Risk factors of exposure to Aedes albopictus bites in mainland France using an immunological biomarker.

In recent decades, the invasive Aedes albopictus vector has spread across Europe and is responsible for numerous outbreaks of autochthonous arboviral disease. The aim of this study was to identify epidemiological and sociological risk factors related to individual levels of exposure to Aedes albopictus bites. A multidisciplinary survey was conducted with volunteer blood donors living in areas either colonised or not by Aedes albopictus in mainland France. Individual levels of exposure were evaluated by measuring the IgG level specific to Aedes albopictus saliva. The most striking risk factors concerned the localisation and characteristics of the dwelling. Individuals living in areas colonised prior to 2009 or recently colonised (between 2010 and 2012) had higher anti-salivary gland extract IgG levels compared with those who were living in areas not yet colonised by Ae. albopictus. The type of dwelling did not seem to impact the level of exposure to Aedes bites. People living in apartments had a higher anti-salivary gland extract IgG level than those living in individual houses but the difference was not statistically significant. Interestingly, the presence of air conditioning or window nets was associated with a noticeable reduction in bite intensity.

Helenalin reduces Staphylococcus aureus infection in vitro and in vivo.

Staphylococcus (S.) aureus is a major udder pathogen causing bovine mastitis. Some pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha), enhance extracellular and intracellular growth of S. aureus, indicating that the inflammatory process favors S. aureus infection. Helenalin is a sesquiterpene lactone with potent anti-inflammatory properties. This study was designed to evaluate the effects of helenalin on S. aureus infection. First, in vitro experiments were conducted. These studies revealed that proliferation of S. aureus in bovine mammary epithelial MAC-T cells treated in the presence or absence of TNF-alpha was markedly reduced in the presence of helenalin. Secondly, in vivo effects of helenalin were investigated. Lactating mice treated in the presence or absence of helenalin were challenged by the intramammary route with S. aureus and the bacteria in the mammary glands were counted 12 h after infection. Significantly less numbers of bacteria were recovered from the infected glands of helenalin-treated mice compared with untreated mice. Moreover, histological examination of mammary tissue from helenalin-treated mice that were challenged with S. aureus indicated that helenalin is able to significantly reduce leukocyte infiltration in the mammary gland following S. aureus inoculation. Our results show that helenalin reduces S. aureus intracellular growth and experimental S. aureus infection. We conclude that helenalin may be of potential interest in the treatment of S. aureus-induced mastitis in the bovine species.

[Experimental infection of goats with Schistosoma bovis and S. curassoni: comparative pathogenic effects]. / Infestation expérimentale de caprins par Schistosoma bovis et S. curassoni: effets pathogènes comparés.

Specific mortality and morbidity have been quantified in goats experimentally infected with Schistosoma bovis or S. curassoni strains from Niger. The study involved nine animals followed during 380 days after infection with, respectively, 1,800 or 2,400 cercariae. S. bovis was significatively more pathogenic than S. curossoni in terms of mortality, weight loss and packed cell volume decrease. In addition, the intensity of clinical symptoms was significatively and positively correlated to the levels of fecal egg excretion. Compared to non-infected controls, a growth differential of, respectively, 1,600 and 880 grams per month should incite to consider S. bovis and S. curassoni as parasites of serious economical impact in sahelian countries.

Schistosoma bovis: vaccine effects of a recombinant homologous glutathione S-transferase in sheep.

The economic importance of the trematode Schistosoma bovis in African livestock has justified the development of a specific vaccine. Administered preventively to sheep, rSb28GST--the only molecule cloned from S. bovis which has demonstrated vaccine potentialities in goats and cattle--reduced the mean worm burden in vaccinated animals and improved their health status compared with that of non-vaccinated controls. As in goats, but not in bovines, the fecundity of the settled worm pairs was not modified. Therefore, rSb28GST can be proposed as a universal tool for the prevention of clinical disorders engendered by the main schistosome species affecting domestic ruminants in the African continent.

Safety and immunogenicity of three doses of a Neisseria meningitidis A + C diphtheria conjugate vaccine in infants from Niger.

BACKGROUND: High rates of endemic disease and recurrent epidemics of serogroup A and C meningococcal meningitis continue to occur in sub-Saharan Africa. A meningococcal A + C polysaccharide diphtheria-toxoid-conjugated vaccine may address this issue. METHODS: In Niger three doses of a bivalent meningococcal A + C diphtheria-toxoid-conjugated vaccine (MenD), containing 1, 4 or 16 microg of each polysaccharide per dose, administered at 6, 10 and 14 weeks of age, were compared with Haemophilus influenzae type b-tetanus toxoid-conjugated (PRP-T) vaccine given with the same schedule or with a meningococcal A + C polysaccharide vaccine (MenPS) given at 10 and 14 weeks of age. One blood sample was taken at the time of enrollment (6 weeks of age) and another was taken 4 weeks after the primary series. RESULTS: All doses of MenD were well-tolerated. After the primary series a higher proportion of infants had detectable serum bactericidal activity against serogroup A for each dose of MenD (from 94% to 100%) than for MenPS (31%) or H. influenzae type b-tetanus toxoid-conjugated vaccine (18.9%); P < or = 0.05. Significant differences were also observed for serogroup C MenD 4 microg or MenD 16 microg (100%) vs. MenPS (69.7%) or Haemophilus influenzae type b-tetanus toxoid-conjugated vaccine (24.3%); P < or = 0.05. When MenPS vaccine was given to 11-month-old children, the immune response measured by both enzyme-linked immunosorbent assay and serum bactericidal assay was greater in those previously immunized with MenD than in those immunized with MenPS vaccine. CONCLUSION: MenD was safe among infants in Niger, and immunization led to significantly greater functional antibody activity than with MenPS. The 4-microg dose of MenD for both the A and C serogroups has been selected for further studies.

Response to conjugate Haemophilus influenzae B vaccine among infants in Niamey, Niger.

Despite near elimination of Haemophilus influenzae b (Hib) meningitis from several industrialized countries following introduction of conjugate Hib vaccines into infant immunization schedules, Hib remains a major cause of meningitis and pneumonia in resource-poor countries. In Niger, Hib causes nearly 200 cases of meningitis per 100,000 children < one year of age, and > 40% of cases are fatal. We evaluated the immunogenicity of Hib polysaccharide-tetanus toxoid conjugate vaccine (PRP-T) administered in the same syringe as diphtheria-tetanus-pertussis (DTP) vaccine among infants in Niger. Infants were randomized into group 1 (PRP-T at six, 10, and 14 weeks), group 2 (PRP-T at 10 and 14 weeks), or a control group (meningococcal A/C polysaccharide vaccine). By 14 weeks of age, all subjects in groups land 2 had > or = 0.15 microg/ml of anti-PRP antibody, and 82% versus 76% had > or = 1.0 microg/ml of antibody (P=not significant). By nine months of age the proportion of infants with > or = 0.15 and > or = 1.0 microg/ml was group I=97% and 76%; group 2=93% and 67%; controls=10% and 2.6%. Four weeks after the first, second, and third doses of PRP-T, infants in group 1 showed geometric mean titers (GMTs) of 0.19, 3.97, and 6.09 microg/ml while infants in group 2 had GMTs of 2.40 and 4.41 microg/ml four weeks after the delayed first and second doses. Both PRP-T groups had significantly higher GMTs at 18 weeks and nine months of age than infants in the control group. The Hib PRP-T vaccine was immunogenic in infants in Niger. The strong response after PRP-T was initiated one month after the first DTP vaccination may reflect carrier priming. Two dose schedules of PRP-T should be given serious consideration, particularly if their reduced cost permits vaccine introduction that would be otherwise unaffordable.

Epidemiological evaluation of a monoclonal ELISA detecting bovine viral diarrhoea pestivirus antigens in field blood samples of persistently infected cattle.

An enzyme-linked immunosorbent assay (ELISA), using monoclonal antibodies for capture and detection, was developed for detecting bovine viral diarrhoea virus (BVDV) antigens in blood samples. The test was evaluated using 761 field samples of known status (viraemic or not). When an appropriate cut-off value was chosen, the sensitivity, specificity, and predictive values of the assay were 100%, higher than the values obtained by classical virus isolation. Correlation with the latter technique exceeded 90%. The ELISA is a good candidate for replacing virus isolation as a reference method for BVDV antigen detection in persistently infected carriers. A method based on the mean of the standard deviation ratio can be used to choose the cut-off value in order to optimise reproducibility.

A monoclonal ELISA for bovine viral diarrhoea pestivirus antigen detection in persistently infected cattle.

Detection of cattle persistently infected with bovine viral diarrhoea virus (BVDV) is crucial to controlling mucosal disease. A sandwich enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies raised against the 48-kDa glycoprotein and the 120/80-kDa protein was developed for detecting antigens in leucocytes of 3 persistently BVDV-infected calves. The test is simple, sensitive and rapid. Moreover the same ELISA was able to recognise Belgian field isolates of BVDV. These results show that the test can be applied in the field.

Use of gel electrophoresis for the study of enzymatic activities of cold-adapted bacteria.

Glutamate dehydrogenase (GDH) and lactate dehydrogenase (LDH) activity of 13 cold-adapted strains, isolated from cold soils and showing GDH and/or LDH activity in spectrophotometric assays, were revealed by the use of electrophoresis on a nondenaturing acrylamide gel (zymogram). Psychrophilic strains were grown at 4 degrees C and 10 degrees C and the psychrotolerant strains at 4 degrees, 20 degrees and 28 degrees C. Incubation with the specific substrate and staining were done at 4, 28 or 37 degrees C. In the most cold-adapted strains, LDH and GDH production was high at 4 degrees C. In psychrotrophic strains, enzyme production and activity were greater at 20 or 28 degrees C than at lower temperatures. LDH remained active up to 37 degrees C while GDH activity was more thermolabile. GDH activity was NAD-dependent in some psychrophilic strains. In other strains, it was dependent on NAD(P) only or on both NAD and NAD(P). Two bands were seen for GDH or LDH activity in some strains. This method, which does not require a dialysis step, can be used to study the influence of temperature on enzyme production and activity, and the co-factor dependence. It detects phenotypic differences between isozymes, providing data for systematics.

Production and characterization of monoclonal antibodies to bovid herpesvirus-4.

Thirty-five hybridoma cell lines secreting monoclonal antibodies (Mabs) against bovid herpesvirus-4 (BHV-4) strain V. Test were produced. These hybrid cells resulted from the fusion of SP2/0 myeloma cells with splenocytes of BALB/c mice previously immunized with purified BHV-4. A modified indirect fluorescent antibody test (IFAT) was applied as a screening procedure and was compared with an indirect enzyme-linked immunosorbent assay. The selected Mabs were tested by the same IFAT against a panel of BHV-4 field isolates and against bovine herpesvirus-1, bovine herpesvirus-2 and alcelaphine herpesvirus-1 (AHV-1). Comparison of BHV-4 field isolates with Mabs confirmed their close antigenic relationships, but slight antigenic differences were observed between different isolates. One of the Mabs also reacted against AHV-1, indicating an antigenic relationship between BHV-4 and AHV-1. None of the Mabs reacting with BHV-4 possessed neutralizing activity against the strain used for immunization.

The oral route as a potential way of transmission of Schistosoma bovis in goats.

The infectivity of Schistosoma bovis cercariae administered orally was evaluated in Sahelian goats. Compared to the percutaneous route, a single massive oral dose resulted in a worm burden and in fecal egg excretion reduced by one-half. Surprisingly, tissue egg counts were increased by more than 4-fold. Fecundity of individual female schistosomes was, therefore, markedly increased. When infective doses were administered weekly for 20 wk, both worm and egg burdens were doubled without modification of the individual worm pair fecundity. Repeated oral infections seem to have induced an acquired tolerance toward parasite antigens. These results confirm the epidemiologic relevance of the oral route in a host species inclined to become infected through drinking water rather than percutaneous exposures.

[Asymptomatic carriage of Pestivirus in ruminants]. / Le portage asymptomatique des Pestivirus chez les ruminants.

Pestiviruses are enveloped single-chain ribonucleic acid viruses with a positive polarity. Pestiviruses include the viruses of classical swine fever (hog cholera), Border disease of sheep, mucosal disease of cattle, and isolates obtained from wild animals, such as red deer (Cervus elaphus). Among ruminants, pestiviruses have developed a remarkable strategy for assuring their persistence. Through epigenetic transmission, they lead to the birth of asymptomatic carrier animals harbouring non-cytopathic variants, which become immunotolerant to the strain of virus present. The presence of a small number of asymptomatic carriers enables the virus to circulate within a herd by horizontal transmission, leading to the birth of a new generation of asymptomatic carriers.

Serological survey for orthopoxvirus infection of wild mammals in areas where a recombinant rabies virus is used to vaccinate foxes.

Several fox vaccination campaigns against rabies have been undertaken in Belgium by using a vaccinia-rabies recombinant virus distributed in baits in the field. However, foxes and other wild animals that may ingest the baits could be infected at the same time by another orthopoxvirus, such as cowpox virus, which circulates in wildlife. Recombination between the two viruses could therefore occur. A serological survey for antibodies to orthopoxvirus, and particularly to cowpox virus, was undertaken in foxes and in several other wild species. Antibodies were detected only in two rodent species, in 16 of 25 bank voles (64 per cent) and in two of 29 woodmice (7 per cent). The risk of virus recombination in wildlife can therefore be considered to be extremely low.

[Reduced morbidity of schistosomiasis: report from an expert workshop on the control of schistosomiasis held at CERMES (15-18 February 2000, Niamey, Niger)]. / Réduire la morbidité des schistosomoses: compte-rendu d'un atelier d'experts sur le contrôle des schistosomoses réunis au CERMES (15-18 février 2000, Niamey, Niger).

Schistosomiasis remains a problem for public health in sub-Saharan Africa. Despite past efforts, cases have not decreased significantly. Schistosoma haematobium and S. mansoni are endemic in all the West African countries. The distribution of both parasites is focal. During a workshop held at CERMES in Niamey, in February 2000, a group of experts recommended that schistosomiasis control be considered as a public health priority in all the endemic West African countries, and National Control Programmes rapidly implemented. The objective of these control programmes would be to reduce schistosomiasis-related morbidity. Case detection should be based on clinical symptoms such as haematuria or bloody diarrhoea, and be carried out at two levels: health care centres and schools, in order to reach patients and school-age children. Health workers should be trained in case detection and community based control of schistosomiasis. The assembled experts advocated the use of praziquantel dosed at 40 mg.kg-1, which therefore must be made available and accessible in outlying areas. Associated measures consist of sanitation, water supply and health education, especially aimed at improving patients' treatment-seeking behaviour. A West African network for schistosomiasis control was created during the workshop. It runs on the Web site of CERMES as network co-ordinator. (http://www.mpl.ird.fr/cermes/).

[Quality control during ultrasonographic evaluation of morbidity due to Schistosoma haematobium in Niger]. / Contrôle de qualité lors de l'évaluation échographique de la morbidité due à Schistosoma haematobium au Niger.

Within the framework of a campaign to control urinary schistosomiasis in Niger, a quality control audit was performed on ultrasonographic assessment of morbidity due to Schistosoma haematobium. The purpose of this audit was to determine variance and reproductibility of epidemiological data provided by two trained independent observers. Three parameters were studied, i.e.,: interobserver variability on matched data, interobserver variance at the community level on the same or different subjects, including some after treatment with praziquantel, and intra-observer variance. A total of 1750 ultrasound examinations were carried out on 1416 inhabitants from 10 hyperendemic villages (70 p. 100 schoolchildren) according to a slightly modified version of the WHO Cairo protocol. Inter-observer variance at the individual level was high for some elementary abnormalities of the bladder. Variance was around 20 p. 100 for the 2 main indicators, i.e. presence of at least one bladder lesion and dilatation of the upper urinary tract. At the community level, inter-observer variance was moderate and the two observers' global assessment of morbidity due to Schistosoma haematobium was the same. Variations of morbidity related to level of endemicity were given perceived in parallel. Similar findings were noted for the intra-observer variability at the individual or community level. Ultrasound examination is supposed to furnish reliable morbidity data for selecting communities at risk and scheduling treatments during schistosomiasis control programs. The results of this study show that the level of inter- and intra-observer variance in ultrasonographic assessment in Niger is compatible with this critical role.

Morphogenesis and release of fowlpox virus.

Release of fowlpox virus (FWPV) as extracellular enveloped virus (EEV) appears to proceed both by the budding of intracellular mature virus (IMV) through the plasma membrane and by the fusion of intracellular enveloped virus (IEV) with the plasma membrane. Based on the frequency of budding events compared to wrapping events observed by electron microscopy, FWPV FP9 strain seems to exit chick embryo fibroblast cells predominantly by budding. In contrast to vaccinia virus (VV), the production of FWPV extracellular virus particles is not affected by N(1)-isonicotinoyl-N(2)-3-methyl-4-chlorobenzoylhydrazine (IMCBH). Comparison of the sequence of the VV F13L gene product with its FWPV orthologue showed a mutation, in the fowlpox protein, at the residue involved in IMCBH resistance in a mutant VV. Glucosamine, monensin or brefeldin A did not have any specific effect on FWPV extracellular virus production. Cytochalasin D, which inhibits the formation of actin filaments, reduces the production of extracellular virus particles by inhibiting the release of cell-associated enveloped virus (CEV) particles from the plasma membrane. Involvement of actin filaments in this mechanism is further supported by the co-localization of actin with viral particles close to the plasma membrane in the absence of cytochalasin D. Actin is also co-localized with virus factories.