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Background Leptospirosis diagnoses have increased in Sarawak, Malaysia in recent years. Methods To better understand the burden of disease and associated risk factors, we evaluated 147 patients presenting with clinical leptospirosis to local hospitals in Sarawak, Malaysia for the presence of Leptospira and associated antibodies. Sera and urine specimens collected during the acute illness phase were assessed via a commercially available rapid diagnostic test (Leptorapide, Linnodee Ltd., Antrim, Northern Ireland), an ELISA IgM assay (Leptospira IgM ELISA, PanBio, Queensland, Australia) and a pan-Leptospira real-time PCR (qPCR) assay to estimate disease prevalence and diagnostic accuracy of each method. Microagglutination testing was performed on a subset of samples. Results Overall, 45 out of 147 patients (30.6%) showed evidence of leptospires through qPCR in either one or both sera (20 patients) or urine (33 patients), and an additional ten (6.8%) were considered positive through serological testing, for an overall prevalence of 37.4% within the study population. However, each diagnostic method individually yielded disparate prevalence estimates: rapid test 42.2% for sera and 30.5% for urine, ELISA 15.0% for sera, qPCR 13.8% for sera and 23.4% for urine. Molecular characterization of a subset of positive samples by conventional PCR identified the bacterial species as Leptospira interrogans in 4 specimens. A multivariate risk factor analysis for the outcome of leptospirosis identified having completed primary school (OR = 2.5; 95 CI% 1.0–6.4) and weekly clothes-washing in local rivers (OR = 10.6; 95 CI% 1.4–214.8) with increased likelihood of leptospirosis when compared with those who had not. Conclusion Overall, the data suggest a relatively high prevalence of leptospirosis in the study population. The low sensitivities of the rapid diagnostic test and ELISA assay against qPCR highlight a need for better screening tools.

Leptospirosis is the most common bacterial zoonosis globally. The pathogen, Leptospira spp., is primarily associated with rodent reservoirs. However, a wide range of other species has been implicated as reservoirs or dead-end hosts. We conducted a survey for Leptospira spp. in bats and rodents from Papua New Guinea. Kidney samples were collected from 97 pteropodid bats (five species), 37 insectivorous bats from four different families (six species) and 188 rodents (two species). Leptospires were detected in a high proportion of pteropodid bats, including Nyctimene cf. albiventer (35%), Macroglossus minimus (34%) and Rousettus amplexicaudatus (36%). Partial sequencing of the secY gene from rodent and bat leptospires showed host species clustering, with Leptospira interrogans and L. weilii detected in rodents and L. kirschneri and a potential novel species of Leptospira detected in bats. Further research is needed in Papua New Guinea and other locales in the Pacific region to gain a better understanding of the circulation dynamics of leptospires in reservoir species and the risks to public and veterinary health.

Glanders is a contagious zoonotic disease caused by Burkholderia mallei. Following the detection of glanders positive horses using the OIE complement fixation test, the tissues of two horses were analysed by PCR. While PCR systems targeting the Burkholderia pseudomallei complex gave positive signals, the species-specific PCR systems targeting B. mallei (fliP-IS407A) and B. pseudomallei (orf11)-the OIE recommended targets-resulted in negative signals. However, the presence of B. mallei in these tissues was confirmed with a recently described B. mallei-specific real-time PCR system and genotyping with MLST- and SNP-based methods, performed on the most positive tissue, identified a genotype closely related to B. mallei strains recently isolated in the Middle East. This study leads to recommendations regarding the use of PCR systems for the molecular diagnosis of glanders, especially in regions where the circulating B. mallei strains have not yet been fully genetically characterized.

Antibiotic-resistant Campylobacter constitutes a serious threat to public health. The clonal expansion of resistant strains and/or the horizontal spread of resistance genes to other strains and species can hinder the clinical effectiveness of antibiotics to treat severe campylobacteriosis. Still, gaps exist in our understanding of the risks of acquisition and spread of antibiotic resistance in Campylobacter. While the in vitro transfer of antimicrobial resistance genes between Campylobacter species via natural transformation has been extensively demonstrated, experimental studies have favored the use of naked DNA to obtain transformants. In this study, we used experimental designs closer to real-world conditions to evaluate the possible transfer of antimicrobial resistance genes between Campylobacter strains of the same or different species (Campylobacter coli or Campylobacter jejuni) and originating from different animal hosts (swine or turkeys). This was evaluated in vitro through co-culture experiments and in vivo with dual-strain inoculation of turkeys, followed by whole genome sequencing of parental and newly emerged strains. In vitro, we observed four independent horizontal gene transfer events leading to the acquisition of resistance to beta-lactams (blaOXA), aminoglycosides [aph(2′′)-If and rpsL] and tetracycline [tet(O)]. Observed events involved the displacement of resistance-associated genes by a mutated version, or the acquisition of genomic islands harboring a resistance determinant by homologous recombination; we did not detect the transfer of resistance-carrying plasmids even though they were present in some strains. In vivo, we recovered a newly emerged strain with dual-resistance pattern and identified the replacement of an existing non-functional tet(O) by a functional tet(O) in the recipient strain. Whole genome comparisons allowed characterization of the events involved in the horizontal spread of resistance genes between Campylobacter following in vitro co-culture and in vivo dual inoculation. Our study also highlights the potential for antimicrobial resistance transfer across Campylobacter species originating from turkeys and swine, which may have implications for farms hosting both species in close proximity.

The gut microbiota is an immense reservoir of antimicrobial resistance genes (ARGs), the so-called “resistome.” In Australia, where antibiotic use is high and resistance rates in some common pathogens are increasing, very little is known about the human resistome. To assess the presence and diversity of ARGs in the gut of Australians from south-eastern Victoria, we investigated fecal samples from clinically healthy infants and pregnant women using non-targeted (shotgun metagenomics sequencing or SMS) and targeted sequencing (two Ion AmpliseqTM panels). All methods detected ARGs in all samples, with the detection overall of 64 unique genes conferring resistance to 12 classes of antibiotics. Predominant ARGs belonged to three classes of antibiotics that are the most frequently prescribed in Australia: tetracycline, β-lactams and MLSB (macrolide, lincosamide, streptogramin B). The three bacterial Orders commonly identified as carrying ARGs were Clostridiales, Bacteroidales, and Enterobacteriales. Our preliminary results indicate that ARGs are ubiquitously present and diverse among the gut microbiota of clinically healthy humans from south-eastern Victoria, Australia. The observed resistance pattern partly overlaps with antimicrobial usage in human medicine in Australia, but ARGs to tetracycline are more common than could be expected. Our current sample is small and limited to south-eastern Victoria, and more data on healthy individuals will be needed to better depict resistance patterns at the population level, which could guide population and/or environmental monitoring and surveillance of antibiotic resistance on various spatio-temporal scales in Australia. For future studies, we recommend using the Ion AmpliseqTM Antimicrobial Resistance Research panel, which is sensitive and user-friendly, or combining several methods to increase the detected diversity.

Leptospirosis, an environmental infectious disease of bacterial origin, is the infectious disease with the highest associated mortality in Seychelles. In small island territories, the occurrence of the disease is spatially heterogeneous and a better understanding of the environmental factors that contribute to the presence of the bacteria would help implement targeted control. The present study aimed at identifying the main environmental parameters correlated with animal reservoirs distribution and Leptospira infection in order to delineate habitats with highest prevalence. We used a previously published dataset produced from a large collection of rodents trapped during the dry and wet seasons in most habitats of Mah&eacute, the main island of Seychelles. A land use/land cover analysis was realized in order to describe the various environments using SPOT-5 images by remote sensing (object-based image analysis). At each sampling site, landscape indices were calculated and combined with other geographical parameters together with rainfall records to be used in a multivariate statistical analysis. Several environmental factors were found to be associated with the carriage of leptospires in Rattus rattus and Rattus norvegicus, namely low elevations, fragmented landscapes, the proximity of urbanized areas, an increased distance from forests and, above all, increased precipitation in the three months preceding trapping. The analysis indicated that Leptospira renal carriage could be predicted using the species identification and a description of landscape fragmentation and rainfall, with infection prevalence being positively correlated with these two environmental variables. This model may help decision makers in implementing policies affecting urban landscapes and/or in balancing conservation efforts when designing pest control strategies that should also aim at reducing human contact with Leptospira-laden rats while limiting their impact on the autochthonous fauna.

Objective: Hepatitis E virus (HEV) has been scarcely investigated in the Indian Ocean. Following a nationwide serosurvey among blood donors, we conducted a population-based serosurvey to assess the magnitude of HEV exposure on Reunion Island. Methods: Four hundred and sixty-six archived frozen human sera from the 2009 CoPanFlu-RUN cohort were analysed using the Wantai HEV IgG enzyme immunoassay. HEV seropositivity was defined as an IgG titre ≥5 UI/ml. Raw and weighted seroprevalences were assessed to account for the discrepancy between the CoPanFlu-RUN subset and the general community. Prevalence proportion ratios (PPR) were measured using log-binomial models. Results: The raw and the weighted seroprevalences of HEV were 9.01% (95% CI 6.41–11.61) and 6.73% (95% CI 4.47–8.98), respectively. The presence of HEV IgG antibodies was associated with increasing age (P

The purpose of the study was to weigh the community burden of chikungunya determinants on Reunion island. Risk factors were investigated within a subset of 2101 adult persons from a population-based cross-sectional serosurvey, using Poisson regression models for dichotomous outcomes. Design-based risk ratios and population attributable fractions (PAF) were generated distinguishing individual and contextual (i.e. that affect individuals collectively) determinants. The disease burden attributable to contextual determinants was twice that of individual determinants (overall PAF value 89.5% vs. 44.1%). In a model regrouping both categories of determinants, the independent risk factors were by decreasing PAF values: an interaction term between the reporting of a chikungunya history in the neighbourhood and individual house (PAF 45.9%), a maximal temperature of the month preceding the infection higher than 28.5 °C (PAF 25.7%), a socio-economically disadvantaged neighbourhood (PAF 19.0%), altitude of dwelling (PAF 13.1%), cumulated rainfalls of the month preceding the infection higher than 65 mm (PAF 12.6%), occupational inactivity (PAF 11.6%), poor knowledge on chikungunya transmission (PAF 7.3%) and obesity/overweight (PAF 5.2%). Taken together, these covariates and their underlying causative factors uncovered 80.8% of chikungunya at population level. Our findings lend support to a major role of contextual risk factors in chikungunya virus outbreaks.

Background Tuberculosis (TB) is a serious health problem in Papua New Guinea (PNG) with an estimated 30000 new cases and 3800 deaths each year. In the Balimo region of the Western Province, diagnosis relies on clinical manifestations and on the microscopic detection of acid-fast bacilli (AFB) in sputum smears, a technique with limited sensitivity. Methods A molecular diagnosis assay targeting DNA extracted from archived sputum smear slides collected from the Balimo region (2012–2014) was conducted, without the need for a viable culture. The presence of Mycobacterium sp on 1162 slides prepared from 345 sputum samples was assessed using a real-time PCR (qPCR) approach. Results The qPCR technique identified the presence of mycobacteria in 35.4% of the smear slides and 59.7% of the tested sputum samples. Poor agreement was observed between the two diagnosis methods (smear AFB microscopy versus qPCR), with 100 AFB-positive sputum samples compared to 206 qPCR-positive sputum samples overall. Treatment was initiated in 90.2% of the smear-positive cases. Unnecessary treatment of ‘false-positive' TB cases (AFB-negative/qPCR-negative) was very low (8.6%) and was even lower when the nine patients diagnosed with extrapulmonary TB were excluded from the analysis. However, the prevalence of false-negatives (AFB-negative/qPCR-positive) was high (28.5%). Conclusions Undetected smear-negative TB is occurring in the Balimo region of PNG, as well as some unnecessary empirical treatment. Molecular methods of diagnosis could greatly reduce the frequency of inappropriate clinical assessment, as well as providing point-of-care diagnosis. This may provide substantial patient and programmatic benefits, including lowering the economic burden on patients from rural areas seeking medical diagnosis in Balimo.

Objective Murine typhus has been increasingly reported on Reunion island, Indian ocean, following documentation of eight autochthonous infections in 2012–2013. We conducted a serosurvey to assess the magnitude of the seroprevalence of rickettsioses in the population. Two hundred and forty-one stored frozen sera taken from the 2009 Copanflu-RUN cohort were analysed using an immunofluorescence assay allowing to distinguish typhus group (TGR) and spotted fever group Rickesttsiae (SFGR). Seropositivity was defined for a dilution titre of Rickettsia IgG antibodies ≥ 1:64. Seroprevalence was weighted to account for the discrepancy between the Copanflu-RUN subset and the general population, as to infer prevalence at community level. Prevalence proportion ratios (PPR) were measured using log-binomial models. Results The weighted seroprevalences of typhus group rickettsioses and spotted fever group rickettsioses were of 12.71% (95% CI 8.84–16.58%) and 17.68% (95% CI 13.25–22.11%), respectively. Pooled together, data suggested that a fifth of the population had been exposed at least to one Rickettsia group. Youths (

After the documentation of sporadic cases of Q fever endocarditis, we conducted a serosurvey to assess Coxiella burnetii exposure on Reunion Island. Two hundred forty-one stored frozen human sera were analyzed using an immunofluorescence assay. The weighted seroprevalence of Q fever was of 6.81% (95% confidence interval, 4.02%–9.59%). Despite the absence of infection in youths

Papua New Guinea (PNG) has a high burden of tuberculosis (TB), including drug-resistant TB (DR-TB). DR-TB has been identified in patients in Western Province, although there has been limited study outside the provincial capital of Daru. This study focuses on the Balimo region of Western Province, aiming to identify the proportion of DR-TB, and characterise Mycobacterium tuberculosis (MTB) drug resistance-associated gene mutations. Sputum samples were investigated for MTB infection using published molecular methods. DNA from MTB-positive samples was amplified and sequenced, targeting the rpoB and katG genes to identify mutations associated with rifampicin and isoniazid resistance respectively. A total of 240 sputum samples were collected at Balimo District Hospital (BDH). Of these, 86 were classified as positive based on the results of the molecular assays. For samples where rpoB sequencing was successful, 10.0% (5/50, 95% CI 4.4⁻21.4%) were considered rifampicin-resistant through detection of drug resistance-associated mutations. We have identified high rates of presumptive DR-TB in the Balimo region of Western Province, PNG. These results emphasise the importance of further surveillance, and strengthening of diagnostic and treatment services at BDH and throughout Western Province, to facilitate detection and treatment of DR-TB, and limit transmission in this setting.

Tuberculosis remains the world’s leading cause of death from an infectious agent, and is a serious health problem in Papua New Guinea (PNG) with an estimated 36,000 new cases each year. This study describes the genetic diversity of Mycobacterium tuberculosis among tuberculosis patients in the Balimo/Bamu region in the Middle Fly District of Western Province in PNG, and investigates rifampicin resistance-associated mutations. Archived Ziehl-Neelsen-stained sputum smears were used to conduct microbead-based spoligotyping and assess genotypic resistance. Among the 162 samples included, 80 (49.4%) generated spoligotyping patterns (n = 23), belonging predominantly to the L2 Lineage (44%) and the L4 Lineage (30%). This is consistent with what has been found in other PNG regions geographically distant from Middle Fly District of Western Province, but is different from neighbouring South-East Asian countries. Rifampicin resistance was identified in 7.8% of the successfully sequenced samples, with all resistant samples belonging to the L2/Beijing Lineage. A high prevalence of mixed L2/L4 profiles was suggestive of polyclonal infection in the region, although this would need to be confirmed. The method described here could be a game-changer in resource-limited countries where large numbers of archived smear slides could be used for retrospective (and prospective) studies of M. tuberculosis genetic epidemiology.

Understanding the processes driving parasite assemblages is particularly important in the context of zoonotic infectious diseases. Leptospirosis is a widespread zoonotic bacterial infection caused by pathogenic species of the genus Leptospira. Despite a wide range of animal hosts, information is still lacking on the factors shaping Leptospira diversity in wild animal communities, especially in regions, such as tropical insular ecosystems, with high host species richness and complex biogeographical patterns. Using a large dataset (34 mammal species) and a multilocus approach at a regional scale, we analyzed the role of both host species diversity and geography in Leptospira genetic diversity in terrestrial small mammals (rodents, tenrecs, and shrews) and bats from 10 different islands/countries in the western Indian Ocean (WIO) and neighboring Africa. At least four Leptospira spp. (L. interrogans, L. borgpetersenii, L. kirschneri, and L. mayottensis) and several yet-unidentified genetic clades contributed to a remarkable regional Leptospira diversity, which was generally related to the local occurrence of the host species rather than the geography. In addition, the genetic structure patterns varied between Leptospira spp., suggesting different evolutionary histories in the region, which might reflect both in situ diversification of native mammals (for L. borgpetersenii) and the more recent introduction of non-native host species (for L. interrogans). Our data also suggested that host shifts occurred between bats and rodents, but further investigations are needed to determine how host ecology may influence these events.

Background Bidirectional signalling between the brain and the gastrointestinal tract is regulated at neural, hormonal, and immunological levels. Recent studies have shown that helminth infections can alter the normal gut microbiota. Studies have also shown that the gut microbiota is instrumental in the normal development, maturation and function of the brain. The pathophysiological pathways by which helminth infections contribute to altered cognitive function remain poorly understood. Discussion We put forward the hypothesis that gastrointestinal infections with parasitic worms, such as helminths, induce an imbalance of the gut-brain axis, which, in turn, can detrimentally manifest in brain development. Factors supporting this hypothesis are: 1) research focusing on intelligence and school performance in school-aged children has shown helminth infections to be associated with cognitive impairment, 2) disturbances in gut microbiota have been shown to be associated with important cognitive developmental effects, and 3) helminth infections have been shown to alter the gut microbiota structure. Evidence on the complex interactions between extrinsic (parasite) and intrinsic (host-derived) factors has been synthesised and discussed. Summary While evidence in favour of the helminth-gut microbiota-central nervous system hypothesis is circumstantial, it would be unwise to rule it out as a possible mechanism by which gastrointestinal helminth infections induce childhood cognitive morbidity. Further empirical studies are necessary to test an indirect effect of helminth infections on the modulation of mood and behaviour through its effects on the gut microbiota.

SUMMARYLeptospirosis is a zoonotic bacterial disease of global importance. A large spectrum of asymptomatic animal hosts can carry the infection and contribute to the burden of human disease. Environmental sources of human contamination also point to the importance of a hydrotelluric reservoir. Leptospirosis can be caused by as many as 15 different pathogenic or intermediateLeptospiraspecies. However, classification of these bacteria remains complicated through the use of both serological and genetic classification systems that show poor correlation. With the advent of molecular techniques, DNA-based barcoding offers a conceptual framework that can be used for leptospirosis surveillance as well as source tracking. In this review, we summarize some of the current techniques, highlight significant successes and weaknesses and point to the future opportunities and challenges to successfully establish a widely applicable barcoding scheme forLeptospira.

Background Leptospirosis is a bacterial zoonosis caused by pathogenic Leptospira for which rats are considered as the main reservoir. Disease incidence is higher in tropical countries, especially in insular ecosystems. Our objectives were to determine the current burden of leptospirosis in Seychelles, a country ranking first worldwide according to historical data, to establish epidemiological links between animal reservoirs and human disease, and to identify drivers of transmission. Methods A total of 223 patients with acute febrile symptoms of unknown origin were enrolled in a 12-months prospective study and tested for leptospirosis through real-time PCR, IgM ELISA and MAT. In addition, 739 rats trapped throughout the main island were investigated for Leptospira renal carriage. All molecularly confirmed positive samples were further genotyped. Results A total of 51 patients fulfilled the biological criteria of acute leptospirosis, corresponding to an annual incidence of 54.6 (95% CI 40.7–71.8) per 100,000 inhabitants. Leptospira carriage in Rattus spp. was overall low (7.7%) but dramatically higher in Rattus norvegicus (52.9%) than in Rattus rattus (4.4%). Leptospira interrogans was the only detected species in both humans and rats, and was represented by three distinct Sequence Types (STs). Two were novel STs identified in two thirds of acute human cases while noteworthily absent from rats. Conclusions This study shows that human leptospirosis still represents a heavy disease burden in Seychelles. Genotype data suggests that rats are actually not the main reservoir for human disease. We highlight a rather limited efficacy of preventive measures so far implemented in Seychelles. This could result from ineffective control measures of excreting animal populations, possibly due to a misidentification of the main contaminating reservoir(s). Altogether, presented data stimulate the exploration of alternative reservoir animal hosts., Author summary Leptospirosis is an emerging environmental infectious disease caused by corkscrew shaped bacteria called Leptospira. Humans usually get infected during recreational or work-related outdoor activities through contact with urine excreted by animal reservoirs. As a zoonotic disease, leptospirosis is a good example of the One Health concept for it links humans, animals and ecosystems in a web of pathogen maintenance and transmission. This zoonosis is highly prevalent in the tropics and especially in tropical islands. Seychelles archipelago has been reported as the country with highest human incidence worldwide, although figures are based on dated studies and/or poorly specific tests. The presented investigation aimed at providing an updated information on human leptospirosis burden in Seychelles and exploring the transmission chains in their environmental aspects. Presented data confirms that the disease still heavily impacts the country. Genotyping of pathogenic Leptospira in human acute cases reveals that three distinct Sequence Types (STs) are involved in the disease. However, rats typically considered as the main reservoir in Seychelles, harbor only one of these STs, found only in a minority of human cases. Hence, it appears that rats are likely not the main reservoir of leptospirosis in Seychelles, which has important consequences in terms of preventive measures to be implemented for a better control of human leptospirosis.

Background Although leptospirosis is a zoonosis of major concern on tropical islands, the molecular epidemiology of the disease aiming at linking human cases to specific animal reservoirs has been rarely explored within these peculiar ecosystems. Methodology/Principal Findings Five species of wild small mammals (n = 995) as well as domestic animals (n = 101) were screened for Leptospira infection on Reunion Island; positive samples were subsequently genotyped and compared to Leptospira from clinical cases diagnosed in 2012–2013 (n = 66), using MLST analysis. We identified two pathogenic species in human cases, namely Leptospira interrogans and Leptospira borgpetersenii. Leptospira interrogans was by far dominant both in clinical samples (96.6%) and in infected animal samples (95.8%), with Rattus spp and dogs being its exclusive carriers. The genetic diversity within L. interrogans was apparently limited to two sequence types (STs): ST02, identified among most clinical samples and in all rats with complete MLST, and ST34, identified in six humans, but not in rats. Noteworthy, L. interrogans detected in two stray dogs partially matched with ST02 and ST34. Leptospira borgpetersenii was identified in two clinical samples only (3.4%), as well as in cows and mice; four haplotypes were identified, of which two seemingly identical in clinical and animal samples. Leptospira borgpetersenii haplotypes detected in human cases were clearly distinct from the lineage detected so far in the endemic bat species Mormopterus francoismoutoui, thus excluding a role for this volant mammal in the local human epidemiology of the disease. Conclusions/Significance Our data confirm rats as a major reservoir of Leptospira on Reunion Island, but also pinpoint a possible role of dogs, cows and mice in the local epidemiology of human leptospirosis. This study shows that a comprehensive molecular characterization of pathogenic Leptospira in both clinical and animal samples helps to gaining insight into leptospirosis epidemiology within a specific environmental setting., Author Summary Leptospirosis is a zoonosis caused by infection with pathogenic Leptospira species. A broad range of animals, including rodents, pets and livestock, are maintenance hosts for leptospires. However, assessing the relative importance of each host in the contamination of the environment and, in fine, in the infection of humans, has rarely been performed. In this study, we surveyed various wild and domestic animal species and their Leptospira carriage in Reunion Island, where human leptospirosis is endemic. We determined and compared the Leptospira genetic diversity at the species and infra-species levels in laboratory-confirmed human cases and in infected animals. The two Leptospira species infecting humans, Leptospira interrogans and Leptospira borgpetersenii, could be traced back to different animal species: rats and dogs for the former species, cows and mice for the latter. The Leptospira infecting the single bat species endemic to the island was not found to be involved in human leptospirosis. Aside from rats, which were expected to play a role in the local epidemiology of the disease, the putative role of dogs, cattle and mice in human epidemiology on Reunion Island, pinpointed by our data, deserves a specific investigation. These results have strong implications in terms of local control actions aimed at reducing the burden of human leptospirosis.

Background Tick paralysis, resultant from envenomation by the scrub-tick Ixodes holocyclus, is a serious threat for small companion animals in the eastern coast of Australia. We hypothesise that surveillance systems that are built on Internet search queries may provide a more timely indication of high-risk periods more effectively than current approaches. Methods Monthly tick paralysis notifications in dogs and cats across Australia and the states of Queensland (QLD) and New South Wales (NSW) were retrieved from Disease WatchDog surveillance system for the period 2011–2013. Internet search terms related to tick paralysis in small companion animals were identified using Google Correlate, and corresponding search frequency metrics were downloaded from Google Trends. Spearman’s rank correlations and time series cross correlations were performed to assess which Google search terms lead or are synchronous with tick paralysis notifications. Results Metrics data were available for 24 relevant search terms at national level, 16 for QLD and 18 for NSW, and they were all significantly correlated with tick paralysis notifications (P

Globalization has facilitated the spread of numerous infectious agents to all corners of the planet. Analysis of the Global Infectious Disease and Epidemiology Network (GIDEON) database quantitatively illustrates that the globalization of human infectious agents depends significantly on the range of hosts used. Infectious agents specific to humans are broadly and uniformly distributed, whereas zoonotic infectious agents are far more localized in their geographical distribution. Moreover, these patterns vary depending on transmission mode and infectious agent taxonomy. This dichotomy is unlikely to persist if certain aspects of globalization (for example, exotic species introductions) continue unabated. This raises a serious concern for public health and leaves nations with the task of determining the infectious agents that have the greatest potential to establish within their borders. At the advent of a century characterized by an apparent increase in emerging infectious diseases, these results have critical implications for public-health policy and future research pathways of infectious disease ecology.

SUMMARYWe report herein the investigation of a leptospirosis outbreak occurring in triathlon competitors on Réunion Island, Indian Ocean. All participants were contacted by phone or email and answered a questionnaire. Detection and molecular characterization of pathogenicLeptospirawas conducted in inpatients and in rodents trapped at the vicinity of the event. Of the 160 athletes competing, 101 (63·1%) agreed to participate in the study. Leptospirosis was biologically confirmed for 9/10 suspected cases either by real-time PCR or serological tests (MAT or ELISA). The total attack rate, children's attack rate, swimmers’ attack rate, and the attack rate in adult swimmers were respectively estimated at 8·1% [95% confidence interval (CI) 4·3–14·7], 0%, 12·7% (95% CI 6·8–22·4) and 23·1% (95% CI 12·6–33·8). Leptospirosis cases reported significantly more wounds [risk ratio (RR) 4·5, 95% CI 1·6–13], wore complete neoprene suits less often (RR 4·3, 95% CI 1·3–14·5) and were most frequently unlicensed (RR 6·6, 95% CI 2·9–14·8). The epidemiological investigation supported that some measures such as the use of neoprene suits proved efficient in protecting swimmers against infection. PCR detection in rats revealed highLeptospirainfection rates. Partial sequencing of the 16S gene and serology on both human and animal samples strongly suggests that rats were the main contaminators and were likely at the origin of the infection in humans.

International audience; Connaître la distribution des espèces vectrices de zoonoses est fondamentale pour comprendre les dynamiques épidémiologiques et caractériser les zones à risque. Cette distribution peut être appréhendée à différentes échelles : régionalement, l'aire de répartition de ces espèces définit les limites d'occurrence de la transmission d'agents pathogènes et, localement, leurs niches déterminent l'aléa. Cette dernière échelle requiert une connaissance très précise des milieux qui peut être acquise par l'analyse d'images satellite à très haute résolution spatiale. Dans le cadre d'un projet de recherche sur la leptospirose à La Réunion (LeptOI), cette étude propose de mesurer le potentiel d'utilisation d'une image Pléiades (produit rééchantillonné à 50 cm) pour caractériser la distribution locale de différents petits mammifères terrestres autour de l'Etang de Saint-Paul. Trois campagnes de piégeage ont permis la capture de 134 animaux appartenant à quatre des cinq espèces présentes à La Réunion (Rattus rattus, R. norvegicus, Suncus murinus et Tenrec ecaudatus). Une analyse orientée-objet de l'image Pléiades a permis de discriminer dix classes d'occupation des sols avec une précision totale de 83,6%. Des indices paysagers ont été calculés à partir de cette classification autour de chacun des pièges (distances les plus courtes aux classes d'occupation, densité de contours et pourcentages de surface des classes au sein de zones tampon de 50 et de 100 mètres). Trois zones (« humide », « anthropisée » et « sèche ») sont clairement discriminées par ces variables, qui pourront être utilisées comme déterminants écologiques de différents profils épidémiologiques : la zone humide étant propice à la leptospirose et la zone anthropisée étant un lieu de cooccurrence des quatre espèces et ainsi potentiellement une zone d'échanges de pathogènes. Abstract Knowing the distribution of vectors of zoonoses is fundamental to understand epidemiological dynamics and characterize risk areas. This distribution can be assessed at different scales: regionally, the range of these species defines the limits of occurrence of pathogen transmission and, locally, niches determine the hazard. This latter scale requires a very precise knowledge of the areas that can be gained through the analysis of very high spatial resolution satellite images. As part of a research project on leptospirosis in La Réunion (LeptOI), this study proposes to measure the potential of a Pleiades image (product resampled to to 50 cm) to characterize the local distribution of various small terrestrial mammals in Saint-Paul swamp area. Three trapping campaigns have led to the capture of 134 animals belonging to four of the five species occurring in Reunion Island (Rattus rattus, R. norvegicus, Suncus murinus and Tenrec ecaudatus). An object-based image analysis of the Pleiades image allowed discriminating ten land use classes with a total accuracy of 83.6%. Landscape indices were calculated from the classification around each trap (shortest distances to each land use class, edge densities and proportion of each patch within buffers defined by a 50 meters or a 100 meters radius). Three areas ("wet", "urbanized" and "dry") are clearly discriminated by these variables, which can be used as ecological determinants of different epidemiological profiles. The wetland area is known as suitable to leptospirosis whereas the urbanized area is a place of co-occurrence of the four species and thus potentially favorable to the exchange of pathogens.

The three French overseas departments of the Americas are characterized both by insular (Guadeloupe and Martinique) and continental (French Guiana) settings with a tuberculosis case detection rate that varies from less than 10 per 100,000 per year in insular areas to an estimated incidence of more than 55 per 100,000 in French Guiana. Under a long-term genotyping program, more than three-fourths of all the Mycobacterium tuberculosis isolates ( n = 744) received from the three settings were fingerprinted over a 10-year period (1994 to 2003) by spoligotyping and variable number of tandem DNA repeats (VNTRs) in order to understand the current trends in their detection rates, drug resistance, and groups and subpopulations at risk of contracting the disease and to pinpoint the circulating phylogeographical clades of the bacilli. The major difference in the study populations was the nationality of the patients, with a high percentage of immigrants from high-incidence neighboring countries in French Guiana and a low but increasing percentage in the French Caribbean. The rate of recent transmission was calculated to be 49.3% in French Guiana, compared to 27.2% and 16.9% in Guadeloupe and Martinique, respectively. At the phylogeographic level, 77.9% of the isolates studied belonged to four major clades (Haarlem, Latin-American and Mediterranean, T, and X) which are already reported from neighboring Caribbean islands in an international database and may underline potential interregional transmission events.

SummaryThe human leukocyte antigen (HLA; known as MHC in other vertebrates) plays a central role in the recognition and presentation of antigens to the immune system and represents the most polymorphic gene cluster in the human genome [1]. Pathogen-driven balancing selection (PDBS) has been previously hypothesized to explain the remarkable polymorphism in the HLA complex, but there is, as yet, no direct support for this hypothesis [2, 3]. A straightforward prediction coming out of the PDBS hypothesis is that populations from areas with high pathogen diversity should have increased HLA diversity in relation to their average genomic diversity. We tested this prediction by using HLA class I genetic diversity from 61 human populations. Our results show that human colonization history explains a substantial proportion of HLA genetic diversity worldwide. However, between-population variation at the HLA class I genes is also positively correlated with local pathogen richness (notably for the HLA B gene), thus providing support for the PDBS hypothesis. The proportion of variations explained by pathogen richness is higher for the HLA B gene than for the HLA A and HLA C genes. This is in good agreement with both previous immunological and genetic data suggesting that HLA B could be under a higher selective pressure from pathogens.

The diversity and geographical distribution of fleas parasitizing small mammals have been poorly investigated on Indian Ocean islands with the exception of Madagascar where endemic plague has stimulated extensive research on these arthropod vectors. In the context of an emerging flea-borne murine typhus outbreak that occurred recently in Reunion Island, we explored fleas' diversity, distribution and host specificity on Reunion Island. Small mammal hosts belonging to five introduced species were trapped from November 2012 to November 2013 along two altitudinal transects, one on the windward eastern and one on the leeward western sides of the island. A total of 960 animals were trapped, and 286 fleas were morphologically and molecularly identified. Four species were reported: (i) two cosmopolitan Xenopsylla species which appeared by far as the prominent species, X. cheopis and X. brasiliensis; (ii) fewer fleas belonging to Echidnophaga gallinacea and Leptopsylla segnis. Rattus rattus was found to be the most abundant host species in our sample, and also the most parasitized host, predominantly by X. cheopis. A marked decrease in flea abundance was observed during the cool-dry season, which indicates seasonal fluctuation in infestation. Importantly, our data reveal that flea abundance was strongly biased on the island, with 81% of all collected fleas coming from the western dry side and no Xenopsylla flea collected on almost four hundred rodents trapped along the windward humid eastern side. The possible consequences of this sharp spatio-temporal pattern are discussed in terms of flea-borne disease risks in Reunion Island, particularly with regard to plague and the currently emerging murine typhus outbreak., Author Summary Fleas are blood-feeding parasites involved in the transmission of several arthropod borne pathogens. Rat-fleas (Xenopsylla spp.) are known vectors of bubonic plague together with other human diseases receiving less attention such as murine typhus. This latter disease was recorded for the first time in 2011 on Reunion Island where seven human cases were further confirmed within the following year. The outbreak motivated a large survey of fleas, as these insects of major veterinary and medical importance have never been investigated on this oceanic island. We collected fleas on almost 1000 small wild mammals trapped on two altitudinal transects along the humid eastern and dry western sides of the island. Our data reveal the presence of four cosmopolitan flea species and shows an astonishing distribution pattern: 81% of all collected fleas were sampled on the western transect while not a single rat-flea was sampled on the eastern humid side of the island. Interestingly, this distribution did at least in part overlay the map of murine typhus human cases. These data stimulate the need for a diagnosis of pathogens in natural flea populations together with a comprehensive distribution map of fleas, allowing a risk assessment of flea-borne diseases in humans.

The spatial dynamics of zoonotic arthropod-borne viruses is a fashionable though challenging topic. Inter-human local transmission of a given arbovirus during an outbreak and its spread over large distances are considered as key parameters of emergence. Here, we suggest that insular ecosystems provide ideal natural ''laboratory'' conditions to uncouple local transmission from long distance spread, and differentiate these two processes. Due to geographic isolation, often-limited land surface area and relatively homogenous ecosystems, oceanic islands display low species richness and often-high levels of endemism. These aspects provide the means for comprehensive entomological surveys and investigations of original host/pathogen interactions. In addition, islands are interconnected through discrete anthropogenic and non-anthropogenic exchanges: whilst islands maintain a substantial level of human and domestic animal exchange with other neighbouring or distant territories, they also comprise dispersal and migratory pathways of volant organisms (insects, birds and bats). Hence, both anthropogenic and non-anthropogenic exchanges in island systems are easier to identify and investigate than in continuous, continental systems. Finally, island ecosystems tend to be notably simpler, more prone to invasive taxa and, therefore, easier to document the colonization or displacement of vector species. These different aspects are presented and overlaid upon the spread of arboviruses within two distinct insular systems: islands of Polynesia and the south-western Indian Ocean. The former have been repeatedly affected by Dengue fever epidemics, while the latter recently suffered four successive epidemics, probably of east African origin, three of which involved the emerging viruses Chikungunya, Rift Valley and Dengue fever. Here, we review some new insights into arboviral spread and evolution associated with investigations that followed these epidemics, as well as several aspects that make insular ecosystems favourable to the investigation of arboviral transmission and spread.

To the Editor: Leptospirosis is a zoonosis of global distribution; incidence rates are particularly high in tropical areas (1). Leptospirosis is a major public health problem on islands in the southwestern Indian Ocean, particularly La Reunion, Mayotte, and the Seychelles (where incidence rates are among the highest in the world) (1). In contrast, no human case has been reported on the nearby islands of Madagascar and Union of the Comoros. However, the recent demonstration of pathogenic Leptospira spp. in small mammals introduced to Madagascar suggests possible transmission from free-living animals to humans (2). In addition to the fact that incidence rates vary among humans, clinical bacterial isolates from different islands belong to different serogroups and serovars and show diverse molecular features (3,4). This diversity might be correlated with that of the reservoir hosts; the islands in the southwestern Indian Ocean are a hot spot of biodiversity with extraordinary levels of vertebrate endemism. Most studies investigating wild-animal reservoirs of Leptospira spp. on the islands in the southwestern Indian Ocean have focused on small mammals that had been introduced to the islands (2,5), although bats infected with pathogenic Leptospira spp. have been identified in other regions (6). Whether bats are a reservoir of Leptospira spp. on these islands remains unknown. Therefore, we looked for this bacterium in bats from Madagascar and Union of the Comoros and characterized associated genetic diversity. As part of an ongoing program aimed at identifying viral and bacterial infectious agents in island wild fauna, 129 insectivorous and frugivorous bats were tested for Leptospira spp. The bats belonged to 12 species: 9 from Madagascar (Mormopterus jugularis, Otomops madagascariensis, Triaenops furculus, T. menamena, Miniopterus gleni, Miniopterus griffithsi, Miniopterus mahafaliensis, Myotis goudoti, Hypsugo anchietae) and 3 from Union of the Comoros (Rousettus obliviosus, Chaerephon pusillus, Miniopterus griveaudi). Bats were captured in mist nets or harp traps at 8 sites in Madagascar and 6 sites in Union of the Comoros. Organs were collected in the field and immediately stored in liquid nitrogen. Total nucleic acids were extracted from a pool of kidney, spleen, and lung tissue by using the Biorobot EZ1 and EZI Virus Mini Kit version 2.0 (QIAGEN, Les Ulis, France). Reverse transcription was then performed with GoScript reverse transcriptase (Promega, Charbonnieres-les-Bains, France) to obtain cDNA. We screened pathogenic Leptospira spp. with a probe-specific real-time PCR (7). The 25 positive samples were subsequently subjected to a PCR procedure that amplified fragments from 682 to 1,293 bp of the 16S rRNA gene (depending on the amplification success) by using published primers (8–10). Resulting PCR products from 7 samples were sequenced and compared with available sequences in GenBank by using phylogenetic construction with PhyML 3.0. Of the 12 bat species tested, 11 were positive for Leptospira spp. (the only H. anchietae bat tested was negative). Among 52 bats from Madagascar, 18 (34.6%) were infected; detection rates were often high, e.g., 8 (80%) of 10 T. menamena bats. In contrast, among 77 bats from Union of the Comoros, only 9 (11.7%) were infected. Leptospira spp. seem to be ubiquitous in the study areas; infected bats were found at 7 of 8 sites in Madagascar and 3 of 6 sites in the Union of the Comoros. Of the 7 Leptospira spp. sequences obtained from bats in this study, 3 were closely related to L. borgpetersenii, 1 grouped with L. interrogans, and 3 were not associated with any described species (Figure). L. interrogans and L. borgpetersenii were identified from R. obliviosus bats from the same cave in the Union of the Comoros, and the L. borgpetersenii sequence was closely related to that identified from the O. madagascariensis bats, which are endemic to Madagascar. Potentially pathogenic Leptospira spp. were found in bats of a wide variety of species in Madagascar and Union of the Comoros, at most study sites, and at levels notably higher than those reported from similar studies in other regions (6). Some of the bats that were Leptospira spp.–positive by PCR, particularly the genera Mormopterus and Chaerephon, often occupy synanthropic day roost sites. For example, we sampled 1 positive colony of C. pusillus bats in a school attic (Pomoni, Anjouan, Union of the Comoros), and bat scats were visible on the floor within the classroom. Figure Maximum-likelihood phylogenetic tree of Leptospira spp.16s rDNA in bats from Madagascar and the Union of the Comoros. The dendrogram was constructed with a fragment of 641 bp, with the TIMef+I+G substitution model, and with 1,000 replications. Only bootstrap ... Bats from Madagascar and Union of the Comoros harbor a notable diversity of Leptospira spp.; this finding is in accordance with the diversity found in a comparable investigation of bats in the Amazon region (6). Although leptospirosis in humans is suspected only on the islands associated with this study (10), incidence among humans in Mayotte, part of the Union of the Comoros archipelago, has been shown to be high and mainly associated with L. borgpetersenii (3). The use of more polymorphic markers combined with the sequencing of clinical isolates should provide better characterization of Leptospira spp. diversity and the potential role of bats in human leptospirosis.

International audience; BACKGROUND: To date, there is little information that reflects the true extent of spread of the pH1N1/2009v influenza pandemic at the community level as infection often results in mild or no clinical symptoms. This study aimed at assessing through a prospective study, the attack rate of pH1N1/2009 virus in Reunion Island and risk factors of infection, during the 2009 season. METHODOLOGY/PRINCIPAL FINDINGS: A serosurvey was conducted during the 2009 austral winter, in the frame of a prospective population study. Pairs of sera were collected from 1687 individuals belonging to 772 households, during and after passage of the pandemic wave. Antibodies to pH1N1/2009v were titered using the hemagglutination inhibition assay (HIA) with titers ≥ 1/40 being considered positive. Seroprevalence during the first two weeks of detection of pH1N1/2009v in Reunion Island was 29.8% in people under 20 years of age, 35.6% in adults (20-59 years) and 73.3% in the elderly (≥ 60 years) (P

International audience; Many debates surround the generalization of Rapoport's rule (i.e., the presence of a positive correlation between range size and latitude); however, little attention has been devoted to this spatial pattern (1) worldwide and (2) for pathogenic microorganisms. In this study, we analyzed this relationship for 290 human pathogenic species dispersed throughout the world to test whether pathogenic organisms with different ecological niches and strategies will show this trend. The midpoint method was used to calculate the correlation between the geographical range size and the latitude applied to different subsets of pathogens, including taxonomic subdivisions (bacteria, viruses, helminths, protozoans, and fungi) and categories based on transmission mode and host specificity. It is assumed that Rapoport's spatial pattern may exist for human infectious diseases, whatever hemisphere is considered, for 5 to 7 of 8 of the selected groups, depending on the pathogen species included. This is the first study performed to investigate Rapoport's pattern at a global scale for various pathogenic organisms. We also discuss how three well-known spatial patterns of diversity, i.e., latitudinal gradient, nested species pattern, and Rapoport's rule, may vary together to produce the actual large-scale geographical distribution of human pathogenic species observed on Earth. These findings have important messages for understanding the distribution and the diffusion of human pathogenic species; however, further studies are needed to investigate the exact underlying mechanisms responsible for those patterns.

Background French Guiana has the highest tuberculosis (TB) burden among all French departments, with a strong increase in the TB incidence over the last few years. It is now uncertain how best to explain this incidence. The objective of this study was to compare three different methods evaluating the extent of recent TB transmission in French Guiana. Methods We conducted a population-based molecular epidemiology study of tuberculosis in French Guiana based on culture-positive TB strains (1996 to 2003, n = 344) to define molecular relatedness between isolates, i.e. potential transmission events. Phylogenetic relationships were inferred by comparing two methods: a "cluster-graph" method based on spoligotyping results, and a minimum spanning tree method based on both spoligotyping and variable number of tandem DNA repeats (VNTR). Furthermore, three indices attempting to reflect the extent of recent TB transmission (RTIn, RTIn-1 and TMI) were compared. Results Molecular analyses showed a total amount of 120 different spoligotyping patterns and 273 clinical isolates (79.4%) that were grouped in 49 clusters. The comparison of spoligotypes from French Guiana with an international spoligotype database (SpolDB4) showed that the majority of isolates belonged to major clades of M. tuberculosis (Haarlem, 22.6%; Latin American-Mediterranean, 23.3%; and T, 32.6%). Indices designed to quantify transmission of tuberculosis gave the following values: RTIn = 0.794, RTIn-1 = 0.651, and TMI = 0.146. Conclusion Our data showed a high number of Mycobacterium tuberculosis clusters, suggesting a high level of recent TB transmission, nonetheless an estimation of transmission rate taking into account cluster size and mutation rate of genetic markers showed a low ongoing transmission rate (14.6%). Our results indicate an endemic mode of TB transmission in French Guiana, with both resurgence of old spatially restricted genotypes, and a significant importation of new TB genotypes by migration of TB infected persons from neighgouring high-incidence countries.