There are few studies from developing countries on the epidemiology of Haemophilus influenzae (Hib) infections among infants and children. We set out to determine the prevalence of oropharyngeal Hib colonization among Turkish children younger than two years of age and to identify antimicrobial resistance among the isolates. A cross-sectional study was conducted on 818 healthy children and oropharyngeal secretions were sampled. The carriage rate of Hib was found to be 7·2% and this increased significantly with age. Carriage of Hib among 3- to 6-month-old children (3·5%) was higher than expected and was significantly higher among children who were passive smokers (P=0·04). Logistic regression analysis showed that breastfeeding status was the sole significant factor for colonization (OR 2·2, 95% CI 1·26–3·82). Antimicrobial susceptibility tests on 56 isolates of H. influenzae showed that 51·8% and 21·4% were resistant to trimethoprim–sulphamethoxazole and ampicillin respectively. Other notable resistances were to cefalexin (10·7%) and chloramphenicol (3·6%); no isolates were resistant to ceftriaxone.

Despite a European co-financial programme for control and eradication of brucellosis in Southern Europe, there is evidence that foci of brucellosis still exists in Greece and other Southern European countries. Human brucellosis cases are probably underreported in these countries. A local surveillance system was implemented in a defined region of Northwestern Greece, in order to record and study all human brucellosis cases, using several sources of retrieval. A total of 152 newly diagnosed cases were recorded during a 2-year study period (from 1 April 2002 to 31 March 2004). The age- and sex-adjusted mean annual incidence rate for the population of the study area was 17·3 cases/105 inhabitants. Incomplete application of the control and eradication programme in livestock, and the possible illegal trafficking of animals and their products across the Greek–Albanian border could be responsible for the persistence of foci of brucellosis in the area.

Equine grass sickness (EGS) is a largely fatal, pasture-associated dysautonomia. Although the aetiology of this disease is unknown, there is increasing evidence that Clostridium botulinum type C plays an important role in this condition. The disease is widespread in the United Kingdom, with the highest incidence believed to occur in Scotland. EGS also shows strong seasonal variation (most cases are reported between April and July). Data from histologically confirmed cases of EGS from England and Wales in 1999 and 2000 were collected from UK veterinary diagnostic centres. The data did not represent a complete census of cases, and the proportion of all cases reported to the centres would have varied in space and, independently, in time. We consider the variable reporting of this condition and the appropriateness of the space–time K-function when exploring the spatial-temporal properties of a ‘thinned’ point process. We conclude that such position-dependent under-reporting of EGS does not invalidate the Monte Carlo test for space–time interaction, and find strong evidence for space–time clustering of EGS cases (P<0·001). This may be attributed to contagious or other spatially and temporally localized processes such as local climate and/or pasture management practices.

Herpesviruses can remain dormant in once-infected hosts and, upon reactivation, cause such hosts to become infectious. This phenomenon of latency and reactivation may enable herpesviruses to persist for a long time in small host populations. To quantify the effect of reactivation on persistence, the time to extinction of bovine herpesvirus type 1 (BHV-1) in small cattle populations was calculated. For realistic parameter values the mean time to extinction is already more than 100 years in a population of 10 animals. In a population of 20 animals the time to extinction is approximately 2000 years. The effects of vaccination on persistence were also studied, revealing that continued vaccination of the whole population could result in much faster eradication. For instance, in an isolated herd of 20 animals BHV-1 could be eradicated in 44 years.

The lack of information on the immunity of adults in Brazil against diphtheria prompted us to analyse sera from 234 blood donors aged 18–61 years (30·3% females and 69·7% males). IgG diphtheria antitoxin levels determined by means of an ELISA, validated by toxin neutralization test in Vero cells, showed that 30·7% (95% CI 25·0–37·1) of the population was fully protected ([ges ]1 IU/ml). The highest percentage of subjects fully protected was in the 31–40 years age group. Most of the subjects with uncertain or no protection (<1 IU/ml) were found in the 18–30 years age group (43·8%, OR 2·18, P=0·01). Antitoxin levels were not influenced by the increase in age. Males were more protected than females (80·5%, OR 0·44, P=0·01). The prevalence of 30% of individuals fully protected against diphtheria in blood donors in Rio de Janeiro supports the fact that immunity to diphtheria among healthy Brazilian adults is inadequate. To avoid diphtheria epidemics in the future the immunity among adults should be raised in the coming years.

A prevalence study of verotoxin-producing Escherichia coli O157 (VTEC O157) was performed in 371 randomly selected dairy herds distributed throughout Sweden. Faecal and manure samples were collected and analysed by immunomagnetic separation and culturing. Data were recorded for each herd regarding herd size, age of sampled animals and whether, in addition to cattle, the farm kept other animals. VTEC O157 was isolated from 33 (8·9%) of the 371 investigated herds. The prevalence was higher (23·3%) in Halland county than in the rest of Sweden (P>0·01). Halland was also the county in Sweden that during the study period had the highest incidence of human VTEC O157 cases. VTEC O157 was not detected on any farm in northern Sweden. Identified risk factors, in the multivariate analyses, for herds being VTEC O157 positive were herd size, geographical localization, presence of pigs on the farm and median age of sampled animals.

Between March and July 2003, 671 cases of severe acute respiratory syndrome (SARS) were diagnosed in Taiwan with a total of 84 fatalities. After the epidemic, a serological survey was conducted involving the asymptomatic household contacts. Household contacts of 13 index patients were enrolled in the study. Contact history and clinical symptoms of the household contacts were recorded by standardized questionnaires. Blood samples of patients and household contacts were collected at least 28 days after symptom onset in the index patients or household exposure in the contacts for SARS-associated coronavirus (SARS-CoV) IgG testing. On the basis of this investigation, 29 persons (25 adults and 4 children) were identified as having had unprotected exposure to the index cases before infection-control practices were implemented. Laboratory evaluation of clinical specimens showed no evidence of transmission of SARS-CoV infection to any contacts. This investigation demonstrated that subclinical transmission among household contacts was low in the described setting.

Despite significant control efforts, foot-and-mouth disease (FMD) persists in Turkey, and new strains of serotypes A, O and Asia-1 are periodically reported to enter the country from the east. The status of FMD in Turkey is important regionally because the country forms a natural bridge between Asia where the disease is endemic, and Europe which has disease-free status. This study analysed spatial and temporal patterns of FMD occurrence in Turkey to explore factors associated with the disease's persistence and spread. Annual records of FMD distribution in Turkish provinces throughout 1990–2002, grouped by serotype (O, A and Asia 1), were analysed using geostatistical techniques to explore their spatial and temporal patterns. A meta-population model was used to test how disease status, expressed in terms of presence/absence, extinction, and colonization, and measured at the province level throughout the periods 1990–1996 and 1997–2002, could be predicted using province-level data on: ruminant livestock numbers; meat production-demand discrepancy (as a surrogate measure of animal and animal products marketing, i.e. long-distance contagion through the traffic of mainly live animals to urban centres); and the disease prevalence distribution as recorded for the previous year. A drastic overall reduction in FMD occurrence was observed from the period 1990–1996 to 1997–2002 when the disease was shown to retract into persistence islands. FMD occurrence was associated with host abundance, short distance contagion from adjacent provinces, and meat production-demand discrepancies. With FMD retracting into identified provinces, a shift in predictors of FMD occurrence was observed with a lower contribution of short-distance contagion, and a relatively higher association with meat production-demand discrepancies leading to live animal transport over long distances, and hence presenting opportunities for identifying critical-control points. The pattern of persistence differed according to serotype groups and is discussed in relation to their differential affinity to cattle and small ruminant hosts.

A study was conducted to investigate the persistence of rabbit haemorrhagic disease virus (RHDV) in the environment. Virus was impregnated onto two carrier materials (cotton tape and bovine liver) and exposed to environmental conditions on pasture during autumn in New Zealand. Samples were collected after 1, 10, 44 and 91 days and the viability of the virus was determined by oral inoculation of susceptible 11- to 14-week-old New Zealand White rabbits. Evidence of RHDV infection was based on clinical and pathological signs and/or seroconversion to RHDV. Virus impregnated on cotton tape was viable at 10 days of exposure but not at 44 days, while in bovine liver it was still viable at 91 days. The results of this study suggest that RHDV in animal tissues such as rabbit carcasses can survive for at least 3 months in the field, while virus exposed directly to environmental conditions, such as dried excreted virus, is viable for a period of less than 1 month. Survival of RHDV in the tissues of dead animals could, therefore, provide a persistent reservoir of virus, which could initiate new outbreaks of disease after extended delays.

Pertussis diagnosis among adults is primarily based on serology. In this study, receiver operation characteristics (ROC) analysis was utilized to evaluate three IgA, IgM and IgG ELISAs based on B. pertussis whole-cell sonicate (WC), pertussis toxin (PT), or combined PT-filamentous haemagglutinin (FHA) antigens in a healthy adult population and in adults suffering from symptoms consistent with pertussis. The assays were compared to a PT and FHA reference ELISA. The reference assay showed high correlation with PT- and PT-FHA-based ELISAs but low correlation was found with the WC-based ELISA. However, ROC analysis indicated that all the assays enabled fine differentiation between healthy subjects and subjects suffering from symptoms suggestive of pertussis. The WC IgA ELISA showed the best diagnostic performance [highest specificity (98·8%) with highest sensitivity (53·8%)]. We conclude that despite a low correlation between the various assays they all carry good diagnostic capability for an adult population.

This study investigated whether exposure to inactivated rabbit haemorrhagic disease virus (RHDV) can produce an antigenic response in rabbits and protect them from a subsequent challenge with virulent virus. The aim was to determine if the spreading of baits containing RHDV, which is a common management practice in New Zealand to reduce rabbit numbers, could result in protective immunity in wild rabbits. RHDV was inactivated by ultraviolet (UV) light using an electronic UV crosslinker with a UV dose of 168·48 W-s/cm2 and a UV intensity of 0·0078 W/cm2. Two groups of four rabbits were then inoculated with inactivated virus via oral and intramuscular routes. Rabbits were monitored for 30 days post-inoculation and then challenged orally with virulent virus. No rabbit exposed to inactivated RHDV developed clinical signs of RHD or had antibodies at day 30 post-infection and all animals died within 82 h after challenge with virulent virus. No antibodies were detected at the time of death. These findings suggest that exposure to virus completely inactivated by UV light in the field or on baits will not protect rabbits against challenge with virulent virus.

The occurrence of encephalomyocarditis virus (EMCV) among domestic pigs and wild boar in several European countries is described and discussed. From 1990 to 2001 clinical outbreaks were analysed and serum samples, partly from existing screening programmes, were tested for antibodies against EMCV. Most clinical EMCV outbreaks were reported in Belgium (320), followed by Italy (110), Greece (15) and Cyprus (6). The outbreaks appeared to be clustered in ‘endemic areas’ with an increase in outbreaks during the autumn and winter months. The within-herd seroprevalence measured in clinically affected pig farms varied considerably among farms (2–87%), with age (0–84%) and by country. Data from farms with no clinical disease showed that subclinical infection with EMCV was found both within (seroprevalence 6–62%) and outside (up to 17%) the endemic areas of the clinically affected countries as well as in the non-clinically affected countries Austria and France (3–5·4%). Among wild boar, the seroprevalence varied between 0·6 and 10·8%, and a study in Belgium found a prevalence of virus infection of 3·3%.

Faecal samples of 2660 domestic animals from 116 farms and 956 samples of food were examined for the presence of Shiga toxin-producing Escherichia coli (STEC). STEC was recovered from 126 (15·3%) cattle, 135 (11·3%) pigs, 135 (66·8%) sheep, 31 (73·8%) goats, 4 (1%) chicken, and 15 (1·6%) food samples. Of all STEC isolates, 21·5, 25·8 and 15% produced enterohaemolysin, α-haemolysin, and aerobactin respectively, 1·6% displayed localized adherence (LA) to HEp-2 cells, 27·6% were sorbitol negative, and 30% were resistant to antibiotics. Only 14 (3·1%) of the STEC isolates belonged to human infection-associated serogroups (O26, O55, O111, O128 and O157), designated as enterohaemorrhagic E. coli (EHEC). This study revealed that STEC are prevalent in domestic animals, and to a lesser extent in food of animal origin in Serbia, but the absence of a EHEC phenotypic profile (characteristic serogroup, LA, enterohaemolysin production) in most animal STEC strains may explain the low incidence of human STEC infection in this part of the world.

High titres of pertussis toxin (PT) antibody have been shown to be predictive of recent infection with Bordetella pertussis. The seroprevalence of standardized anti-PT antibody was determined in six Western European countries between 1994 and 1998 and related to historical surveillance and vaccine programme data. Standardized anti-PT titres were calculated for a series of whole-cell and acellular pertussis vaccine trials. For the serological surveys, high-titre sera (>125 units/ml) were distributed throughout all age groups in both high- (>90%) and low-coverage (<90%) countries. High-titre sera were more likely in infants in countries using high-titre-producing vaccines in their primary programme (Italy, 11·5%; Western Germany, 13·3%; France, 4·3%; Eastern Germany, 4·0%) compared to other countries (The Netherlands, 0·5%; Finland, 0%). Recent infection was significantly more likely in adolescents (10–19 years old) and adults in high-coverage countries (Finland, The Netherlands, France, East Germany), whereas infection was more likely in children (3–9 years old) than adolescents in low-coverage (<90%; Italy, West Germany, United Kingdom) countries. The impact and role of programmatic changes introduced after these surveys aimed at protecting infants from severe disease by accelerating the primary schedule or vaccinating older children and adolescents with booster doses can be evaluated with this approach.

A summer camp was followed by an outbreak of illness involving around 90 children. Investigations included individual questionnaires, inspection of the camp facilities, and laboratory analysis of water and clinical samples. Contamination of drinking and swimming water was demonstrated. An enterovirus was detected by polymerase chain reaction (PCR) and/or culture in 4/4 cerebrospinal fluid samples, 9/15 (60%) stool samples from symptomatic children and 2/9 (22%) stool samples from asymptomatic children. The virus was identified as an echovirus 3 by sequencing and phylogenetic analysis of a short 5′ non-coding region (NCR) PCR product. Viruses from the outbreak clustered closely and an echovirus 3 from a temporally associated non-outbreak case could be readily distinguished. Despite the lack of a standardized approach, direct molecular detection and identification of enteroviruses is an efficient epidemiological tool. Here the 5′-NCR was successfully used for both detection and ‘serotyping’, and the close genetic relatedness of isolates was proven.

The prevalence of specific HSV-2 antibodies was studied in Tanzanian and Norwegian sexually transmitted disease (STD) patients (1095) and non-STD patients (488). Correlates to demographic and behavioural factors were evaluated. Seropositivity was determined by the non-commercial peptide-55 enzyme-linked immunoassay. The prevalence of HSV-2 antibodies was 70% in Tanzanian and 17% in Norwegian STD patients, 35% in Tanzanian blood donors and pregnant women, and 4, 7 and 14% in Norwegian medical students, blood donors and pregnant women respectively. A higher HSV-2 prevalence was associated with female sex, increasing age, previous STDs, history of genital HSV infection, coitarchal age (age at first intercourse) <15 years and HIV seropositivity. Compared to previous data, the prevalence of HSV-2 antibodies in Tanzanian STD patients has increased remarkably. In Norwegian STD patients our results are consistent with, or lower than, the prevalence previously reported in Western Europe. Demographic rather than behavioural factors were associated with higher prevalence of HSV-2 antibodies in STD patients.

Approximately 42% (187/444) of swine enterococci collected between the years 1999 and 2000 exhibited high-level resistance to gentamicin (MIC [ges ]500 μg/ml), kanamycin (MIC [ges ]500 μg/ml), or streptomycin (MIC [ges ]1000 μg/ml). Eight aminoglycoside resistance genes were detected using PCR, most frequently ant(6)-Ia and aac(6′)-Ii from Enterococcus faecium. Twenty-four per cent (45/187) of total high-level aminoglycoside-resistant isolates and 26% (4/15) of isolates resistant to high levels of all three antimicrobials were negative for all genes tested. These data suggest that enterococci isolated from swine contain diverse and possibly unidentified aminoglycoside resistance genes.

A reverse transcription–polymerase chain reaction (RT–PCR), that uses primers specifically designed to amplify a portion of the N gene of vampire bat strains of rabies that circulate in Mexico, but also recognizing most of the rabies variants circulating in endemic areas, was established. This standardized PCR assay was able to detect viral RNA in tenfold serial dilutions up to a 107 dilution using stock virus at an original titre of 107·5 LD50. The assay was highly specific for rabies virus. Forty different rabies isolates recovered from different species and geographical regions in the country were diagnosed as positive and negative by the fluorescent antibody test (FAT). These same samples were re-examined by both PCR and the mouse inoculation test (MIT). Compared with MIT the PCR exhibited an epidemiological sensitivity of 86% and a specificity of 91% while its positive predictive value was 96%.

Although veterinary authorities aim to limit persistence of classical swine fever (CSF) in wild boar (Sus scrofa), to avoid potential transmission to pigs, factors influencing CSF transmission and persistence are not clearly understood. Here we analyse incidence and persistence in a CSF epidemic that occurred in the French Vosges Forest. Higher incidence was found in large forests compared to smaller isolated ones, being highest near the starting point of the epidemic, but poorly related to the local density. We hypothesize that the spatial and social structure of wild boar populations may be responsible for this variability of incidence over space. Persistence was highest near the starting point of the epidemic and where initial density was highest. We hypothesize that persistence was favoured by the abundance of young wild boar, itself encouraged by CSF. Our results allow us to propose management measures aimed at limiting CSF persistence.

To investigate the possible dependence on age of the rate of decline of IgG antibodies to pertussis toxin (IgG-PT) after natural infection with Bordetella pertussis we measured IgG-PT in follow-up sera of 121 patients (age 0–94 years) obtained after 123 episodes of B. pertussis infection. For analysis we applied a dynamic model for the inactivation of B. pertussis by the immune system. There were no significant differences in rise, peak and decline of IgG-PT between different age groups, although there was a tendency for a more rapid increase, a higher peak and a faster decline with increasing age. The IgG-PT cut-off of 100 U/ml for serodiagnosis of pertussis appeared valid in all age groups. A decline of IgG-PT to <10 U/ml was associated with increased risk of re-infection with B. pertussis.