Temporal changes in ewe vaginal microbiota throughout gestation.

Introduction: Numerous factors are known to influence reproductive efficiency in ewes, but few studies have investigated the potential role of vaginal microbiota in sheep reproductive success. The objective of this study was to thoroughly characterize the ewe vaginal microbiota throughout the course of pregnancy. Methods: Vaginal samples were collected from 31 pregnant Hampshire and Hampshire X Suffolk crossbred ewes on a weekly basis from pre-breeding to pregnancy testing and then biweekly until just after lambing. To characterize the vaginal microbial communities, DNA was extracted and 16S rRNA gene Illumina MiSeq amplicon sequencing was performed. Results and Discussion: Alpha diversity metrics indicated an increase in species richness, evenness, and overall diversity throughout gestation. Distinct shifts in the bacterial communities were observed during gestation and were segregated into three periods: early gestation, a transitional period and mid/late gestation. During early gestation, Actinobacillus, Histophilus, and unclassified Leptotrichiaceae were found in greater relative abundance. During the transitional period, a population shift occurred characterized by increasing relative abundance of Streptococcus and Staphylococcus. During mid/late gestation, Staphylococcus, Streptococcus, and Ureaplasma had the greatest relative abundance. These shifts in the microbial population throughout the ewe's gestation are likely related to hormonal changes triggered by the growing conceptus, specifically increasing blood concentration of progesterone. The transitional period shift in vaginal microbial communities potentially aligns with the placental take-over of progesterone production from the corpus luteum at approximately day 50 after conception (gestational week 7). Understanding the observed variability of the vaginal microbiota throughout pregnancy will allow for future comparison of ewes that did not become pregnant or had abnormal pregnancies, which could lead to the discovery of potential bacterial biomarkers for pregnancy outcome; this understanding could also lead to development of probiotics to improve sheep reproductive success.

Erratum: Quantifying and Controlling Entanglement in the Quantum Magnet Cs_{2}CoCl_{4} [Phys. Rev. Lett. 127, 037201 (2021)].

This corrects the article DOI: 10.1103/PhysRevLett.127.037201.

Influence of a sodium-saccharin sweetener on the rumen content and rumen epithelium microbiota in dairy cattle during heat stress.

The effect of a saccharin-based artificial sweetener was tested on animal performance measures and on the microbial communities associated with the rumen content and with the rumen epithelium during heat stress. Ten cannulated Holstein-Friesian milking dairy cattle were supplemented with 2 g of saccharin-based sweetener per day, top-dressed into individual feeders for a 7-day adaptation period followed by a 14-day heat stress period. A control group of ten additional cows subjected to the same environmental conditions but not supplemented with sweetener were included for comparison. 16S rRNA gene amplicon sequencing was performed on rumen content and rumen epithelium samples from all animals, and comparisons of rumen content microbiota and rumen epithelial microbiota were made between supplemented and control populations. Supplementation of the saccharin-based sweetener did not affect the rumen content microbiota, but differences in the rumen epithelial microbiota beta-diversity (PERMANOVA, P = 0.003, R2 = 0.12) and alpha-diversity (Chao species richness, P = 0.06 and Shannon diversity, P = 0.034) were detected between the supplemented and control experimental groups. Despite the changes detected in the microbial community, animal performance metrics including feed intake, milk yield, and short-chain fatty acid (acetic, propionic, and butyric acid) concentrations were not different between experimental groups. Thus, under the conditions applied, supplementation with a saccharin-based sweetener does not appear to affect animal performance under heat stress. Additionally, we detected differences in the rumen epithelial microbiota due to heat stress when comparing initial, prestressed microbial communities to the communities after heat stress. Importantly, the changes occurring in the rumen epithelial microbiota may have implications on barrier integrity, oxygen scavenging, and urease activity. This research adds insight into the impact of saccharin-based sweeteners on the rumen microbiota and the responsivity of the rumen epithelial microbiota to different stimuli, providing novel hypotheses for future research.

Mitigating the effects of heat stress is becoming more and more important with global increases in temperatures. Heat stress negatively affects livestock health and performance. One way to mitigate the effects of heat stress on livestock is to increase feed intake during stress conditions by enhancing palatability of the feed by adding artificial sweeteners. In this study, we investigated whether supplementation of the diet with a saccharin-based sweetener affected dairy cattle performance and the rumen microbial communities during heat stress. We show that supplementation with a saccharin-based artificial sweetener did not affect the performance of the dairy cattle during heat stress. However, the sweetener resulted in changes in the rumen microbial communities, particularly of the microbial communities attached to the rumen wall. These changes in the rumen wall microbial communities could potentially have implications for the host animal, for example in the integrity of the rumen wall barrier function. Future research will be needed to better understand the role of artificial sweeteners in potentially mitigating stress conditions for livestock and to understand their potential effects on microbial communities.

Increased incidence of pathogenic variants in ATM in the context of testing for breast and ovarian cancer predisposition.

Pathogenic Variants (PV) in major cancer predisposition genes are only identified in approximately 10% of patients with Hereditary Breast and Ovarian Cancer (HBOC) syndrome. Next Generation Sequencing (NGS) leads to the characterization of incidental variants in genes other than those known to be associated with HBOC syndrome. The aim of this study was to determine if such incidental PV were specific to a phenotype. The detection rates of HBOC-associated and incidental PV in 1812 patients who underwent genetic testing were compared with rates in control groups FLOSSIES and ExAC. The rates of incidental PV in the PALB2, ATM and CHEK2 genes were significantly increased in the HBOC group compared to controls with, respective odds ratios of 15.2 (95% CI = 5.6-47.6), 9.6 (95% CI = 4.8-19.6) and 2.7 (95% CI = 1.3-5.5). Unsupervised Hierarchical Clustering on Principle Components characterized 3 clusters: by HBOC (P = 0.01); by ExAC and FLOSSIES (P = 0.01 and 0.02 respectively); and by HBOC, ExAC and FLOSSIES (P = 0.01, 0.04 and 0.04 respectively). Interestingly, PALB2 and ATM were grouped in the same statistical cluster defined by the HBOC group, whereas CHEK2 was in a different cluster. We identified co-occurrences of PV in ATM and BRCA genes and confirmed the Manchester Scoring System as a reliable PV predictor tool for BRCA genes but not for ATM or PALB2. This study demonstrates that ATM PV, and to a lesser extent CHEK2 PV, are associated with HBOC syndrome. The co-occurrence of ATM PV with BRCA PV suggests that such ATM variants are not sufficient alone to induce cancer, supporting a multigenism hypothesis.

Evaluating Ruminal and Small Intestinal Morphology and Microbiota Composition of Calves Fed a Macleaya cordata Extract Preparation.

The objective was to determine the impact of feeding MCE on ruminal and intestinal morphology and microbiota composition of calves. A total of 10 male and 10 female crossbred (dairy × beef) calves (6 d of age) were assigned randomly to control (CTL; n = 10) or MCE-supplemented (TRT; n = 10) groups. The MCE was fed in the milk replacer and top-dressed on the calf starter during pre-weaning (6 to 49 d) and post-weaning (50 to 95 d) periods, respectively. Calves were slaughtered at 95 d to collect rumen and intestinal samples to determine volatile fatty acid (VFA) profile, mucosal morphology, and microbiota composition. The effects of MCE were analyzed by accounting for the sex and breed effects. Feeding MCE increased rumen papillae length (p = 0.010) and intestinal villus height: crypt depth (p < 0.030) compared to CTL but did not affect rumen VFA profile. The TRT had a negligible impact on microbial community composition in both the rumen and the jejunum. In conclusion, feeding MCE from birth through weaning can improve ruminal and small intestinal mucosa development of calves despite the negligible microbiota composition changes observed post-weaning.

Quantifying and Controlling Entanglement in the Quantum Magnet Cs_{2}CoCl_{4}.

The lack of methods to experimentally detect and quantify entanglement in quantum matter impedes our ability to identify materials hosting highly entangled phases, such as quantum spin liquids. We thus investigate the feasibility of using inelastic neutron scattering (INS) to implement a model-independent measurement protocol for entanglement based on three entanglement witnesses: one-tangle, two-tangle, and quantum Fisher information (QFI). We perform high-resolution INS measurements on Cs_{2}CoCl_{4}, a close realization of the S=1/2 transverse-field XXZ spin chain, where we can control entanglement using the magnetic field, and compare with density-matrix renormalization group calculations for validation. The three witnesses allow us to infer entanglement properties and make deductions about the quantum state in the material. We find QFI to be a particularly robust experimental probe of entanglement, whereas the one and two-tangles require more careful analysis. Our results lay the foundation for a general entanglement detection protocol for quantum spin systems.

Rumen Epithelial Communities Share a Core Bacterial Microbiota: A Meta-Analysis of 16S rRNA Gene Illumina MiSeq Sequencing Datasets.

In this meta-analysis, 17 rumen epithelial 16S rRNA gene Illumina MiSeq amplicon sequencing data sets were analyzed to identify a core rumen epithelial microbiota and core rumen epithelial OTUs shared between the different studies included. Sequences were quality-filtered and screened for chimeric sequences before performing closed-reference 97% OTU clustering, and de novo 97% OTU clustering. Closed-reference OTU clustering identified the core rumen epithelial OTUs, defined as any OTU present in ≥ 80% of the samples, while the de novo data was randomly subsampled to 10,000 reads per sample to generate phylum- and genus-level distributions and beta diversity metrics. 57 core rumen epithelial OTUs were identified including metabolically important taxa such as Ruminococcus, Butyrivibrio, and other Lachnospiraceae, as well as sulfate-reducing bacteria Desulfobulbus and Desulfovibrio. Two Betaproteobacteria OTUs (Neisseriaceae and Burkholderiaceae) were core rumen epithelial OTUs, in contrast to rumen content where previous literature indicates they are rarely found. Two core OTUs were identified as the methanogenic archaea Methanobrevibacter and Methanomethylophilaceae. These core OTUs are consistently present across the many variables between studies which include different host species, geographic region, diet, age, farm management practice, time of year, hypervariable region sequenced, and more. When considering only cattle samples, the number of core rumen epithelial OTUs expands to 147, highlighting the increased similarity within host species despite geographical location and other variables. De novo OTU clustering revealed highly similar rumen epithelial communities, predominated by Firmicutes, Bacteroidetes, and Proteobacteria at the phylum level which comprised 79.7% of subsampled sequences. The 15 most abundant genera represented an average of 54.5% of sequences in each individual study. These abundant taxa broadly overlap with the core rumen epithelial OTUs, with the exception of Prevotellaceae which were abundant, but not identified within the core OTUs. Our results describe the core and abundant bacteria found in the rumen epithelial environment and will serve as a basis to better understand the composition and function of rumen epithelial communities.

Review: Drinking water for liquid-fed pigs.

Liquid feeding has the potential to provide pigs with sufficient water to remain hydrated and prevent prolonged thirst. However, lack of permanent access to fresh water prevents animals from drinking when they are thirsty. Moreover, individual differences between pigs in a pen may result in uneven distribution of the water provided by the liquid feed, leading to some pigs being unable to meet their water requirements. In this review, we look at the need for and provision of water for liquid-fed pigs in terms of their production performance, behaviour, health and welfare. We highlight factors which may lead to water ingestion above or below requirements. Increases in the need for water may be caused by numerous factors such as morbidity, ambient temperature or competition within the social group, emphasising the necessity of permanent access to water as also prescribed in EU legislation. The drinkers can be the target of redirected behaviour in response to feed restriction or in the absence of rooting materials, thereby generating water losses. The method of water provision and drinker design is critical to ensure easy access to water regardless of the pig's physiological state, and to limit the amount of water used, which does not benefit the pig.

A derivative of ascorbic acid modulates cAMP production.

We reported, in previous experiments, that AA is a global regulator of cAMP pools. In this study, we demonstrate that K873, an analog of AA we synthesized and presenting antiproliferative properties, has also an impact on cAMP production. However, K873 has no antioxidant activity, at the contrary of AA. It definitively demonstrates that action of AA on the cAMP production is not linked to antioxidant activity. These data suggest that AA, and derivatives of this molecule, could be promising drug acting on biological processes that are under the control of cAMP dependent pathway.

Alteration in CD29(high) CD4(+) lymphocyte subset is a common feature of early HIV disease and of active tuberculosis.

Peripheral CD4 T-cell depletion has been observed in human immunodeficiency virus (HIV)-negative patients with pulmonary tuberculosis (TB). To investigate more accurately this alteration, we studied peripheral blood CD45RA(+) and CD29(high) CD4 subsets in 79 TB patients with (HIV(+)TB(+)) or without (HIV(-)TB(+)) HIV infection, 85 HIV-infected patients without TB (HIV(+)TB(-)), and 43 healthy controls, all living in West Africa. The high proportion of CD4(+)CD29(high) T cells observed in controls was dramatically decreased in CDC-A stage HIV(+)TB(-) patients. CD45RA(+) CD4(+) T cells were depleted during the CDC-B stage. Both the percentage and the absolute count of CD29(high)CD4(+) T cells were decreased in HIV(-)TB(+) and HIV(+)TB(+) patients versus controls, but CD45RA(+)CD4(+) T cells were not decreased in TB patients without HIV-infection. Although distinct alterations in the CD4(+) T-cell homeostasis are involved in TB(-) versus HIV-infected subjects, our data suggest that the CD29(+)CD4(+) T-cell depletion observed during the early HIV disease contributes to the risk of active TB, by reducing the pool of T cells able to relocalize to the sites of the M. tuberculosis multiplication.

[Effect of "micronized" C60 fullerene on the microbial growth in vitro]. / Effet du fullerène C60 "micronisé" sur la croissance microbienne in vitro.

The effect of C60 "micronized fullerene was tested by a manual and automatised (Analyzer Bioscreen C(R)) micromethod on the microbial growth of 22 collection strains: E. coli (5), P. aeruginosa (2), S. Typhimurium (6), S. aureus (2), L. monocytogenes (2), E. hirae (1), B. cereus (1), B. subtilis (1), B. pumilus (1) et C. albicans (1). No effect on microbial growth was observed with C60 "micronized fullerene (43.2 microg/ml) on all strains studied: no difference was found with doubling time, slope and growth rate constant. The results of cytotoxicity obtained with animal models or in vitro cultures as human monocyte, leukocyte or macrophage confirm the absence of effect of C60 fullerene at a concentration compatible with microbial or biological models. This study is included in research program headed "Therapeutics perspectives of fullerenes.

Toxicity, mutagenicity, and behavioral effects of beta-CIT, a ligand for dopamine transporter exploration by SPECT.

The cocaine analog beta-CIT is one of the most used compounds for SPET examination of the dopamine transporter in drug abuse and Parkinson's disease. However, the toxicity of this agent has not yet been studied. We report here acute toxicity, mutagenicity, and effect on locomotor activity of beta-CIT. Acute toxicity experiments were performed in mice and rats. The LD50 values were about 20 mg and 5 mg for mice and rats, respectively. There was no sex difference. The mutagenicity was evaluated using the Ames' test. No mutagenic effect was observed for beta-CIT. Effects on locomotor activity were measured in mice using the open-field test. beta-CIT increased locomotion (+65%) when injected at a dose of 0.312 mg/kg; the maximal increase (+205%) was observed at a dose of 1.25 mg/kg; at higher doses, the effect was decreased slightly. These pharmacological findings are in agreement with an inhibitory effect of beta-CIT at the dopamine transporter. We conclude that with no mutagenic effects and LD50 more than 6 orders of magnitude higher than the routinely used doses in PET or SPET, it can be assumed that beta-CIT can be safely used as a radioligand in humans.

[Philippe Maupas : hepatitis B vaccine discoverer]. / Philippe Maupas: inventeur du vaccin contre L'Hépatite B.

Philippe Maupas Day 8th February 1981, an official ceremony with all the Profession, the Tours Faculty of Pharmacy was called Philippe Maupas, Hepatitis B vaccine discoverer - Galien Prize 1981. This communication presents the man, the scientist and the teacher. Born on 30th June 1939 in Toulon (south of France), married and the father of two children, Ph. Maupas was a man of action and an humanist. Full of enthusiasm, always available, passionate about his work, he never hesitated to brave the odds if he felt it would be of use to the community. With a pluridisciplinary training - Veterinary Doctor (1965), Pharmacist (1970), Science Doctor (1970) and Physician Doctor (1976) - he was Professor of Microbiology and Dean of the Faculty of Pharmacy of Tours. His scientific career fully illustrates his thirst for knowledge and his unflagging struggle against infectious diseases. Ph. Maupas approached his research work in a relaxed, imaginative frame of mind. Always passionate about his work and fired by spirit of Louis Pasteur, he was moved by a preoccupation of efficacy and a will of prevention in Public Health. He carried out research into both animal and human infectious diseases as well as anthropozoonosis. Ph. Maupas's most remarkable discoveries concerned the hepatitis B virus: he produced the first vaccine against hepatitis B and applied it to the prevention in man of this disease (1976); he confirmed the aetiological link between the hepatitis B and primary liver cancer.

Impact of short-course therapy on tuberculosis drug resistance in South-West Burkina Faso.

SETTING: The Regional Tuberculosis Centre and the Muraz Centre in Bobo-Dioulasso, Burkina Faso. OBJECTIVES: To observe the trend of primary drug resistance in pulmonary tuberculosis patients 5 years into a short-course treatment programme and to assess the possible implementation of a further programme. DESIGN: Bacteriological study of stains isolated from all newly diagnosed tuberculosis patients (n = 300), all relapse cases (n = 20) and all failure cases (n = 58) from the Houet province, during the period from April 1992 to April 1994. Human immunodeficiency virus (HIV) serostatus was determined for the first 119 patients included in the study. RESULTS: Mycobacterium tuberculosis was the predominant species as shown by 75.1% of the isolates; next was M. africanum, then atypical mycobacteria and finally M. bovis, representing 18.4%, 6.5% and 0.4% of the isolates respectively. Primary resistance (excluding atypical strains) was as follows: isoniazid 7.6%, ethambutol 1.0%, rifampicin 2.5%, and streptomycin 12.4%; 33.6% of the patients tested for HIV were HIV positive. There was no relationship between HIV serostatus and the identity of strains or drug resistance. However, negative acid-fast bacilli smear microscopy with positive culture was significantly more frequent in HIV-positive patients than in HIV-negative patients. CONCLUSION: This study shows a drop in primary resistance compared with previous studies carried out in Bobo-Dioulasso under the same conditions (setting, materials and methods, sampling procedures) in 1982 and 1986. This is consistent with the hypothesis that treatment monitoring and the introduction of short-course therapy in 1989 (2 HERZ/4 HR or 2 HRSZ/4 HR) have contributed to lower rates of primary drug resistance.

Etiologic study of genitourinary infections in women of childbearing age in Bobo-Dioulasso, Burkina Faso, 1992.

BACKGROUND AND OBJECTIVES: Genitourinary infections have a major impact on public health, especially in Africa. Relative distribution of the different pathogens is unknown in Bobo-Dioulasso. GOAL: To describe the etiology of genitourinary infections, to establish the sensitivity of Neisseria gonorrhoeae to antibiotics, and to provide epidemiologic and biologic evidence to optimize the treatment of genitourinary infections. STUDY DESIGN: Clinical and biologic diagnoses were performed on 223 women with genitourinary infections. RESULTS: Etiologies found were trichomoniasis (27.8%), chlamydia (26.9%), bacterial vaginosis (19.7%), candidiasis (16.6), and N. gonorrhoeae infection (10.9%). Human immune deficiency virus antibodies were present in 42% of the patients. Spectinomycin or ceftriaxone should be recommended for the treatment of gonorrhoeae in Bobo-Dioulasso. CONCLUSIONS: The prevalence of Chlamydia trachomatis is higher than that of N. gonorrhoeae in Bobo-Dioulasso. This should be taken into account in clinical management of sexually transmitted diseases in this setting.

PIP: Genitourinary infections have a major impact upon public health, especially in Africa. This paper describes findings from a study conducted to describe the etiology of such infections in Bobo-Dioulasso, to establish the sensitivity of Neisseria gonorrhoeae to antibiotics, and to provide epidemiologic and biologic evidence to optimize the treatment of genitourinary infections. The findings are based upon clinical and biologic diagnoses among 223 women with genitourinary infections. Study found the following etiologies: trichomoniasis in 27.8%, chlamydia in 26.9%, bacterial vaginosis in 19.7%, candidiasis in 16.6%, and Neisseria gonorrhoeae infection in 10.9%. HIV antibodies were present in 42% of patients. The authors recommend spectinomycin or ceftriaxone for the treatment of gonorrhea in Bobo-Dioulasso. Moreover, that the prevalence of Chlamydia trachomatis is higher than that of Neisseria gonorrhoeae should be taken into account when managing STDs in this setting.

Sexually transmitted diseases and human immunodeficiency virus infection among women with genital infections in Burkina Faso.

This study reports the prevalence of sexually transmitted diseases (STDs) among gynaecological outpatients presenting at the Bobo-Dioulasso Hospital (Burkina Faso) with genital infections and examines the factors associated with HIV infection in this population. Of 245 eligible non-pregnant women, 220 consented to participate in the study. Seventy-seven per cent had sexually transmitted infections. The most common were: Trichomonas vaginalis (28%), Chlamydia trachomatis (27%), bacterial vaginosis (20%), Candida albicans (17%), Neisseria gonorrhoeae (11%). The prevalence of HIV infection was 42% (95% c.i. 35.3, 48.3). Logistic regression analyses revealed Neisseria gonorrhoeae to be the only STD significantly associated with infection with HIV (P = 0.04). A sedimentation rate greater than or equal to 100 mm in the first hour was also associated with HIV infection (P < 0.001). Women consulting for genital infections constitute a high risk group for HIV infection and other STDs. Management of these women should focus on the early diagnosis and treatment of STDs.

PIP: During May-October 1992 in the gynecology and obstetrics department at the National Central Hospital Souro Sanou in Bobo-Dioulasso, Burkina Faso, physicians conducted a physical examination of and took vaginal smears from 220 nonpregnant women of reproductive age who consented to take part in this study and who had clinical signs of a genital infection. The researchers wanted to determine the prevalence of sexually transmitted diseases (STDs) and the factors associated with HIV infection. 77% had an STD. The most common STDs were Trichomonas vaginalis (28%) and Chlamydia trachomatis (27%). 42% were HIV positive. HIV-positive women were significantly more likely than HIV-negative women to be infected with Neisseria gonorrhoeae (30.4% vs. 24.2%; p = 0.03). Risk factors associated with HIV infection among women presenting with genital infections included young age (25.5 vs. 27.5 years; p = 0.03), low gravidity (2 vs. 2.7; p = 0.04), a higher sedimentation rate in the first hour (75.3 vs. 54; p 0.001), and a low hemoglobin level (11.7 vs. 12.2 g/dl; p = 0.01). These findings indicate that women with genital infections are a group at high risk of HIV and other STDs and a target population for preventive interventions. Physicians should focus on detection and treatment of STDs when they manage cases with genital infections, and they should give appropriate advice on the prevention of HIV to all women presenting with genital infections.

A study of Toxoplasma and Cytomegalovirus serology in tuberculosis and in HIV-infected patients in Burkina Faso.

Seroreactivity to Toxoplasma gondii (Tg) and to Cytomegalovirus (Cmv) was compared between symptomatic HIV-infected patients (40 with pulmonary tuberculosis and 38 with AIDS) and HIV-seronegative patients (40 tuberculosis patients and 30 healthy patients), in an urban area of Burkina Faso. Prevalence of IgG antibodies to Tg antigens (> 50.0%) did not differ amongst the four groups, but tuberculosis HIV+ patients and AIDS patients showed more higher titers of Tg antibodies more often than healthy patients (p < 0.05 and p < 0.005, respectively). Prevalence of specific IgG to Cmv was higher in tuberculosis HIV-seronegative patients (97.5%) and in AIDS patients (100%) than in healthy patients (82%; p < 0.03 and p < 0.001, respectively). Higher Cmv antibodies titers were found in relation to AIDS but also to tuberculosis. Tuberculosis HIV+ as tuberculosis HIV-patients showed higher Cmv antibodies titers than healthy patients (p < 0.002 and < 0.02 respectively). These data emphasize the need for taking into account the risk of Tg reactivation during the follow-up of HIV infected patients in Burkina Faso and suggest possible relationships between Cmv and tuberculosis reactivations.

Twelve-year follow-up study of hepatitis B immunization of Senegalese infants.

Numerous studies have documented the efficacy and safety of plasma-derived and recombinant hepatitis B vaccines. However, little is known about the long-term protection of hepatitis B vaccine, when anti-HBs declines to low or undetectable levels. This study reports results from a 9-12-year period follow up of infants immunized against hepatitis B in Senegal. At the end of the follow-up period anti-HBs were detected in 81% of children who received a booster dose at school age and in 68% of those who did not. HBsAg was detected in 19% of infants from the control group compared to only 2% of immunized infants, corresponding to a protective efficacy of 88%. The results show that long-term protection against HBsAg carriage of hepatitis B vaccination is very high and that a booster dose at school age does not significantly increase this protection.