Monitoring the pleomorphism of Trypanosoma brucei gambiense isolates in mouse: impact on its transmissibility to Glossina palpalis gambiensis.

Substantial differences have been observed between the cyclical transmission of three Trypanosoma brucei gambiense field isolates in Glossina palpalis gambiensis (Ravel et al., 2006). Differences in the pleomorphism of these isolates in rodent used to provide the infective feed to Glossina, could explain such results, since stumpy forms are preadapted for differentiation to procyclic forms when taken up in a tsetse bloodmeal. To assess this possibility, mice were immunosuppressed and inoculated intraperitoneally with the three isolates (six mice for each trypanosome isolate); then parasitaemia and pleomorphism were determined daily for each mouse. The three T. b. gambiense isolates induced different infection patterns in mouse. The parasitaemia peak was rapidly reached for all the isolates and maintained until mice death for two isolates, while the third isolate rapidly showed a falling phase followed by a second parasitaemia plateau. The proportion of the stumpy forms varied from 15% to 70% over the duration of the experiment and according to the isolate. One isolate, which displayed the highest proportion of stumpy forms and reached the stumpy peak at the onset of the falling phase of parasitaemia, was used to study the relationship between the proportion of stumpy forms and transmissibility to tsetse fly. The results indicated that the transmissibility of trypanosomes was not correlated to the proportion of non-dividing stumpy forms.

First outbreak of Trypanosoma evansi in camels in metropolitan France.

The first outbreak of trypanosomosis caused by Trypanosoma evansi in camels in France was reported on a farm in the Aveyron Department. Five camels were imported from the Canary Islands to the farm in early July 2006, and trypanosomes were observed on a stained blood smear from one of them, which died in October. On further investigations, trypanosomes were observed in the blood of five camels, three of them indigenous to the farm and two that had been imported. On the basis of microscopical examination (morphological criteria and measurements) and serological results based on the card agglutination T evansi test and PCR typing, the parasites were identified as T evansi. After treatment with melarsomine, the infected camels rapidly became negative by parasitological tests and were negative two to four months later by serological tests. The parasite was probably transmitted by tabanids and Stomoxys calcitrans, which were abundant in July to September 2006. No parasites were observed in other animals on the farm or on neighbouring farms, but some of the sheep on these farms were positive by PCR or serology.

Trypanosoma brucei brucei induces alteration in the head proteome of the tsetse fly vector Glossina palpalis gambiensis.

Parasitic manipulations of host behaviour are known from a wide range of host-parasite associations. However, the understanding of these phenomena is far from complete and detailed investigation of their proximate causes is needed. Many studies report behavioural modifications, such as altered feeding rates in tsetse fly (Glossina) infected with the mature transmissible stage (i.e. metacyclic) of the trypanosomes. Here, bidimensional (2D) gel electrophoresis and mass spectrometry were employed to analyse and compare the head proteome between four Glossina palpalis gambiensis categories (uninfected, refractory, mature infection, immature infection). Twenty-four protein spots specifically present or absent in the head of metacyclic-infected flies were observed. These protein spots were subsequently identified and functionally classified as glycolitic, neurotransmiter synthesis, signalling, molecular chaperone and transcriptional regulation proteins. Our results indicate altered energy metabolism in the head of metacyclic-infected tsetse flies. Some of the proteins identified, such as casein kinase 2 and jun kinase have previously been shown to play critical roles in apoptosis in insect neurones. In addition, we found two pyridoxal-dependent decarboxylases (dopa decarboxylase and alpha methyldopa hypersensitive protein), suggesting a modification of serotonin and/or dopamine in the brain of metacyclic-infected tsetse flies. Our data pave the way for future investigation of the alteration of the glossina central nervous system during infection by trypanosomes.

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.

Predominance of subtype A and G HIV type 1 in Nigeria, with geographical differences in their distribution.

The purpose of this study was to generate data on the relative prevalences of the HIV-1 subtypes circulating in Nigeria. A total of 252 HIV-1-positive samples collected during an epidemiologic survey conducted in April 1996 were genetically characterized by HMA (heteroduplex mobility assay) and/or sequencing. Samples were collected in Lagos, Calabar, Kano, and Maiduguri. Overall, the predominant env subtypes were A (61.3%) and G (37.5%). Subtype A is more prevalent in the south (p < 0.001), about 70% in Lagos and Calabar, whereas a quarter of the samples was classified as subtype G in these states. In contrast, subtype G is predominant in the north ( < 0.001), representing 58% of the samples in Kano. In the northeastern region, Maiduguri, almost similar proportions of subtype A and G were seen, 49 and 47.4%, respectively. A total of 37 samples was also sequenced in the p24 region from the gag gene; 13 (35%) had discordant subtype designations between env and gag. The majority of the gag (12 of 17) and env (14 of 22) subtype A sequences clustered with the A/G-IBNG strain. Within subtype G, three different subclusters were seen among the envelope sequences. These different subclusters are observed among samples obtained from asymptomatic individuals and AIDS patients from the four Nigerian states studied. In conclusion, we observed a limited number of HIV-1 subtypes circulating in Nigeria, with subtypes A and G being the major env subtypes responsible for the HIV-1 epidemic. Nevertheless, the high rate of recombinant viruses (A/G) and the different A/G recombinant structures indicate a complex pattern of HIV-1 viruses circulating in this country.

Evidence for differences in MT2 cell tropism according to genetic subtypes of HIV-1: syncytium-inducing variants seem rare among subtype C HIV-1 viruses.

Non-syncytium-inducing (NSI) variants seem to be more readily transmitted than syncytium-inducing (SI) variants, and the switch from NSI to SI during HIV-1 infection seems to be a key determinant to the evolution of AIDS. We investigated eventual differences in the SI capacity on MT-2 cells according to genetic subtypes of HIV-1 and correlated this observations with CD4 counts and duration of HIV infection. In total, 86 patients, most with known date of HIV contamination and infected with different genetic subtypes, have been studied: 11 subtype A, 46 subtype B, 22 subtype C, and 7 subtype E. Multivariate analysis used a Cox's proportional hazards regression. The number and percentage of patients infected with an SI strain were as follows: 3 of 11 (27%) for subtype A, 15 of 46 (33%) for subtype B, 0 of 22 (0%) for subtype C, and 5 of 7 (71%) for subtype E. After adjustment for time after seroconversion and CD4 counts, significantly fewer SI variants were observed in patients infected with subtype C (p < .002) and it was found that subjects infected with subtype E had a higher risk of being infected with an SI strain (rate ratio [RR] = 12.39%; 95% confidence interval [CI] 1.55-98.67; p < .001). Most of the subtype E-infected patients from our study switched from an NSI to SI phenotype early after seroconversion (<4 years). To predict the in vitro presence of SI variants, we scanned V3-loop sequences for mutations at positions 11 and/or 25. Overall, 54 of 55 (98.2%) NSI strains in vitro were predicted NSI, and only 4 of 12 (33.3%) of SI viruses were predicted SI. For patients in whom a switch from an NSI to an SI virus was observed, the SI phenotype could be detected earlier in vitro than by the corresponding V3-loop sequence. No SI strains were observed among patients infected with subtype C; however, longer follow-up is needed to see whether the appearance of SI variants in subtype E or the absence of SI variants in subtype C-infected patients is also associated respectively with a faster or slower progression to AIDS as described for subtype B.

Seroprevalence of HIV-1, HIV-2, and HIV-1 group O in Nigeria: evidence for a growing increase of HIV infection.

To determine current data on HIV infection and to further confirm the presence of HIV-1 group O infection in Nigeria, 2300 samples from five states were tested for the presence of HIV antibody. A convenience sampling was obtained from pregnant women, tuberculosis (TB) patients, commercial sex workers (CSWs), blood donors, patients with sexually transmitted diseases (STDs), patients with skin diseases, male clients of CSWs, outpatients suspected to have AIDS, truck drivers, and community dwellers. With the exception of pregnant women, the HIV prevalences in all these groups were high: 60.6% in CSWs, 16.2% in TB patients, 7.7% in blood donors in some states, and 16% in the rural area of Kano State. Male clients of CSWs, truck drivers, and STD patients had prevalences of 7.8%, 8.6%, and 21.2%, respectively. Regional differences in relation to HIV prevalences were observed; HIV-2 and most of the HIV-1/2 infections were found in the southern states of Nigeria. Higher HIV prevalences were observed in the north-northeast in pregnant women, TB patients, and CSWs, but for blood donors, higher rates were seen in the southeast-southwest. One asymptomatic 50-year-old woman, a community dweller in Kano, was identified to be HIV-1 group O-positive. Compared with data from national surveillance studies in 1991/1992 and 1993/1994, a substantial increase in HIV infection was observed. Our results show a growing incidence of HIV infection in Nigeria and suggest the presence of a rural HIV epidemic. The identification of HIV-1 group O in Kano shows that this virus strain is geographically widespread in Nigeria.

PIP: To obtain current data on HIV infection in Nigeria by population group, a seroanalysis of 2300 samples from 5 states (Lagos, Cross River, Borno, Kano, and Jugawa) was conducted during March-May 1996. The sample included commercial sex workers, pregnant women, tuberculosis patients, blood donors, patients with sexually transmitted diseases (STDs), patients with skin diseases, male clients of commercial sex workers, outpatients suspected to have AIDS, truck drivers, and community residents. Overall HIV prevalence was 40.7%. With the exception of pregnant women (1.7%), HIV prevalence was high in all subgroups: 60.6% in commercial sex workers, 21.2% in STD patients, 16.2% in tuberculosis patients, and 16.0% in rural areas of Kano state. The majority of HIV-positive women were 21-30 years of age, while HIV-infected men were primarily over 40 years of age. Compared with data from national seroprevalence studies conducted during 1991-92 and 1993-94, this study confirms a substantial recent increase in HIV infection in Nigeria. Of the 330 antibody-positive specimens, HIV-1 was the predominant infection in 315; there were 3 cases of HIV-2 and 12 cases involving dual HIV-1/2 infection. Only 1 serum sample was positive for HIV-1 group O antibodies. The high HIV prevalence detected among commercial sex workers indicates the potential for rapid diffusion of HIV to the general population.

Presence of multiple non-B subtypes and divergent subtype B strains of HIV-1 in individuals infected after overseas deployment.

OBJECTIVE: To identify the genetic subtypes and characteristics of HIV-1 strains from individuals infected after overseas deployment. PATIENTS AND METHODS: Sixty-one HIV-1-positive individuals detected between 1986 and 1995 in the French army were included in the study. For each patient, the year and country of HIV infection are known. Genetic subtypes of HIV-1 were determined using the heteroduplex mobility assay (HMA) using ED5/ED12 as outer and ES7/ES8 as inner primers. Strains were further characterized by sequencing and phylogenetic analysis of the C2-V3 region. The amino-acid sequences corresponding to the V3 region were aligned on the basis of the subtyping results and were then compared to the consensus V3 sequences of the corresponding subtypes. RESULTS: Among the 61 patients studied, nine became infected in France, and 52 were HIV-negative before overseas deployment but HIV-positive at their return. The majority (n = 43) deployed in Africa and a limited number of patients deployed in Asia (Cambodia, n = 5) or South America (guyana, n = 4). The nine individuals who were not deployed overseas were all infected with subtype B strains. The majority of the other patients were infected with non-B strains; eight subtype A, 20 subtype B, 16 subtype C, one subtype D, six subtype E and one subtype F. Five of the six subtype E strains were contracted in Cambodia and one in Djibouti, and all subtype C strains were from Djibouti. Phylogenetic analysis revealed a large diversity among the different strains introduced into France. Analysis of the amino-acid sequences of the V3 loop revealed the introduction of uncommon V3-loop patterns. CONCLUSION: In the group of HIV-1-infected individuals that we studied and who were deployed overseas, 63.4% were infected with non-B strains. In addition, the subtype A, B and C viruses in this population were very heterogeneous. Due to the routine occurrence of international travel and deployment, the predominance of subtype B HIV-1 viruses may change in European countries. However, the possible implications on the dynamics of the HIV-1 epidemic needs further follow-up.