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HBV genotyping was performed by sequencing and phylogenetic analyses of the surface (S) and core (C) fragments. Briefly, HBV DNA was first extracted from 400 µL of plasma and eluted in 100 µL of pure water, using the QIAamp Viral DNA Mini Kit (QIAGEN, Courtaboeuf, France) followed by semi-nested PCR amplification of the S (930 bp) and C (1010 bp) gene fragments using MP Taq Core Kits 25 (MP Biomedical Diagnostic, Europe). The S fragment amplification was performed as described elsewhere. (Hu X, et al., 2000 [PMID:10677515] ; Makuwa M et al., 2006 [PMID: 16847965]; Olinger CM et al.,2006 [PMID: 16603517]) The first round was performed using primers sets 58P (5’-CCT GCT GGT GGC TCC AGT TC-3’) and 979 (5’-ATT GGA AAG TAT GTC AAA GAA TTG TGG GTC TTT TG-3’). The 50 µL final reaction mixture contained 31.4 µL of RNase DNase Free water, 5 µL of buffer 10X with MgCl2 (25 mM), 0.4 µL of dNTPs (25 mM), 1.5 µL of each primer (10 µM), 0.2 µL of Taq polymerase (5 U/µL) and 10 µL of extracted DNA. The second round PCR used the 58P/Mc2r (5’-TGGAAGTTGGGGATCATTGCC-3’) primer on a 50 µL final reaction mixture containing 36.4 µL of RNase DNase Free water, 5 µL of buffer 10X with MgCl2 (25 mM), 0.4 µL of dNTPs (25 mM), 1.5 µL of each primer (10 µM), 0.2 µL of Taq polymerase (5 U/µL), and 5 µL of first round PCR product. The PCR program was the same for the first and second round PCRs including denaturation at 94°C for 5 min followed by 40 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 30 s and elongation at 72°C for 1 min, followed by final elongation at 72°C for 5 min. For the C fragment amplification, the couplet of primers BCP1F (5’-GCA TGG AGA CCA CCG TGA AC-3’) / 2853N (5’-TCA CCA TAT TCT TGG GAA CA-3’) was used for the first round and the couplet BCP2F(5’-CAT AAG AGG ACT CTT GGA CT-3’) / 2853N for the second round. The amplification conditions were the same as for the S fragment. The PCR products were purified using QIAquick PCR Purification Kit (Qiagen, Courtaboeuf, France) and submitted for sequencing at Macrogen Inc (Meibergdreef, Netherlands).

HBV DNA Real Time Quantification according to ARNS 12187 project Using the Arrow extractor (NorDiag, Biotrin International, Ireland), DNA was extracted from 240 μl of plasma, pre-treated with10 μl of proteinase K, using the Arrow Viral NA extraction kit, according to the manufacturer’s instructions during 45 min. Template DNA was eluted into 60 μl of kit elution buffer. Besides clinical samples, quantification standard (Acrometrix HBV Panel, Acrometrix, Menica, CA, USA) was also extracted for each run with the same protocol and1:10 diluted (from 50,000,000IU/mL to50IU/mL). For amplification, we used a primers/probe set designed under the auspices of “Agence Nationale de Recherches sur le SIDA et les hépatites virales” (ANRS 12187 project) and targeting a conserved region in the HBVS gene (nucleotide (nt) positions, 379–426). All runs were performed in a 50-μl volume containing DNA extract (10 μl), Master Mix (Platinum UGD, USA) (25 μl), pure water (HyClone Pure Water, Thermo Fisher Scientific, Waltham, MAUSA)(12.5 μl), forward primHBV1 (5’-GTGTCTGCGGCGTT TTATCA-3’) and reverse primHBV2 (5’-AGGCATAGCAGCAGGAT GAA-3’) primers at 10 μM (1μl each) and probe (5’FAM-TGCGGCGTTTTATCAT-MGB3’) at 5μM (0.5 μl). Each reaction consisted of: 2min at50°C and 10min at95°C; followed by 50 cycles of 15sec at 95°C and 1 min at 60°C each. The lower limit of quantification (LLOQ) of our technique was 100IU/mL and the lower limit of detection (LLOD) was 50IU/mL.

L’acces aux antiretroviraux a permis de prolonger l’esperance de vie des patients vivant avec le VIH. Toutefois, les co-infections VIH-hepatites virales, dans les pays endemiques pour ces infections, constitue un probleme pour une bonne prise en charge. Afin de connaitre l’ampleur de telles co-infections au Gabon, nous avons recherche, par technique ELISA, les marqueurs d’infection par les virus des hepatites B, C, et E chez 762 adultes infectes par le VIH-1. La quantification de l’ADN plasmatique du VHB (CV VHB) a ete effectuee par PCR en temps reel. Les amplifications des ARN VHC et VHE ont ete effectuees par RT-PCR et PCR nichees. La seroprevalence de l’infection a hepatite B chronique etait de 9,3 %. Parmi les patients avec des profils AcHBc isoles, la prevalence de l’infection a hepatite B occulte etait de 26,7 %, soit une prevalence reelle (CV-VHB detectables) de 17,3 % sur la population totale. La seroprevalence VHC etait de 8,8 %. Parmi les 67 patients seropositifs, 76,1 % avaient des profils replicatifs (ARN VHC detectables), soit une prevalence globale de 6,7 % sur la population totale. Pour l’hepatite E la seroprevalence etait de 3,5 %. Aucune infection chronique VHE n’a ete retrouvee. En conclusion, cette etude met en evidence un fort taux de co-infections VIH-Hepatites B, C et E au Gabon. Par ailleurs, nous montrons l’interet de rechercher les infections chroniques (profils replicatifs) chez les patients infectes par le VIH au Gabon. La mise en place de plateaux techniques pour ce type de recherche, accessible aux pays a revenus intermediaires s’impose.

Simian immunodeficiency virus (SIV) infection in African nonhuman primate (NHP) natural hosts is usually nonpathogenic, despite high levels of virus replication. We have previously shown that chronic SIV infection in sooty mangabeys (SMs) and African green monkeys (AGMs) is associated with low levels of immune activation and bystander T cell apoptosis. To compare these features with those observed in another natural host, the mandrill (MND), we conducted a cross-sectional survey of the 23 SIV-infected and 25 uninfected MNDs from the only semifree colony of mandrills available worldwide. Viral loads (VLs) were determined and phenotypic and functional analysis of peripheral blood- and lymph node-derived lymphocytes was performed. We found that mandrills chronically infected with SIVmnd-1 or SIVmnd-2 have similar levels of viral replication, and we observed a trend toward lower CD4 + T cell counts in chronically SIVmnd-2-infected MNDs than SIVmnd-1-infected MNDs. No correlation between CD4 + T cell counts and VLs in SIV-infected MNDs could be established. Of note, the levels of T cell activation, proliferation, and apoptosis were comparable between SIVmnd-1- and SIVmnd-2-infected MNDs and to those observed in uninfected animals, with the only exception being an increase in tumor necrosis factor alpha-producing CD8 + T cells in SIVmnd-2-infected MNDs. Overall, these findings recapitulate previous observations in SIV-infected SMs and AGMs and lend further evidence to the hypothesis that low levels of immune activation protect natural SIV hosts from disease progression.

Terra Nova, 23, 162–170, 2011 Abstract Most fault-related pseudotachylytes in the Val Sesia area, Ivrea zone, NW Italian Alps, occur in two areas: near the Canavese Line in the Kinzigite Formation paragneisses and in an elongate belt in the Mafic Complex gabbros away from the Canavese Line. 40Ar/39Ar dating indicates that pseudotachylytes from the Kinzigite Formation were formed in the late Eocene, likely during the early stages of the collision between the NW corner of the Adriatic indenter and the European margin. Pseudotachylytes from the Mafic Complex were formed under a transpressive strain regime, as suggested by kinematic indicators, and yield Early Cretaceous (Albian) 40Ar/39Ar ages. Their formation is possibly related to shortening during the inception of the tectonic inversion of the Adriatic passive margin.

Background Gabon is an endemic area for human immunodeficiency virus (HIV) and hepatitis C virus (HCV) and the risk of co-infection is high. Method Between November 2015 and April 2016, we conducted retrospective study on HCV infection among people living with HIV/AIDS (PLHA). A total of 491 PLHA were included in this study and tested for the presence of HCV infection. HIV viral loads were obtained using the Generic HIV viral Load® assay and the CD4+ T cells count was performed using BD FACSCount™ CD4 reagents. HCV screening was performed using the MP Diagnostics HCV ELISA 4.0 kit. HCV genotypes were determined by sequence analysis of NS5B and Core regions. The Mann-Whitney test was used to compare the groups. Chi-2 test and Fisher's Exact Test were used to compare prevalence. Results HCV seroprevalence was 2.9% (14/491), (95% confidence interval (CI):1.4-4.3%). The percentage of HCV viremic patients, defined by the detection of HCV RNA in plasma, was 57% (8/14), representing 1.6% of the total population. HCV seroprevalence and replicative infection were not statistically differ with gender. The percentage of co-infection increased with age. No correlation with CD4+ T cells count and HIV viral load level was registered in this study. Identified HCV strains were predominantly of genotype 4 (87.5%) including 4k, 4e, 4g, 4p, 4f and 4c subtypes. Only one strain belonged to genotype 2 (subtype 2q). Analysis of the NS5B region did not reveal the presence of resistance-associated substitutions for sofosbuvir. Conclusion A systematic screening of hepatitis C is therefore strongly recommended as well as genotyping of HCV strains in order to adapt treatments for the specific case of people living with HIV/AIDS in Central Africa.

We consider differential games with incomplete information. For special games with dynamics independent of the state of the system and linear payoffs, we give a representation formula for the value similar to the value of repeated games with lack of information on both sides. For general games, this representation formula does not hold and we introduce an approximation of the value: we build a sequence of functions converging to the value function.

The prevalence of hepatitis B virus (HBV) surface antigen was significantly higher in urban (12.9%) than in rural (7.6%) populations ( P = 0.003), but no difference was found in the prevalence of hepatitis delta virus (HDV), which was high in both populations. Phylogenetic analysis showed the circulation of HBV-A3 and -E genotypes and the presence of HDV-1, HDV-7, and HDV-8 clades.

To evaluate the efficacy of postexposure prophylaxis with a combination of zidovudine (ZDV), lamivudine (3TC) and indinavir (IDV), after vaginal exposure to HIV.: Experimental intravaginal exposure of female cynomolgus macaques to SIVmac251.ZDV/3TC/IDV treatment was initiated 4 h after exposure and continued for 28 days. Groups of six animals received a placebo or a combination of oral ZDV (4.5 mg/kg), 3TC (2.5 mg/kg) and IDV (20 mg/kg) twice daily or subcutaneous ZDV (4.5 mg/kg) and 3TC (2.5 mg/kg) twice daily, and a higher dose of IDV (60 mg/kg) administered orally twice daily.In the placebo group, all animals were infected. Antiretroviral association protected one of the six animals if all drugs were administered orally and four of the six animals if ZDV and 3TC were administered subcutaneously and IDV was given orally at triple dose. In infected animals, viremia was significantly delayed and lowered in treated animals than in animals given placebo, and high CD4 cell counts were maintained in the treated animals, at least in the medium term. Antiretroviral dosages made in macaques receiving the same treatments showed that protection efficacy could be linked to antiretroviral plasmatic concentration.This study shows, for the first time in macaques, that the postexposure prophylaxis recommended for humans may be effective after vaginal exposure. Improvements in pharmacokinetic parameters significantly increased treatment efficiency.

Hepatitis B virus (HBV) and hepatitis delta virus (HDV) occur worldwide and are prevalent in both urban and remote rural communities. In a remote village in Gabon, central Africa, we observed a high prevalence of HBsAg carriage and HDV infection, particularly in children and adolescents. The prevalence of HBsAg differed significantly by gender and age, females being more likely than males to carry the HBsAg during the first 10 years of life, while the prevalence was higher among males than females aged 11-20 years. We also characterised HBV and HDV strains circulating in the village. The principal HBV strains belonged to genotype HBV-E and subgenotype QS-A3. Complete genome analysis revealed for the first time the presence of the HBV-D genotype in Gabon, in the form of an HBV-D/E recombinant. Molecular analysis of HDV strains and their complete genomic characterisation revealed two distinct groups within the dominant HDV clade 8. Molecular analysis of HBV and HDV strains did not reveal vertical transmission within the families studied but rather horizontal, intrafamilial transmission among children aged 0-10 years. Our findings indicate that HBV is transmitted in early childhood by body fluids rather than by sexual contact. Health education adapted to the different age groups might therefore help to reduce HBV transmission. Young children should be vaccinated to control HBV infection in areas of extremely high prevalence.

Mandrills are the only nonhuman primate (NHP) naturally infected by two types of simian immunodeficiency virus (SIV): SIVmnd-1 and SIVmnd-2. We have already reported that the high SIVmnd-1 replication during primary infection contrasts with only transient changes in CD4 + and CD8 + cell counts. Since early virus-host interactions predict viral control and disease progression in human immunodeficiency virus-infected patients, we investigated the dynamics of SIVmnd-2 primary infection in mandrills to examine the impact on immune effectors in blood and lymph nodes (LNs). To avoid in vitro strain selection, all mandrills in this study received plasma from SIVmnd-2-infected mandrills. SIVmnd-2 plasma viremia peaked at 10 7 to 10 8 RNA copies/ml between days 7 and 10. This peak was followed in all four monkeys by a decline in virus replication, with a set point level of 10 5 to 10 6 RNA copies/ml at day 42 postinfection (p.i.). Viral DNA load in PBMC and LNs also peaked between days 7 and 10 (10 5 to 10 6 DNA copies/10 6 cells) and stabilized at 10 3 to 10 4 DNA copies/10 6 cells during the chronic phase. Anti-SIVmnd-2 antibodies were detected starting from days 28 to 32. A transitory decline of CD3 + CD4 + cells in the LNs occurred in animals with high peak VLs. CD4 + and CD8 + T-cell activation in blood and LNs was noted between days 5 and 17 p.i., surrounding the peak of viral replication. This was most significant in the LNs. Activation markers then returned to preinfection values despite continuous and active viral replication during the chronic infection. The dynamics of SIVmnd-2 infection in mandrills showed a pattern similar to that of SIVmnd-1 infection. This might be a general feature of nonpathogenic SIV natural African NHP models.

An hepatitis B virus (HBV) molecular survey was conducted in five remote villages in the equatorial forest in Gabon, Central Africa. Two hundred seventy out of 311 inhabitants (86.8%) were HBV-infected or had evidence of past HBV infection. Chronic hepatitis corresponding to hepatitis B surface antigen (HBsAg) positivity was suspected in 27 (8.6%) of the HBV-infected subjects. High HBV viral loads were detected mainly in children aged 4-7 years. The pre-S/S domains were sequenced in 13 cases and 12 strains belonged to HBV-A genotype. In one case we found evidence for recombination between genotypes A and E. Phylogenetic analysis revealed that Gabonese HBV strains were distinct from HBV-A subgenotypes (A1 and A2). These new HBV strains from Gabon clustered with previously reported HBV-A3 subgenotype strains from Cameroon and Democratic Republic of Congo. The analysis of the pre-S2 domain allowed us to determine two amino acid substitutions (N/152/S and N/174/T) specific to the Central African HBV-A3 subgenotype strains and one amino acid substitution (P/155/Q) unique to these new Gabonese HBV-A3 subgenotype isolates. Two full genome sequences of two new Gabonese HBV isolates are also presented and confirm the distinctive HBV-Gab-A3 cluster.

Non-invasive faecal sampling in the equatorial forest in Gabon allowed the first identification of the hepatitis B virus (HBV-Ch RC170 ) genome in samples collected from wild chimpanzees ( Pan troglodytes troglodytes ). The HBV-Ch RCl70 sequence clustered with 100% bootstrap support with previous viral sequences obtained from Pan troglodytes subspecies. This is the first evidence of HBV infection in wild apes and confirms that the HBV-like strains thus far characterized in captive apes are directly related to those circulating in the wild.

The mandrill (Mandrillus sphinx) has been shown to be infected with an STLV-1 closely related to HTLV-1. Two distinct STLV-1 subtypes (D and F) infect wild mandrills with high overall prevalence (27.0%) but are different with respect to their phylogenetic relationship and parallel to the mandrills' geographic range. The clustering of these new STLV-1mnd sequences with HTLV-1 subtype D and F suggests first, past simian-to-human transmissions in Central Africa and second, that species barriers are easier to cross over than geographic barriers.

A recent serological survey of wild-born captive monkeys from Gabon, Central Africa, revealed that 1 of 20 Cercopithecus nictitans tested was infected with a new simian T cell lymphotropic virus type 1 (STLV-1). We investigated the molecular relationship between the STLV-1 strain present in this C. nictitans (CN01) and the other available HTLV/STLV strains. Phylogenetic analysis of the env (gp46 and gp21) region showed that the new STLV(nict) clusters with the HTLV-1/STLV-1 group and not with the other nictitans STLVs belonging to the STLV-3 group. Moreover, our new STLV(nict) is closely related to the molecular subtype D, which presently includes five HTLV-1 and three mandrill STLV-1 strains from Gabon and two from Cameroon. These data show that C. nictitans may be the natural carrier of two different molecular types of STLV, one related to STLV-3 and the other possibly one of the simian STLV type 1 counterparts of HTLV-1 subtype D.

Thus far, simian immunodeficiency virus from chimpanzees (SIVcpz) genomes have been characterized as Pan troglodytes troglodytes and show a strong relation with human immunodeficiency virus (HIV)-1 N in their env genes. We fully characterized another SIVcpz from P. t. troglodytes. This chimpanzee (Cam5) was, as was also the host of SIVcpz-cam3, wild born in Cameroon, a region where all three groups of HIV-1 (M, N and O) co-occur. In contrast to other SIVcpz, SIVcpz-cam5 was isolated immediately after the rescue of the animal. Our data demonstrate that SIVcpz-cam5, like SIVcpz-cam3, grows easily on human peripheral blood mononuclear cells (PBMCs) and uses CCR5 as a co-receptor similar to HIV-1 N YBF30. Phylogenetic analysis based on the entire env gene shows that SIVcpz-cam5 falls into the same unique subcluster as HIV-1 N YBF30, SIVcpz-cam3 and SIVcpz-US. A phylogenetic relationship was also found with the vif gene of HIV-1 N. This study provides proof that HIV-1 N related viruses circulate in wild P. t. troglodytes.

African nonhuman primates are natural hosts for simian immunodeficiency viruses (SIVs). To date, SIV infections have been detected in 20 distinct species (24). SIVs were shown to cluster in six major, approximately equidistant lineages (17, 53). Human immunodeficiency virus type 1 (HIV-1) and HIV-2 belong to two of these clusters and emerged most likely following zoonotic transmissions of, respectively, SIVcpz from chimpanzees and SIVsm from sooty mangabeys (14, 16, 21). The remaining clusters are formed by SIVs isolated from African green monkeys (AGMs), Syke's monkeys, l'hoest monkeys, and colobus monkeys (4, 17, 20, 27, 39, 60). The phylogeny of many SIVs resembles that of their host species, suggesting a coevolution (1, 21, 39). In contrast to these, some viruses (SIVrcm, SIVagm.sab, SIVmnd-2, and SIVdrl) cluster in different lineages according to the genomic region analyzed (15, 22, 31, 55). These viruses most likely result from recombination events in monkeys dually infected by SIVs of two distinct lineages (15, 22, 31, 55). Such dual infections suppose the existence of cross-species transmissions of SIV in nature. It has indeed been shown that wild patas monkeys and baboons can acquire SIVagm (7, 32, 61). It remains unclear if these two latter species are infected with species-specific SIVs (54). This was also questioned for mandrills (Mandrillus sphinx). The first SIVmnd strain (GB1), now defined as SIVmnd-1, was isolated 13 years ago from a wild-born mandrill (59). This virus was the only representative of the SIVmnd lineage until two Cercopithecinae viruses, from l'hoest (Cercopithecus l'hoesti l'hoesti) and sun-tailed monkeys (Cercopithecus l'hoesti solatus) were shown to cluster in the same lineage as SIVmnd GB1 (4, 29). As the genome of SIVmnd GB1 had the organization of Cercopithecus SIVs and not that of Papionini SIVs, and because the habitat of mandrills and sun-tailed monkeys overlaps in Gabon, it was suggested that SIVmnd GB1 represents a virus cross-transmitted from sun-tailed monkeys to mandrills, and the lineage was renamed the l'hoest lineage (4, 5). It was subsequently demonstrated that SIVmnd GB1-related viruses are commonly present in wild adult mandrills, indicating that the virus is capable, at least today, of efficiently spreading between animals of this species (55). Mandrills infected with SIVmnd-1 are separated by the Ogooue River from mandrills infected with SIVmnd type 2 (55). This latter virus has a genomic organization distinct from those of SIVmnd GB1 and SIVlhoest and identical to that of SIVs of other Papio monkeys, such as SIVsm and SIVdrl (15, 55). It is unknown why SIV infections are generally nonpathogenic in African nonhuman primates. SIVmnd-2 infections in mandrills have not been associated with signs of AIDS. SIVmnd-1 infection, which apparently results from SIV transmission from C. l'hoesti to mandrills, is also considered to be asymptomatic, although a possible exception has been reported (44). SIVlhoest, which is genetically close to SIVmnd-1 and is also associated with asymptomatic infection in its natural host, appears to induce AIDS in macaques (29). In both HIV-1-infected humans and SIVmac-infected macaques, it was shown that early virus-host interactions are predictive of the outcome of infection. Predictive markers are in particular gag-specific T-helper responses and viral load levels in the post-acute phase of infection (36, 50). In macaques, the steady-state level of plasma viremia 5 to 6 weeks after exposure to the virus is an excellent predictor of the subsequent disease course (28, 35, 42, 63). RNA levels in plasma measured early in HIV-1 infection are also highly predictive of subsequent rates of disease progression (36, 37, 43). However, although this observation is broadly acknowledged and used as the main indication for treating HIV-1-infected patients early in the course of the disease (10), little is still known about the driving mechanism(s) directing this phenomenon. Studies of the early events during nonpathogenic infections in natural host species can help to elucidate such mechanisms. So far, studies during the early phase of infection in African nonhuman primates are limited (58). Studies in SIVagm.sab92018-infected AGMs have revealed for the first time an extensive replication during the acute and post-acute phases (18). Many naturally infected AGMs analyzed during the chronic phase also show continuously high levels of viral RNA in plasma (9, 23). Moreover, virus load in the blood and lymph nodes (LN) of mangabeys during chronic infection is at levels equivalent to that in macaques and humans progressing to AIDS, despite the lack of clinical signs of AIDS (49). The absence of AIDS in these monkeys therefore seems to be paradoxical in the presence of such a high viral load. However, it is not clear whether the high viral replication observed is a general feature of nonpathogenic infections in natural host species (3, 23). Moreover, the precise dynamics of CD4+ and CD8+ cells during primary infection have not been reported so far. Our primary objective for the present study was to investigate the acute and post-acute phases of SIVmnd-1 infection in mandrills. SIVmnd-1 is thought to represent the result of an ancient cross-species transmission in the wild (4, 6), and we investigated whether it would represent an intermediate model between the pathogenic and nonpathogenic models of lentiviral infections. We analyzed viral dynamics and corroborated the virological study with the analyses of CD4+- and CD8+-T-cell changes over time in the blood and LN.

We assess the genetic relationships between 49 HIV-1 group O strains from 24 and 25 patients living in Cameroon and France, respectively. Strains were sequenced in four genomic regions: gag (p24) and three env regions (C2-V3, gp41, and for 22 C2-gp41). In each of the genomic regions analyzed, the genetic diversity among the group O strains was higher than that exhibited by group M. We characterize three major group O phylogenetic clusters (O:A, O:B, and O:C) that comprised the same virus strains in each of the genomic regions analyzed. The majority of strains cluster in O:A, a cluster previously identified by analysis of pol and env sequences. Group O recombinants were also identified. Importantly, the distinction between these three major group O clades was weak compared to the strong clustering apparent in the global group M phylogenetic tree that led to the identification of subtypes. Thus, these clusters of group O viruses should not be considered as equivalent to the group M subtypes. This difference between the pattern of group O and the global group M diversity, both taking into account the pandemic status of the group M subtypes and the comparatively small number of group O-infected individuals (the majority being from Cameroon), indicates that the group O phylogeny primarily represents viral divergence in the Cameroon region, analogous to group M viral diversity present in the Democratic Republic of Congo.

The sensitivity of one ELISA method, six HIV-1/HIV-2 rapid screening tests, and one confirmatory test was evaluated in comparison with a third-generation EIA method (taken as the 'gold standard') and Western blot on well-characterized panels of sera. HIV diversity was represented by 50 HIV-1 group M subtype A to H, nine HIV-1 group O, 12 HIV-2, two HIV1+2 positive and six indeterminate Western blot profiles. Sensitivity during HIV-1 seroconversion was studied on 39 serial samples collected from six patients during early primary infection. Serial samples obtained from two primates during experimental primary SIV infection were used to mimic HIV-2 seroconversion samples. The sensitivity ranged from 100 to 94.6% according to the test. During seroconversion, rapid tests became positive 2-8 days later than the third-generation EIA. This reveals a major limitation of rapid tests, which are being recommended for use in developing countries. The lack of sensitivity seen during early HIV-1 seroconversion and/or limited specificity in some of the evaluated tests present serious limitations to their use in countries with high HIV incidence and variability. It is suggested that, as soon as possible, less sensitive rapid tests for blood bank screening should be abandoned in favor of highly sensitive rapid tests and/or more robust, more sensitive and cheaper ELISAs. These results stress the need for better screening tools and specific local evaluations.

We developed a simple, rapid, inexpensive, and highly sensitive and specific strategy for the detection and lineage differentiation of primate lentiviruses (PIV-ELISA). It is based on the use of two indirect ELISA methods using synthetic peptides mapping the gp41/36 region (detection component) and the V3 region (differentiation component) of four lentivirus lineages, namely SIVcpz/HIV-1 (groups M, O, N, and SIVcpz-gab), SIVmnd, SIVagm, and SIVsm/SIVmac/HIV-2. This strategy was evaluated with panels of sera originating from both humans and nonhuman primates. The human reference panel consisted of 144 HIV Western blot (WB)-positive sera in which the corresponding virus had been genotyped (HIV-1: 72 group M, 28 group O, and 6 group N; HIV-2: 21 subtype A and 10 subtype B; and 7 HIV-1+2) and 105 HIV WB-negative samples. The nonhuman primate reference panel consisted of 24 sera from monkeys infected by viruses belonging to the four lineages included in the PIV-ELISA strategy (5 chimpanzees, 5 macaques, 8 mandrills, and 6 vervets) and 42 samples from seronegative animals. Additional field evaluation panels consisted of 815 human sera from Gabon, Cameroon, and France and 537 samples from 25 nonhuman primate species. All the samples from the two reference panels were correctly detected and discriminated by PIV-ELISA. In the human field evaluation panel, the gp41/36 component correctly identified all the test samples, with 98% specificity. The V3 component discriminated 206 HIV-1 group M, 98 group O, 12 group M+O, and 128 HIV-2 sera. In the primate field evaluation panel, both gp41/36 and V3 detected and discriminated all the WB-positive samples originating from monkeys infected with SIVcpz, SIVagm-ver, SIVmnd-1, SIVmnd-2, SIVdrl, or SIVsun. These results were confirmed by genotyping in every case. Four SIV-infected red-capped mangabeys (confirmed by PCR) were correctly identified by gp41/36, but only two reacted with the V3 peptides in the absence of a specific SIVrcm V3 peptide. Addition of a V3 SIVrcm peptide discriminated all the SIVrcm-positive samples. Fourteen Papio papio samples were positive for SIVsm gp 36 and by WB, but negative by PCR, whereas three Papio cynocephalus samples were positive by gp41/36 but indeterminate by WB and negative by PCR. This combined ELISA system is thus highly sensitive and specific for antibodies directed against HIV and SIV. In addition, the V3-based serotyping results always agreed with genotyping results. This method should prove useful for studies of lentivirus prevalence and diversity in human and nonhuman primates, and may also have the potential to detect previously undescribed SIVs.

Hepatitis B virus (HBV) surface antigen (HBsAg) was found in 9.2% of 1,186 pregnant women from Gabon, of whom 10.1% had the HBe antigen and 89.9% had anti-HBe antibodies. Antibodies to the hepatitis delta virus (HDV) were found in 15.6% of the HBsAg-positive women. The HBV strains were of the A3 and E genotypes. The HDV strains belonged to HDV clades 1 and 8. These results provide clear evidence that HDV clade 8 is indigenous to Africa.

We consider non zero sum two players differential games. We study Nash equilibrium payoffs and publicly correlated equilibrium payoffs. If players use deterministic strategies, it has been proved that the Nash equilibrium payoffs are precisely the reachable and consistent payoffs. Referring to repeated games, we introduce mixed strategies which are probability distributions over pure strategies. We give a characterization of the set of Nash equilibrium payoffs in mixed strategies. Unexpectedly, this set is larger than the closed convex hull of the set of Nash equilibrium payoffs in pure strategies. Finally, we study the set of publicly correlated equilibrium payoffs for differential games and show that it is the same as the set of Nash equilibrium payoffs using mixed strategies.

Background Although a wide variety of non-human primates are susceptible to simian T-cell leukaemia virus type 1 (STLV-1), little is known about the virological or molecular determinants of natural STLV-1 infection. Methods We determined STLV-1 virus tropism in vivo and its relation to the immune response by evaluating cytokine production and T-cell subsets in naturally infected and uninfected mandrills. Results With real-time PCR methods, we found that STLV-1 in mandrills infects both CD4(+) and CD8(+) T cells; however, proviral loads were significantly higher (P = 0.01) in CD4(+) than in CD8(+) cells (mean STLV-1 copies number per 100 cells (+/- SD) was 7.8 +/- 8 in CD4(+) T cells and 3.9 +/- 4.5 in CD8(+) T cells). After culture, STLV-1 provirus was detected in enriched CD4(+) but not in enriched CD8(+) T cells. After 6 months of culture, STLV-1-transformed cell lines expressing CD3(+), CD4(+) and HLADR(+) were established, and STLV-1 proteins and tax/rex mRNA were detected. In STLV-1 infected monkeys, there was a correlation between high proviral load and elevated levels of interleukin (IL)-2, IL-6, IL-10, interferon-gamma and tumour necrosis factor-alpha. The two monkeys with the highest STLV-1 proviral load had activated CD4(+)HLADR(+) and CD8(+)HLADR(+) T-cell subsets and a high percentage of CD25(+) in CD4(+) and CD8(+) T cells. Conclusions Our study provides the first cellular, immunological and virological characterization of natural STLV-1 infection in mandrills and shows that they are an appropriate animal model for further physiopathological studies of the natural history of human T-cell leukaemia viruses.

The mandrill (Mandrillus sphinx) is naturally infected by two types of simian immunodeficiency virus (SIV): SIVmnd types 1 and 2. Both of these viruses cause long-term, non-progressive infections in their natural host despite high plasma viral loads. This study assessed the susceptibility of rhesus macaques to infection by these two types of SIVmnd and compared the virological and basic immunological characteristics of the resulting infections with those observed in natural infection in mandrills. Whilst both SIVmnd types induced similar levels of virus replication during acute infection in both mandrills and macaques, they produced a more pronounced CD4+ T-cell depletion in rhesus macaques that persisted longer during the initial stage of infection. Pro-inflammatory cytokine responses were also induced at higher levels in rhesus macaques early in the infection. During the chronic phase of infection in mandrills, which in this case was followed for up to 2 years after infection, high levels of chronic virus replication did not induce significant changes in CD4+ or CD8+ T-cell counts. In rhesus macaques, the overall chronic virus replication level was lower than in mandrills. At the end of the follow-up period, although the viral loads of SIVmnd-1 and SIVmnd-2 were relatively similar in rhesus macaques, only SIVmnd-1-infected rhesus macaques showed significant CD4+ T-cell depletion, in the context of higher levels of CD4+ and CD8+ T-cell activation, compared with SIVmnd-infected mandrills. The demonstration of the ability of both SIVmnd types to induce persistent infections in rhesus macaques calls for a careful assessment of the potential of these two viruses to emerge as new human pathogens.

Simian immunodeficiency virus (SIV) persistence in wild populations of African nonhuman primates (NHPs) may occur through horizontal and vertical transmission. However, the mechanism(s) and timing of the latter type of transmission have not been investigated to date. Here we present the first study of SIV transmissibility by breast-feeding in an African NHP host. Six mandrill dames were infected with plasma containing 300 50% tissue culture infective doses of SIVmnd-1 on the day after delivery. All female mandrills became infected, as demonstrated by both plasma viral loads (VLs) and anti-SIVmnd-1 seroconversion. Neither fever nor lymphadenopathy was observed. At the peak of SIVmnd-1 viral replication (days 7 to 10 postinoculation), plasma VLs were high (8 × 10 6 to 8 × 10 8 RNA copies/ml) and paralleled the high VLs in milk (4.7 × 10 4 to 5.6 × 10 5 RNA/ml). However, at the end of the breast-feeding period, after 6 months of follow-up, no sign of infection was observed for the offspring. Later on, during a 4-year follow-up examination, two of the offspring showed virological evidence of SIVmnd-1 infection. Both animals seroconverted at least 6 months after the interruption of lactation. In conclusion, despite extensive viral replication in mandrill mothers and high levels of free virus in milk, no SIVmnd-1 transmission was detectable at the time of breast-feeding or during the following months. Since we observed a markedly lower expression of CCR5 on the CD4 + T cells of young mandrills and African green monkeys than on those of adults, we propose that low levels of this viral coreceptor on CD4 + T cells may be involved in the lack of breast-feeding transmission in natural hosts of SIVs.

In order to determine whether geographical or species clustering accounts for the distribution of hepatitis B virus (HBV) in subspecies of chimpanzees in Africa, four complete chimpanzee HBV (ChHBV) genome sequences were obtained from eight hepatitis B surface antigen-positive wild-born chimpanzees from Cameroon, Republic of Congo and Gabon. The serological profiles of these chimpanzees corresponded to the acute or chronic highly replicative phase of HBV infection, as confirmed by high plasma HBV loads. Analysis of the sequence alignment of 256 aa (S region) from the eight HBV-infected chimpanzees allowed us to determine the HBV amino acid patterns specific to each chimpanzee subspecies and to their geographical origin. Phylogenetic analysis of both the S region and the complete genome confirmed this distinctive clustering of eight novel ChHBV strains within Pan troglodytes. The strong phylogenetic associations of ChHBV sequences with both chimpanzee subspecies and their geographical origin were therefore confirmed.

Background Previous epidemiological studies of rural human populations in Gabon reveal a high prevalence of human hepatitis A, B, C and D viruses. In order to investigate the prevalence of the blood-born hepatitis viruses in apes and monkeys living in the same area, we performed an epidemiological survey of HBV, HCV and HDV in wild-born non-human primates. Methods We tested 441 wild-born non-human primates from Gabon and Congo and 132 imported monkeys for the presence of serological markers of HBV, HCV and HDV infections. Results None of Cercopithecidae monkeys were reactive against HBV/HDV and HCV. In contrast, 29.2% of wild-born great apes (154 chimpanzees and 14 gorillas) were positive for HBV serological markers. Nine chimpanzees were in the replicative phase of HBV infection. None of these HBV infected chimpanzees exhibited symptoms or significant changes in serum clinical chemistry related to HBV infection. Conclusions The negativity to HCV-related viruses and the negativity of the Cercopithecidae species tested against HBV/HDV do not allow us to definitively rule out the presence of an animal counterpart of human hepatitis viruses in non-human primates.

We evaluated a new human immunodeficiency virus type 2 (HIV-2) DNA amplification strategy based on peripheral blood mononuclear cell long PCR (XL PCR) followed by nested PCR amplification. The primers used were located in the highly conserved long terminal repeat and in the pol regions of the genome. Five primer pairs corresponding to different regions of the HIV-2 env gene were used in the nested step. Samples from 42 patients were tested, which yielded positive amplification with at least two primer pairs in 40 (95%) samples. A primer pair (EB2/EB5) located on the V3 region succeeded in amplifying proviral DNA in 40 samples.

The three divergent HIV-1 groups M, N and O were very probably introduced into the human population by independent cross-species transmissions of SIVcpz from the chimpanzee subspecies Pan troglodytes troglodytes in central Africa.To characterize HIV-1 group N strains and to elucidate the group's epidemiology and relationship to HIV-1 strains O and M, and SIVcpz.DNA amplification, sequencing and phylogenetic analyses were performed to characterize viruses from three group N-infected individuals (YBF106, YBF115 and YBF116) together with YBF30 and YBF105 previously described.Full-length genome sequence was determined for virus YBF106; gag, pol and env sequences were obtained for YBF116; pol (integrase) and env (gp41) fragments were obtained for YBF115. The gag, pol, 5'-vif and nef sequences were phylogenetically more closely related to HIV-1 M while 3'-vif, vpr, tat, vpu and env clustered with SIVcpz from P. t. troglodytes. Sequence analysis revealed no mutations potentially responsible for drug resistance.The finding that all group N viruses displayed the same recombinant structure and were monophyletic indicates that a single transfer event of SIVcpz to humans can account for the origin of this group. Despite the pathogenic outcome of the known group N infections, the extremely low prevalence of this divergent HIV-1 suggests that this group is not an emerging threat to human health at the present time. However, continuous monitoring of HIV-1 diversity will be important to survey the potential of unusual HIV infections, such as group N, to contribute to the HIV/AIDS pandemic.

Several human and animal Ebola outbreaks have occurred over the past 4 years in Gabon and the Republic of Congo. The human outbreaks consisted of multiple simultaneous epidemics caused by different viral strains, and each epidemic resulted from the handling of a distinct gorilla, chimpanzee, or duiker carcass. These animal populations declined markedly during human Ebola outbreaks, apparently as a result of Ebola infection. Recovered carcasses were infected by a variety of Ebola strains, suggesting that Ebola outbreaks in great apes result from multiple virus introductions from the natural host. Surveillance of animal mortality may help to predict and prevent human Ebola outbreaks.

Hepatitis B and C infections are endemic in human population in central Africa, particularly in Gabon. The aim of this study was to determine the prevalence of hepatitis B virus (HBV) and eventual occurrence of hepatitis C virus (HBC)-related strains in a variety of wild-born non-human primates living in Gabon and Congo. Plasma samples were screened for HBV and HCV markers. A non-invasive method of DNA extraction from faeces followed by specific HBV-DNA amplification was developed to study this infection in wild troops of chimpanzees and gorillas. No HCV infection in non-human primates, wild-born or captive, was detected among 596 samples tested. No HBV infection could be detected in samples tested and obtained from Cercopithecidae. In contrast, 14.7 and 42.2% of wild-born chimpanzees in Gabon and Congo were infected with HBV or had evidence of past HBV infection. At Centre International de Recherches Medicales (CIRMF) Primate Centre, 32.1% of chimpanzees and gorillas were HBV positive or had evidence of past infection. In the cases with past infection, 5.9% wild-born and 8.3% at CIRMF harboured HBV-DNA despite the presence of neutralizing HbsAb. Together with previous findings, we confirm the high HBV prevalence not only in humans but also in chimpanzees and gorillas in Gabon and Congo.

Mandrillus sphinx , a large primate living in Cameroon and Gabon and belonging to the Papionini tribe, was reported to be infected by a simian immunodeficiency virus (SIV) (SIVmndGB1) as early as 1988. Here, we have identified a second, highly divergent SIVmnd (designated SIVmnd-2). Genomic organization differs between the two viral types; SIVmnd-2 has the additional vpx gene, like other SIVs naturally infecting the Papionini tribe (SIVsm and SIVrcm) and in contrast to the other SIVmnd type (here designated SIVmnd-1), which is more closely related to SIVs infecting l'hoest ( Cercopithecus lhoesti lhoesti ) and sun-tailed ( Cercopithecus lhoesti solatus ) monkeys. Importantly, our epidemiological studies indicate a high prevalence of both types of SIVmnd; all 10 sexually mature wild-living monkeys and 3 out of 17 wild-born juveniles tested were infected. The geographic distribution of SIVmnd seems to be distinct for the two types: SIVmnd-1 viruses were exclusively identified in mandrills from central and southern Gabon, whereas SIVmnd-2 viruses were identified in monkeys from northern and western Gabon, as well as in Cameroon. SIVmnd-2 full-length sequence analysis, together with analysis of partial sequences from SIVmnd-1 and SIVmnd-2 from wild-born or wild-living mandrills, shows that the gag and pol regions of SIVmnd-2 are closest to those of SIVrcm, isolated from red-capped mangabeys ( Cercocebus torquatus ), while the env gene is closest to that of SIVmnd-1. pol and env sequence analyses of SIV from a related Papionini species, the drill ( Mandrillus leucophaeus ), shows a closer relationship of SIVdrl to SIVmnd-2 than to SIVmnd-1. Epidemiological surveys of human immunodeficiency virus revealed a case in Cameroon of a human infected by a virus serologically related to SIVmnd, raising the possibility that mandrills represent a viral reservoir for humans similar to sooty mangabeys in Western Africa and chimpanzees in Central Africa.

To determine the prevalence of human immunodeficiency virus type 2 (HIV-2) subtypes circulating in France and to identify possible relationships between these subtypes and pathogenesis, we studied 33 HIV-2-infected patients living in France. HIV-2 DNA was directly amplified from peripheral blood mononuclear cells by nested PCR with specific HIV-2 env primers, and the env gene was sequenced. The serological consequences of antigenic variability were studied by using a panel of peptides and by Western blotting. Phylogenetic analysis classified the 33 HIV-2 strains as subtype A (n = 23) or B (n = 10). There were no significant clinical or epidemiological differences between patients infected with either of these two subtypes. There was some evidence for geographical clustering. Subtype A strains from patients originating from the Cape Verde Islands and Guinea Bissau clustered together. The majority of patients infected with subtype B strains originated from the Ivory Coast or Mali. Strains from patients originating in Mali also clustered in subtype A but distinctly from the Cape Verde or Guinea Bissau strains. The subtype B strains showed greater diversity and included some highly divergent strains relative to those previously characterized. The V3 loop of HIV-2 subtypes A and B was found to be quite conserved in comparison with HIV-1. A strong HIV-2 subtype B serological cross-reactivity was found on HIV-1 env antigen by Western blot mostly in the gp41 transmembrane glycoprotein. This could partly explain the double HIV-1 and HIV-2 reactive profiles found in countries where HIV-2 subtype B is prevalent.

Thus far, simian immunodeficiency virus from chimpanzees (SIVcpz) genomes have been characterized as Pan troglodytes troglodytes and show a strong relation with human immunodeficiency virus (HIV)-1 N in their env genes. We fully characterized another SIVcpz from P. t. troglodytes. This chimpanzee (Cam5) was, as was also the host of SIVcpz-cam3, wild born in Cameroon, a region where all three groups of HIV-1 (M, N and O) co-occur. In contrast to other SIVcpz, SIVcpz-cam5 was isolated immediately after the rescue of the animal. Our data demonstrate that SIVcpz-cam5, like SIVcpz-cam3, grows easily on human peripheral blood mononuclear cells (PBMCs) and uses CCR5 as a co-receptor similar to HIV-1 N YBF30. Phylogenetic analysis based on the entire env gene shows that SIVcpz-cam5 falls into the same unique subcluster as HIV-1 N YBF30, SIVcpz-cam3 and SIVcpz-US. A phylogenetic relationship was also found with the vif gene of HIV-1 N. This study provides proof that HIV-1 N related viruses circulate in wild P. t. troglodytes.

We developed and evaluated two peptide-based immunoassays to confirm and discriminate between group M and group O HIV-1 infection. These assays are based on in vitro competition for antibody binding between M and O peptides. The first EIA is based on competition between group M and group O gp41 immunodominant domains and the second on competition between group O and group M V3 regions of gp120. Two panels of sera were used: the first consisted of 109 sera collected from 27 group O- and 92 group M-infected patients in whom the HIV isolates had been genotyped by sequencing or heteroduplex mobility assay. In this panel, the combination of the two assays correctly discriminated 106 samples (100% group O and 96.7% group M samples). The second panel, used for the field evaluation of the two assays, consisted of 157 samples from HIV-1-infected Cameroonian patients, 33 strains having been genotyped. The combination of the two techniques in a serogrouping algorithm discriminated 147 of these samples, 74 being HIV-1 group O and 73 group M. These results always correlated with genotyping results. The 10 sera that were not successfully classified by these assays were from early seroconverters. Altogether, the two assays clearly differentiated 263 of 276 (94.9%) samples in the two panels. On the basis of the genotyping results, the positive predictive value for group discrimination in the two panels was 100% for both GSEIA assays. Our peptide-blocking group-specific EIAs for differentiation and confirmation of HIV-1 group M and group O infection are complementary tools for epidemiological studies and surveillance of HIV-1 group O strain trafficking.

Le règlement européen relatif aux espèces exotiques envahissantes (EEE) (1143/20141) fournit une liste d’EEE réglementées à l’échelle européenne. A ce jour, sur les 66 EEE de cette liste, seulement deux sont des espèces non indigènes (ENI) marines. L’objectif du présent travail est de proposer une liste d’espèces à réglementation à l’échelle nationale (selon les articles L411-5 dit de Niveau 1 et L411-6 dit de Niveau 2 du Code de l’Environnement2), suivant une nouvelle procédure d’analyse de risques décrite sommairement ci-dessous.