Biomedical subjects

G Cuny

Publications and source records attributed to G Cuny.

At least 19 recordsLinked to original sources

PubMed2006

Cyclical transmission of Trypanosoma brucei gambiense in Glossina palpalis gambiensis displays great differences among field isolates.

Six sets of teneral Glossina palpalis gambiensis (Diptera: Glossinidae) were fed on mice infected with six different isolates of Trypanosoma brucei gambiense (each mouse was infected with one of the isolates), previously isolated from patients in the sleeping sickness focus of Bonon, Côte d'Ivoire and in Makoua, Congo. All the tsetse flies were dissected 42 days post-infection and midgut and salivary glands were examined for trypanosomes by microscopical examination. No infection was observed with the reference stock whereas each of the five recently isolated trypanosome isolates was able to infect tsetse flies, with rates of infection varying between 9.7 and 18.2% depending on the isolate. Three isolates displayed only immature infections with 9.7, 17.3 and 18% of the flies showing trypanosomes in their midgut. One isolate gave both immature (12.1%) and mature infections (6.1%). Finally, the last isolate involved only mature infections in 9.7% of the Glossina species examined. These substantial differences in the cyclical transmission of T. b. gambiense in the same fly species could have important implications for the epidemiology of the transmission of Human African Trypanosomiasis.

Animals↗

PubMed2006

The tsetse fly Glossina palpalis palpalis is composed of several genetically differentiated small populations in the sleeping sickness focus of Bonon, Côte d'Ivoire.

Glossina palpalis is the main vector of human African trypanosomosis (HAT, or sleeping sickness) that dramatically affects human health in sub-Saharan Africa. Because of the implications of genetic structuring of vector populations for the design and efficacy of control campaigns, G. palpalis palpalis in the most active focus of sleeping sickness in Côte d'Ivoire was studied to determine whether this taxon is genetically structured. High and statistically significant levels of within population heterozygote deficiencies were found at each of the five microsatellite loci in two temporally separated samples. Neither null alleles, short allele dominance, nor trap locations could fully explain these deviations from random mating, but a clustering within each of the two samples into different genetic sub-populations (Wahlund effect) was strongly suggested. These different genetic groups, which could display differences in infection rates and trypanosome identity, were composed of small numbers of individuals that were captured together, leading to the observed Wahlund effect. Implications of this population structure on tsetse control are discussed.

Animals↗

PubMed2006

Aparasitemic serological suspects in Trypanosoma brucei gambiense human African trypanosomiasis: a potential human reservoir of parasites?

The serological and parasitological tests used for Trypanosoma brucei gambiense human African trypanosomiasis (HAT) diagnosis have low specificity and sensitivity, respectively, and in the field, control program teams are faced with subjects with positive serology but negative parasitology who remain untreated. The aim of this work was to explore, using PCR tool, the significance of these aparasitemic serological suspects. Since discordant PCR results have been observed earlier with different extraction methods, two DNA extraction methods were compared (the Chelex 100 resin and the DNeasy Tissue kit). The study was conducted on 604 blood samples: 574 from parasitologically confirmed patients, aparasitemic serological suspects and endemic controls collected in Côte d'Ivoire and 30 from healthy volunteers collected in France. No significant differences were observed between the PCR results obtained with the two extraction methods. Concerning PCR, problems of reproducibility and discordances with both serological and parasitological test results were observed, mainly for the aparasitemic serological suspects. In addition to previous results that pointed to the existence of non-virulent or non-pathogenic trypanosome strains and of individual susceptibility leading to long term seropositivity without detectable parasitaemia but positive PCR, the results of this study support the notion of a long lasting human reservoir that may contribute to the maintenance or periodic resurgences of HAT in endemic foci.

Agglutination Tests↗

PubMed2005

Wild fauna as a probable animal reservoir for Trypanosoma brucei gambiense in Cameroon.

In order to study the existence of a wild animal reservoir for Trypanosoma brucei gambiense in South Cameroon, blood was collected from wild animals in three human African trypanosomiasis foci and from a nonendemic control area. The 1142 wild animals sampled belonged to 36 different species pertaining to eight orders (407 primates, 347 artiodactyls, 265 rodents, 54 pangolins, 53 carnivores, 11 saurians and crocodilians, and five hyraxes). QBC and KIVI tests detected trypanosomes on 1.7% (13/762) and 18.4% (43/234) of animals examined, respectively. Using specific primers, T. brucei non-gambiense group 1 DNA was detected on 56 animals (4.9%). This infection rate was 5.3% in the endemic zone and 3.8% in the control zone. Of the 832 animals of the endemic zone, PCR revealed T. b. gambiense group 1 DNA in 18 (2.2%). These hosts included two rodents, two artiodactyls, two carnivores and two primates. T. b. gambiense group 1 was absent from animals from the nonendemic zone. A decrease in the prevalence of T. b. gambiense group 1 was observed in wild animals from the Bipindi sleeping sickness focus after a medical survey and vector control in this area. The epidemiological implications of these findings remain to be determined with further investigations.

Animals↗

PubMed2005

Spontaneous cure of domestic pigs experimentally infected by Trypanosoma brucei gambiense. Implications for the control of sleeping sickness.

The existence of a pig reservoir for human African trypanosomosis (HAT) due to Trypanosoma brucei gambiense complicates the fight against this disease. This study, reports results obtained from pigs, which were inoculated with the blood of a person, suffering from HAT in Cameroon. The pigs were reared and kept in the shelter from all contact with Glossina, and monitored for 188 days. The seroconversion was checked by agglutination assays for trypanosomosis (CATT 1.3 and LATEX/T.b.gambiense). The parasitemia was measured by quantitative buffy coat method (QBC) and by polymerase chain reaction method (PCR). In addition, growth was recorded as well as blood counting and blood formulas. The results showed that the pigs were trypanotolerant and cure themselves in less than 6 months. It is concluded that sterilisation of this reservoir could be achieved by tsetse-control measures in 1 year. It confirms the strategy to complement screening and treatment of HAT with tsetse fly control measures.

Animals↗

PubMed2005

Trypanosoma brucei s.l.: characterisation of stocks from Central Africa by PCR analysis of mobile genetic elements.

To better understand the epidemiology of sleeping sickness in the Central African sub-region, notably the heterogeneity of Human African Trypanosomiasis (HAT) foci, the mobile genetic element PCR (MGE-PCR) technique was used to genotype Trypanosoma brucei s.l. (T. brucei s.l.) isolates from this sub-region. Using a single primer REV B, which detects positional variation of the mobile genetic element RIME, via amplification of flanking regions, MGE-PCR revealed a micro genetic variability between Trypanosoma brucei gambiense (T. b. gambiense) isolates from Central Africa. The technique also revealed the presence of several T. b. gambiense genotypes and allowed the identification of minor and major ubiquitous genotypes in HAT foci. The presence of several T. b. gambiense genotypes in HAT foci may explain the persistence and the resurgence phenomena of the disease and also the epidemic and the endemic status of some Central African sleeping sickness foci. The MGE-PCR technique represents a simple, rapid, and specific method to differentiate Central African T. brucei s.l. isolates.

Africa, Central↗

PubMed2005

[Human African trypanosomiasis: urban transmission in the focus of Bonon (Côte d'Ivoire)].

Human African trypanosomiasis (HAT) is a vector-borne parasitic disease which has often been considered a rural disease. Population increases in African countries have entailed the spread of urban centres, creating favourable conditions for the appearance of new epidemiological conditions. In Cote d'Ivoire, HAT transmission has been described in the surroundings of towns such as Daloa or Sinfra. In the focus of Bonon, located in central-western Cote d'Ivoire, a medical survey detected 96 patients. The sites visited by the patients every day were geo-referenced and the routes between them recorded. In parallel, an entomological survey of the patients' daily locations enabled the collection of data on the vector. In Bonon, we observed urban cases and tsetse (Glossina palpalis) feeding on men. Trypanosoma brucei gambiense was identified in both man and vector; thus all conditions for possible intra-urban trypanosomosis transmission were met. The consequences of this are discussed regarding the problem of diffusion of the disease.

Animals↗

PubMed2004

A study of host preference in tsetse flies using a modified heteroduplex PCR-based method.

A study of host preference in tsetse flies using a modified heteroduplex PCR-based method is described. Domestic and wild animal blood samples were collected to extract the corresponding reference DNAs. In Campo (south Cameroon), tsetse flies (mainly Glossina palpalis palpalis) were trapped and 41 bloodmeals were collected. All reference DNAs and 37 bloodmeal DNAs (90.7%) were successfully amplified and hybridised. Twelve bloodmeals (32.4%) were of human origin, 13 (35.4%) were from Sitatunga (Tragelaphus spekei) (an antelope) while 12 (32.4%) were not identified using our set of reference DNAs. The results confirmed the occurrence of frequent contacts between wild animals and this population of tsetse flies.

Animals↗

PubMed2004

Mixed infections of trypanosomes in tsetse and pigs and their epidemiological significance in a sleeping sickness focus of Côte d'Ivoire.

In a sleeping sickness focus of Côte d'Ivoire, trypanosomes were characterized in humans, pigs and tsetse using various techniques. Out of 74 patients, all the 43 stocks isolated by KIVI (Kit for In Vitro Isolation) appeared to belong to only one zymodeme of Trypanosoma brucei gambiense group 1 (the major zymodeme Z3). The only stock isolated on rodents belonged to a different, new, zymodeme (Z50), of T. b. gambiense group 1. From 18 pigs sampled in the same locations as the patients, PCR showed a high proportion of mixed infections of T. brucei s. l. and T. congolense riverine-forest. Zymodemes of T. brucei s. l. from these pigs were different from those found in humans. From a total of 16 260 captured tsetse (Glossina palpalis palpalis), 1701 were dissected and 28% were found to be infected by trypanosomes. The most prevalent trypanosome was T. congolense riverine-forest type, followed by T. vivax, T. bruceis. l. and T. congolense savannah type, this latter being associated to the forest type of T. congolense in most cases. Mixed infections by 2 or 3 of these trypanosomes were also found. Use of a microsatellite marker allowed us to distinguish T. b. gambiense group 1 in some of the mature infections in tsetse. Differences in infection rates and in trypanosome genotypes according to the host might indicate that the pig may not be an active animal reservoir for humans in this focus.

Animals↗

PubMed2004

Inhibition of the DNA amplification of trypanosomes present in tsetse flies midguts: implications for the identification of trypanosome species in wild tsetse flies.

The present study was carried out in order to investigate if there was really a failure of PCR in identifying parasitologically positive tsetse flies in the field. Tsetse flies (Glossina palpalis gambiensis and Glossina morsitans morsitans) were therefore experimentally infected with two different species of Trypanosoma (Trypanosoma brucei gambiense or Trypanosoma congolense). A total of 152 tsetse flies were dissected, and organs of each fly (midgut, proboscis or salivary glands) were examined. The positive organs were then analysed using PCR. Results showed that, regardless of the trypanosome species, PCR failed to amplify 40% of the parasitologically positive midguts. This failure, which does not occur with diluted samples, is likely to be caused by an inhibition of the amplification reaction. This finding has important implications for the detection and the identification of trypanosome species in wild tsetse flies.

Animals↗

PubMed2004

Monitoring the susceptibility of Glossina palpalis gambiensis and G. morsitans morsitans to experimental infection with savannah-type Trypanosoma congolense, using the polymerase chain reaction.

Teneral Glossina palpalis gambiensis and G. morsitans morsitans (Diptera: Glossinidae) were fed on mice infected with savannah-type Trypanosoma (Nannomonas) congolense. The infection was monitored by checking the post-feeding diuresis fluid (midgut infection) and saliva (mature infection) of individual flies for parasites, at different times post-infection, using microscopical examination and a PCR-based assay. The results indicated that both tsetse species supported established midgut infections by 10 days post-infection and that maturation occurred after 24 days in G. m. morsitans. Although, for both diuresis fluid and saliva, the results of the microscopy showed good concordance with those of the PCR, the PCR identified more positive samples. Monitoring allowed determination of the status of the infection in individual flies, which was confirmed, 48 days post-infection, by the microscopical examination of the midguts and probosces dissected out of the flies and by the PCR-based amplification of any trypanosome DNA in these organs. Again, in terms of the detection of trypanosomes in the dissected organs, there was good concordance between the results of the PCR and those of the microscopy, although PCR revealed many more mature infections than did microscopical examination, particularly in the G. p. gambiensis investigated. There was a higher prevalence of immature infection in G. p. gambiensis than in G. m. morsitans (P<0.05) but the inter-specific differences seen in the prevalences of any infection and of mature infection were not statistically significant. The intrinsic vectorial capacity for T. congolense of both tsetse species therefore appeared quite similar, although the true vectorial competence of G. p. gambiensis remains to be determined.

Animals↗

PubMed2004

Characterization of Trypanosoma brucei s.l. infecting asymptomatic sleeping-sickness patients in Côte d'Ivoire: a new genetic group?

Six villagers in the Sinfra focus of sleeping sickness in Côte d'Ivoire who in 1995 were asymptomatic and refusing treatment, despite then being serologically and parasitologically positive for trypanosomes, were followed-up, while still refusing treatment, until 2002. In 2002, five of the six cases remained serologically positive but no trypanosomes could be found in any of them by use of the classical parasitological methods. A PCR-based assay, however, revealed that all six had the DNA of Trypanosoma brucei s.l. in their blood, so confirming the low sensitivity of the classical parasitological tests. The analysis of satellite, minisatellite and microsatellite markers indicated that, in 2002, all six cases were infected with a 'new' distinct genetic group of T. brucei s.l. and four were co-infected with T. b. gambiense group 1. The epidemiological consequences of such co-infections are discussed. The 'new' group of T. brucei had a molecular pattern that differed from those of the classical T. b. gambiense group 1 and the 'bouaflé' group.

Animals↗

PubMed2003

Monitoring the developmental status of Trypanosoma brucei gambiense in the tsetse fly by means of PCR analysis of anal and saliva drops.

Teneral Glossina palpalis gambiensis (Diptera: Glossinidae) were infected with a culture of procyclic forms of Trypanosoma brucei gambiense using a single-bloodmeal membrane feeding technique. The infection was monitored by analysing the saliva (mature infection) and anal drop (midgut infection) of each fly at different post-infection times both by microscopic observation and polymerase chain reaction (PCR). Amplification revealed many more positive anal drops than microscopy. The monitoring showed that the installation of T. b. gambiense in Glossina took place at least 11 days after the infection and that maturation occurred after 29 days. It also reflected precisely the parasitic status of each tsetse fly as determined by the dissection, microscopic examination and PCR amplification of the midguts and salivary glands 47 days post-infection. Twice as many tsetse flies with mature salivary glands infection were revealed by PCR than by microscopic examination, but the two techniques gave exactly the same results regarding the proportion of flies with midgut infection. This study also demonstrated the ability of natural non-infective procyclic forms of T. b. gambiense, to colonise the midgut and subsequently establish in the salivary glands of G. p. gambiensis.

Anal Canal↗

PubMed2003

Stage determination and therapeutic decision in human African trypanosomiasis: value of polymerase chain reaction and immunoglobulin M quantification on the cerebrospinal fluid of sleeping sickness patients in Côte d'Ivoire.

In human African trypanosomiasis (HAT), two disease stages are defined: the first, or haemo-lymphatic stage, and the second, or meningo-encephalitic stage. Stage determination forms the basis of therapeutic decision and is of prime importance, as the drug used to cure second-stage patients has considerable side-effects. However, the tests currently used for stage determination have low sensitivity or specificity. Two new tests for stage determination in the cerebrospinal fluid (CSF) were evaluated on 73 patients diagnosed with HAT in Côte d'Ivoire. The polymerase chain reaction (PCR) detecting trypanosome DNA (PCR/CSF) is an indirect test for trypanosome detection whereas the latex agglutination test detecting immunoglobulin M (LATEX/IgM) is an indicator for neuro-inflammation. Both tests were compared with classically used tests, double centrifugation and white blood cell count of the CSF. PCR/CSF appeared to be the most sensitive test (96%), and may be of use to improve stage determination. However, its value for therapeutic decision appears limited, as patients whose CSF was positive with PCR were successfully treated with pentamidine. This result confirms those of previous works that showed that some patients with trypanosomes in the CSF could be treated successfully with pentamidine. LATEX/IgM, which depending on the cut-off, showed lower sensitivity of 76% and 88%, but higher specificity of 83% and 71% for LATEX/IgM 16 and LATEX/IgM 8 respectively, appears more appropriate for therapeutic decision making.

Animals↗

PubMed2002

Microsatellite markers for population genetic studies in Aedes aegypti (Diptera: Culicidae) from Côte d'Ivoire: evidence for a microgeographic genetic differentiation of mosquitoes from Bouaké.

In West Africa, Aedes aegypti (Diptera: Culicidae) (Linnaeus, C., 1762. Zweyter Theil, enhalt Beschreibungen veschiedener wichtiger Naturalien. In: Hasselquist, F. (Ed.), Reise nach Palastina in den Jahren von 1749 bis 1752, Rostock, Germany, pp. 267-606) represents the principal vector of yellow fever. This study reports the use of microsatellite markers to characterise various A. aegypti populations from Côte d'Ivoire according to a north-south transect, and to perform a temporal genetic survey of the mosquitoes. Three microsatellite loci were used to analyse individuals from four different places: Kabolo, Bouaké, and two different districts of Abidjan. We found that the four populations are genetically distinct except the two Abidjan populations. In the Bouaké population, the coexistence of two cryptic species, not morphologically distinguishable, seems to account for the extensive heterozygote deficiency observed. Comparison of mosquitoes from Bouaké 1 year apart indicated that a dramatic change occurred in the structuring of this population over time. Taken together these results indicate that microsatellite markers could be useful for identifying various populations of A. aegypti on a microgeographic scale and to assess for temporal variation within mosquito populations.

Aedes↗

PubMed2002

Genetic characterization of Trypanosoma brucei gambiense and clinical evolution of human African trypanosomiasis in Côte d'Ivoire.

Human African trypanosomiasis is a parasitic infection caused by protozoa belonging to Trypanosoma brucei subspecies. The clinical evolution of this disease is complex and might be because of the parasite itself, as genetic diversity has been observed in T. brucei ssp. We investigated the relationship between the genetic diversity of trypanosomes and the diversity of clinical patterns in Côte d'Ivoire. We studied clinical sleeping sickness cases, and genetically analysed the trypanosomes isolated from these patients. An important genetic monomorphism among stocks isolated in Côte d'Ivoire was observed by using various markers: isoenzymes electrophoresis, random amplified polymorphism DNA and PCR of microsatellite sequences. At the same time, the diversity of clinical patterns and evolutions was confirmed by clinical analysis. The existence of an individual susceptibility to disease (human trypanotolerance) should be taken into account even if our genetic conclusions might be distorted because the isolation success rates were particularly poor. In fact, we observed that the isolation success rate varied significantly depending both on the focus of origin (P=0.0002) and on the ethnic group (P=0.0317) of the patient. Further investigations are required in order to study a possible selective impact of the use of the kit for in vitro isolation of trypanosomes as an isolation technique.

Animals↗

PubMed2001

New molecular marker for Trypanosoma (Duttonella) vivax identification.

Trypanosoma vivax is a widespread hemoparasite in tropical areas and is pathogenic to ruminant domestic livestock as well as wild ruminants. The accurate identification of parasites in both hosts and vectors is crucial for epidemiological studies and disease control programs. We describe here the development of molecular markers specific for T. vivax identification. These markers were used to identify mouthpart infections in field-collected tsetse flies from Cameroon. The markers target the genomic sequence of a species-specific antigen from the bloodstream stages. No cross amplification with other trypanosome species was observed, which makes the markers a reliable tool to detect T. vivax infections, both in hosts and vectors. The PCR-amplified sequence contains a (CA)(n) microsatellite repeat for which 11 different alleles were identified. This microsatellite, which showed high polymorphism, provides a suitable marker for population genetic studies.

Animals↗

PubMed2001

A preliminary study of the population genetics of Aedes aegypti (Diptera: Culicidae) from Mexico using microsatellite and AFLP markers.

Dengue fever recently reemerged in the Americas. Because vaccines are still under development, dengue prevention depends entirely on vector control. Since Aedes aegypti (Linnaeus, 1762) is the principal vector of this arbovirus, knowledge of the genetic structure of the insect is therefore required to maintain effective vector control strategies and to estimate levels of gene flow from which movement can be inferred. This preliminary study uses microsatellite and amplified fragment length polymorphism (AFLP) markers, to provide insights into genetic diversity of A. aegypti populations from different districts of two towns, located in the north-west of Mexico, Hermosillo and Guaymas. Although the microsatellites used were found to display limited polymorphism, they allowed discrimination between mosquitoes from the northern and the southern districts of Hermosillo. Using AFLP markers, clustering of individuals from the same town and from the same district was observed. Data from microsatellite and AFLP markers analysis both suggest that reinvasion of A. aegypti probably occurs from Guaymas to Hermosillo.

Aedes↗