administration of rOv-ASP-1 induced a low level of cross-reactive antibody and enhanced the protective efficacy of TIV in aged mice, reflected by significantly increased survival after challenge with a heterologous influenza virus. rOv-ASP-1 was also superior to the conventional adjuvant alum in inducing specific IgG after TIV immunization in aged mice, and in conferring protection after challenge. These results demonstrate that rOv-ASP-1 may serve as a potential adjuvant for influenza vaccine to improve the efficacy of protection in the elderly.     

The effects of drugs on Onchocerca volvulus. 1. Methods of assessment, population dynamics of the parasite and the effects of diethylcarbamazine

It is important to standardize quantitative methods for assessing the action of drugs on Onchocerca volvulus in man. There are certain disadvantages to making observations on adult worms removed from excised nodules in treated patients, but a great deal of information on drug action can be obtained by making a careful study of the concentrations of microfilariae in multiple weighed skin snips taken before treatment and at intervals after treatment.     

The effects of drugs on Onchocerca volvulus. 2. The antimonial preparations TWSb and MSbE

Antimonial preparations (Pentostam, Neostibosan, stibophen, and tartar emetic) have occasionally been used in the treatment of onchocerciasis without very promising results. The advent of the preparations TWSb (stibocaptate) and MSbE (Friedheim) of allegedly reduced toxicity made it desirable to test them against Onchocerca volvulus.     

Epidemiological studies on onchocerciasis by means of a new field technique.

A new membrane filter concentration technique for the detection and quantification of Onchocerca volvulus microfilariae in skin snips was compared for sensitivity and efficiency with a widely used “standard” technique. A field study was carried out in five villages in an onchocerciasis focus north-east of the town of Sokodé, Mô river valley, Togo. Use of the new technique resulted in a substantial rise in the observed prevalence and density of microfilariae.     

An evolutionary analysis of nitric oxide reductase gene norV in enterohemorrhagic Escherichia coli O157

A novel virulence gene, norV, that encodes nitric oxide (NO) reductase, was examined to investigate the emergence of enterohemorrhagic Escherichia coli (EHEC) O157 subgroup C clusters 2 and 3 from subgroup C cluster 1. Deletion of norV occurred at a point between cluster 1 and cluster 2 just after or at the same time that an stx2 bacteriophage, which retains Shiga toxin 2 gene, was inserted into wrbA, which encodes a novel multimeric flavodoxin-like protein, in EHEC O157. Sensitivity of NO to anaerobic growth was correlated with the deletion of norV in all EHEC O157 individuals tested. The C467A mutation of fimH, which encodes minor component of type 1 fimbriae, occurred within cluster 1, not as a transition from cluster 1 to cluster 2, indicating that there is a cluster 1 minority branch that leads to cluster 2. These data refine the evolutionary history of an emerging EHEC O157.     

The effects of drugs on Onchocerca volvulus. 4. Trials of melarsonyl potassium

The effects of the arsenical drug melarsonyl potassium on Onchocerca volvulus were investigated in patients in Cameroon infected with the Cameroon forest and Sudan savanna strains of the parasite. Two intramuscular dosage schedules were tested: the first comprised 4 consecutive daily doses of 200 mg repeated once after a 10-14 day interval, i.e., 2 (4×200 mg). The second was a single dose schedule at 7.1 mg/kg—10 mg/kg, with a maximum of 500 mg.     

The intake and transmissibility of Onchocerca volvulus microfilariae by Simulium damnosum fed on patients treated with Diethylcarbamazine, suramin or Mel W

The best hope for the control of transmission of Onchocerca volvulus over much of Africa lies in combining larvicidal measures directed against the Simulium vectors with chemotherapeutic measures designed to reduce the reservoir of transmissible microfilariae in man.