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Introduction: Widespread transmission of Japanese encephalitis virus (JEV) genotype four (GIV) occurred across mainland Australia in 2022. This resulted in forty-five human cases, including seven deaths, and the identification of JEV infection in over 80 commercial piggeries. Materials and Methods: We collected mosquitoes which were trapped using CO 2 -baited light traps deployed near piggeries reporting disease or in regions linked to human cases in the Wide Bay region in the state of Queensland. Mosquitoes from four traps yielded JEV RNA by real-time RT-PCR. Pools containing RNA positive mosquitoes were inoculated onto mosquito cell monolayers. Discussion: A single isolate of JEV was obtained from a pool of mixed mosquito species. Near whole genome sequencing and phylogenetic analysis of the JEV isolate demonstrated its high genomic relatedness with JEV GIV pig sequences sampled from Queensland and the state of New South Wales in 2022. Conclusion: We report the first isolation of JEV GIV from mosquitoes collected in Australia. With only a few JEV GIV isolates available globally, the isolate we report will be essential for future research of JEV host interactions, evolution and disease markers, and development of effective therapies, vaccines, diagnostic assays, and mosquito control strategies.

The study of counterproductive work behavior (CWB), intentional actions by employees that are deleterious to the organization and/or its stakeholders, has produced research on the dimensionality of CWB, as well as its situational and dispositional antecedents. Absent from these advancements have been investigations into the potential utility of a taxonomy of counterproductive employee types-a "person-oriented" approach. Our latent profile analysis ( N = 522) suggested a four-profile solution which included one profile with uniformly low rates across CWBs (here termed "Angels;" 14% of the sample), and three profiles with higher CWB rates but which were distinguishable by different CWBs being most frequent in each group. Specifically, one profile was distinguished from the Angels group by higher rates of less severe CWBs (misuse of time/resources and poor attendance; 33% of the sample). The other two of the three counterproductive profiles were similar to each other except that one was characterized by higher drug use than the other (14% of the sample). The profiles also differed significantly on narcissism, psychopathy, and Machiavellianism, and on self-reports of prior arrest and censure by employers. Provided these distinctions among profiles, the treatment and assumptions of employee counterproductivity in research and practice should be revisited, particularly when using models assuming a homogenous, monotonic relationship between counterproductive behaviors across employees. Implications for our conceptual understanding of counterproductivity and applied interventions aimed at reducing CWBs are discussed, alongside recommendations for future person-oriented research on CWB., Competing Interests: The authors declare that they have no conflict of interests., (© Person-Oriented Research.)

Purpose: The clinical spectrum of motile ciliopathies includes laterality defects, hydrocephalus, and infertility as well as primary ciliary dyskinesia when impaired mucociliary clearance results in otosinopulmonary disease. Importantly, approximately 30% of patients with primary ciliary dyskinesia lack a genetic diagnosis., Methods: Clinical, genomic, biochemical, and functional studies were performed alongside in vivo modeling of DAW1 variants., Results: In this study, we identified biallelic DAW1 variants associated with laterality defects and respiratory symptoms compatible with motile cilia dysfunction. In early mouse embryos, we showed that Daw1 expression is limited to distal, motile ciliated cells of the node, consistent with a role in left-right patterning. daw1 mutant zebrafish exhibited reduced cilia motility and left-right patterning defects, including cardiac looping abnormalities. Importantly, these defects were rescued by wild-type, but not mutant daw1, gene expression. In addition, pathogenic DAW1 missense variants displayed reduced protein stability, whereas DAW1 loss-of-function was associated with distal type 2 outer dynein arm assembly defects involving axonemal respiratory cilia proteins, explaining the reduced cilia-induced fluid flow in particle tracking velocimetry experiments., Conclusion: Our data define biallelic DAW1 variants as a cause of human motile ciliopathy and determine that the disease mechanism involves motile cilia dysfunction, explaining the ciliary beating defects observed in affected individuals., Competing Interests: Conflict of Interest The authors declare no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Motile cilia generate cell propulsion and extracellular fluid flows that are crucial for airway clearance, fertility and left-right patterning. Motility is powered by dynein arm complexes that are assembled in the cytoplasm then imported into the cilium. Studies in Chlamydomonas reinhardtii showed that ODA16 is a cofactor which promotes dynein arm import. Here, we demonstrate that the zebrafish homolog of ODA16, Daw1, facilitates the onset of robust cilia motility during development. Without Daw1, cilia showed markedly reduced motility during early development; however, motility subsequently increased to attain close to wild-type levels. Delayed motility onset led to differential effects on early and late cilia-dependent processes. Remarkably, abnormal body axis curves, which formed during the first day of development due to reduced cilia motility, self-corrected when motility later reached wild-type levels. Zebrafish larva therefore possess the ability to survey and correct body shape abnormalities. This work defines Daw1 as a factor which promotes the onset of timely cilia motility and can explain why human patients harboring DAW1 mutations exhibit significant laterality perturbations but mild airway and fertility complications., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Background: Leptospirosis is a zoonotic disease with a worldwide distribution, caused by pathogenic serovars in the genus Leptospira. Feral pigs are known carriers of Leptospira species and pig hunting using dogs is a common recreational activity in Queensland, Australia., Methodology and Principal Findings: This study aimed to determine the seroprevalence of Leptospira spp. serovars in pig-hunting dogs above the Tropic of Capricorn in Queensland and by establishing the geographic distribution, serovars and incidence of human cases of leptospirosis in Queensland, identify potential overlap between human and canine exposure. We also explored the knowledge and risk-taking behaviours of pig-hunting dog owners towards zoonotic diseases. Ninety-eight pig-hunting dogs deemed healthy by physical examination and owned by 41 people from Queensland had serum submitted for Microscopic Agglutination Testing (MAT) to determine antibody titres against Leptospira serovars, while 40/41 dog owners completed a survey on their knowledge of diseases relating to pig hunting. Human leptospirosis cases (n = 330) notified to Queensland Health between 2015-2018 were analysed. Approximately one quarter (23/87; 26%) of unvaccinated pig-hunting dogs were seropositive to Leptospira spp. Although harder to interpret, 8/11 (73%) vaccinated dogs were seropositive to Leptospira spp. Pig hunters may be more likely to contract leptospirosis compared with the general Queensland population, based on responses from surveyed hunters. The highest concentration of human leptospirosis was in the wet tropics region of Far North Queensland. There was little overlap between the serovars dogs were exposed to and those infecting humans. The dominant serovar identified in unvaccinated dogs was Australis (13/23; 57%), with serovar Arborea (36/330; 10.9%) responsible for the highest number of human leptospirosis cases. Topaz was the second most common serovar in both humans and dogs and was previously unrecorded in Australian dogs. Most hunters surveyed used hand washing as a zoonotic disease risk reduction technique., Conclusions: Leptospirosis is an emerging disease of growing significance. The infection requires a 'one health' approach to understand its epidemiology. With shifting climatic patterns influencing human-animal-environment interactions, ongoing monitoring of diseases like leptospirosis is critical to helping prevent infection of individuals and disease outbreaks., Competing Interests: The authors have declared that no competing interests exist.

Traditional mouse models of lyssavirus pathogenesis rely on euthanizing large groups of animals at various time points post-infection, processing infected tissues, and performing histological and molecular analyses to determine anatomical sites of infection. While powerful by some measures, this approach is limited by the inability to monitor disease progression in the same mice over time. In this study, we established a novel non-invasive mouse model of lyssavirus pathogenesis, which consists of longitudinal imaging of a luciferase-expressing Australian bat lyssavirus (ABLV) reporter virus. In vivo bioluminescence imaging (BLI) in mice revealed viral spread from a peripheral site of inoculation into the central nervous system (CNS), with kinetically and spatially distinct foci of replication in the footpad, spinal cord, and hindbrain. Detection of virus within the CNS was associated with onset of clinical disease. Quantification of virus-derived luminescent signal in the brain was found to be a reliable measure of viral replication, when compared to traditional molecular methods. Furthermore, we demonstrate that in vivo imaging of ABLV infection is not restricted to the use of albino strains of mice, but rather strong BLI signal output can be achieved by shaving the hair from the heads and spines of pigmented strains, such as C57BL/6. Overall, our data show that in vivo BLI can be used to rapidly and non-invasively identify sites of lyssavirus replication and to semi-quantitatively determine viral load without the need to sacrifice mice at multiple time points., Competing Interests: Declaration of Competing Interest None., (Published by Elsevier B.V.)

Background: In 2018, an outbreak of leptospirosis was identified among raspberry workers from a mixed-berry farm in New South Wales, Australia. Initial testing had not revealed a cause, but eventually leptospirosis was detected via polymerase chain reaction (PCR). Further serological testing detected Leptospira borgpetersenii serovar Arborea, of which rodents are the predominant reservoir. Leptospirosis is rare in Australia, with outbreaks usually related to flooding. We conducted an investigation to identify risk factors for infection, to inform control measures., Methods: Cases were detected through laboratory notifications, hospital-based syndromic surveillance, awareness-raising among farm employees and clinician alerts. Confirmed cases had a four-fold rise in antibody titre or single titre ≥400 on microscopic agglutination test, and a positive IgM. Probable cases had a positive Leptospira PCR or IgM, and possible cases had a clinically compatible illness. We conducted a case-control study among raspberry workers on the farm and compared reported exposures between cases and seronegative controls. We assessed environmental risks on-site and tested rodents for leptospirosis., Results: We identified 84 cases over a 5-month period (50 confirmed, 19 probable and 15 possible). Compared with controls, cases were less likely to wear gloves and more recently employed. Cases also more commonly reported always having scratched hands, likely from the thorns on raspberry plants. We observed evidence of rodent activity around raspberry plants and three of thirteen trapped mice tested positive for Leptospira Arborea. Control measures included enhanced glove use, doxycycline prophylaxis and rodent control., Conclusions: This is the largest known outbreak of leptospirosis in Australia. Workers were likely exposed through scratches inflicted during harvesting, which became contaminated with environmental leptospires from mice. Leptospirosis should be considered an occupational risk for raspberry workers, requiring protective measures. Chemoprophylaxis may assist in controlling outbreaks. PCR assists in the early diagnosis and detection of leptospirosis and should be included in surveillance case definitions., (© 2019 Blackwell Verlag GmbH.)

During 2003-2011, we recruited 1,065 patients of all ages admitted to Mahosot Hospital (Vientiane, Laos) with suspected central nervous system (CNS) infection. Etiologies were laboratory confirmed for 42.3% of patients, who mostly had infections with emerging pathogens: viruses in 16.2% (mainly Japanese encephalitis virus [8.8%]); bacteria in 16.4% (including Orientia tsutsugamushi [2.9%], Leptospira spp. [2.3%], and Rickettsia spp. [2.3%]); and Cryptococcus spp. fungi in 6.6%. We observed no significant differences in distribution of clinical encephalitis and meningitis by bacterial or viral etiology. However, patients with bacterial CNS infection were more likely to have a history of diabetes than others. Death (26.3%) was associated with low Glasgow Coma Scale score, and the mortality rate was higher for patients with bacterial than viral infections. No clinical or laboratory variables could guide antibiotic selection. We conclude that high-dependency units and first-line treatment with ceftriaxone and doxycycline for suspected CNS infections could improve patient survival in Laos.

Leptospirosis is an important zoonotic disease in Nepal; however, there is a lack of information on sources of leptospirosis infection for people and associated risk factors. We implemented a case-control study nested within a cross-sectional survey to investigate zoonotic risks of human leptospirosis among adult, febrile patients seeking treatment in healthcare centres in Kaski District, Nepal, from April to October 2013. The study population was 239 febrile patients; the data consisted of paired blood samples; questionnaires on clinical symptoms; occupational, environmental and animal exposures; and a blood sample from animals in the household. In total, 63 cattle, 92 buffalo, 181 goats, 20 dogs and 14 rodents from 119 households were blood sampled. Serology was performed on the samples using the microscopic agglutination test (MAT) for a panel of 20 serovars with a starting dilution of 1:50. Clinical leptospirosis was defined as a titre of 1:400 or stronger, or a 4-fold or greater rise between acute and convalescent titres, or evidence of seroconversion for one or more serovars represented by a titre of <1:50 in the acute sample and a titre of ≥1:100 in the convalescent sample. The prevalence of clinical leptospirosis was 5.4% (95% CI 2.6%-8.3%). The most common symptoms among recruited participants were loss of appetite and lethargy followed by chills, profuse sweating and backache. No individual clinical symptom nor combination of any two symptoms was significantly associated with clinical leptospirosis. However, owning goats (OR 1.3, CI 95% 1.05-1.66), working in rice fields (OR 1.3, CI 95% 1.11-1.72) and male gender (OR 4, CI 95% 1.12-17.26) significantly increased the risk of clinical leptospirosis in humans. Findings suggest that leptospirosis should be considered in the clinical differential diagnosis among febrile adult patients in Nepal, especially for men, goat owners and/or those who work in rice fields., (© 2018 Blackwell Verlag GmbH.)

Severe flooding has been linked to outbreaks of leptospirosis. Two sequential extreme flood events in Western Fiji caused the largest outbreak of leptospirosis recorded in the South Pacific, with 1,217 total suspected cases, of which 314 were probable and confirmed. Most (83%) cases occurred within 6 weeks of the flood events, displaying a biphasic epidemic curve associated with the floods. Given the temporal proximity of cases to flooding events, most of the transmission appeared to occur during or immediately after the floods; therefore, prevention of exposure to contaminated environments is a priority in the immediate flood and post-flood period. In addition, genotyping studies suggest that multiple animal reservoirs were implicated in the outbreak, reaffirming the importance of integrated human and animal health strategies for leptospirosis control.

Objectives: To compare two molecular assays (rrs quantitative PCR (qPCR) versus a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos., Methods: Serum, buffy coat and urine samples were collected on admission, and follow-up serum ∼10 days later. Leptospira spp. culture and microscopic agglutination tests (MAT) were performed as reference standards. Bayesian latent class modelling was performed to estimate sensitivity and specificity of each diagnostic test., Results: In all, 787 patients were included in the analysis: 4/787 (0.5%) were Leptospira culture positive, 30/787 (3.8%) were MAT positive, 76/787 (9.7%) were rrs qPCR positive and 20/787 (2.5%) were 16SrRNA/LipL32 qPCR positive for pathogenic Leptospira spp. in at least one sample. Estimated sensitivity and specificity (with 95% CI) of 16SrRNA/LipL32 qPCR on serum (53.9% (33.3%-81.8%); 99.6% (99.2%-100%)), buffy coat (58.8% (34.4%-90.9%); 99.9% (99.6%-100%)) and urine samples (45.0% (27.0%-66.7%); 99.6% (99.3%-100%)) were comparable with those of rrs qPCR, except specificity of 16SrRNA/LipL32 qPCR on urine samples was significantly higher (99.6% (99.3%-100%) vs. 92.5% (92.3%-92.8%), p <0.001). Sensitivities of MAT (16% (95% CI 6.3%-29.4%)) and culture (25% (95% CI 13.3%-44.4%)) were low. Mean positive Cq values showed that buffy coat samples were more frequently inhibitory to qPCR than either serum or urine (p <0.001)., Conclusions: Serum and urine are better samples for qPCR than buffy coat, and 16SrRNA/LipL32 qPCR performs better than rrs qPCR on urine. Quantitative PCR on admission is a reliable rapid diagnostic tool, performing better than MAT or culture, with significant implications for clinical and epidemiological investigations of this global neglected disease., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Leptospirosis is a globally important cause of acute febrile illness, and a common cause of non-malarial fever in Asia, Africa, and Latin America. Simple rapid diagnostic tests (RDTs) are needed to enable health-care workers, particularly in low resource settings, to diagnose leptospirosis early and give timely targeted treatment. This study compared four commercially available RDTs to detect human IgM against Leptospira spp. in a head-to-head prospective evaluation in Mahosot Hospital, Lao PDR. Patients with an acute febrile illness consistent with leptospirosis ( N = 695) were included in the study during the 2014 rainy season. Samples were tested with four RDTs: ("Test-it" [Life Assay, Cape Town, South Africa; N = 418]; "Leptorapide" [Linnodee, Ballyclare, Northern Ireland; N = 492]; "Dual Path Platform" [DPP] [Chembio, Medford, NY; N = 530]; and "SD-IgM" [Standard Diagnostics, Yongin, South Korea; N = 481]). Diagnostic performance characteristics were calculated and compared with a composite reference standard combining polymerase chain reaction (PCR) ( rrs ), microscopic agglutination tests (MATs), and culture. Of all patients investigated, 39/695 (5.6%) were positive by culture, PCR, or MAT. The sensitivity and specificity of the RDTs ranged greatly from 17.9% to 63.6% and 62.1% to 96.8%, respectively. None of the investigated RDTs reached a sensitivity or specificity of > 90% for detecting Leptospira infections on admission. In conclusion, our investigation highlights the challenges associated with Leptospira diagnostics, particularly in populations with multiple exposures. These findings emphasize the need for extensive prospective evaluations in multiple endemic settings to establish the value of rapid tools for diagnosing fevers to allow targeting of antibiotics.

Background: Leptospirosis is one of the most common zoonotic diseases worldwide. Infection occurs through contact with infected animals, or soil or water that has been contaminated by the urine of infected animals. Risk factors include occupational and recreational exposures, contact with floodwaters, and travel to areas with a high risk of leptospirosis, particularly tropical, developing countries. With climate change, flood-related outbreaks are becoming more common., Objective: This article aims to improve awareness of leptospirosis, and provide an update for general practitioners on its epidemiology, risk factors, clinical presentation, laboratory diagnosis, management and prevention., Discussion: Leptospirosis is sometimes misdiagnosed because clinical presentation can be non-specific and overlap with many other causes of acute febrile illnesses. In patients with risk factors for leptospirosis, a high index of clinical suspicion is important to ensure early diagnosis and treatment. Delays in treatment could increase the risk of severe complications, including pulmonary haemorrhage, acute renal failure and acute liver failure.

In 2014, we performed a diagnostic study of leptospirosis in Tasmanian devil ( Sarcophilus harrisii ) samples collected between 2008 and 2012 from wild and captive animals. Tasmanian devil populations have been declining because of a facial tumor disease since the 1990s, with ongoing investigations examining potential causative agents. Identifying other causative pathogens that may contribute additively to their decline is important to preserve current and future populations. We tested 81 Tasmanian devil serum samples and two tissue samples using PCR, microscopic agglutination test (MAT), and microsphere immunoassay (MIA). We found evidence of leptospirosis in Tasmanian devil populations across a wide geographic range of Tasmania. Antibodies to serovars in the serogroup Javanica, which are not considered endemic to Australia, were identified in 10 Tasmanian devils using MAT. We also identified serovar Celledoni serologically using the immunoglobulin G MIA and detected Leptospira in one sample using PCR.

Introduction: Laos has the highest maternal mortality ratio in mainland Southeast Asia and a high incidence of infectious diseases. Globally, malaria has been the pathogen most intensively investigated in relation to impact on pregnancy, but there has been relatively little research on the aetiology and impact of other diseases. We therefore aimed to determine the causes and impact of fever in pregnant women admitted to two central hospitals in Vientiane City, Lao PDR (Laos)., Materials and Methods: This hospital-based prospective study was conducted in Mahosot Hospital and the Mother and Child Hospital, Vientiane, between 2006 and 2010, with the aim to recruit 250 consenting pregnant women admitted with tympanic temperature ≥37.5°C. Primary outcome was the cause of fever and secondary outcomes were pregnancy outcomes. Specific investigations (culture, antigen, molecular and serological tests) were performed to investigate causes of fever. After discharge, all pregnant women were asked to return for review and convalescence serum on day 10-14 and were monitored until delivery., Principle Findings: 250 pregnant women were recruited to this study between February 2006 and November 2010. Fifty percent were pregnant for the first time. Their median (range) gestational age on admission was 24 (4-43) weeks. The median (range) tympanic admission temperature was 38.5°C (37.5-40.5°C). Fifteen percent of patients stated that they had taken antibiotics before admission. Headache, myalgia, back pain and arthralgia were described by >60% of patients and 149 (60%) were given a laboratory diagnosis. Of those with confirmed diagnoses, 132 (53%) had a single disease and 17 (7%) had apparent mixed diseases. Among those who had a single disease, dengue fever was the most common diagnosis, followed by pyelonephritis, scrub typhus, murine typhus and typhoid. Patients were also diagnosed with tuberculosis, appendicitis, Staphylococcus aureus septicemia, leptospirosis, Japanese encephalitis virus infection and Plasmodium falciparum malaria. Severe consequences, including maternal death, miscarriage, stillbirth, low birth weight and preterm birth, were found among 28 (78%) mothers with dengue fever, rickettsioses and typhoid., Conclusion: Fevers other than malaria, such as dengue, pyelonephritis, rickettsioses and typhoid are common causes of fever during pregnancy in the Asian tropics. Further investigations of their impact in the community on maternal death, fetal loss, vertical transmission, low birth weight and preterm birth are needed.

Leptospirosis is an important zoonosis worldwide, with infections occurring after exposure to contaminated water. Despite being a global problem, laboratory diagnosis remains difficult with culture results taking up to 3 months, serology being retrospective by nature, and polymerase chain reaction showing limited sensitivity. Leptospira have been shown to survive and multiply in blood culture media, and we hypothesized that extracting DNA from incubated blood culture fluid (BCF), followed by quantitative real-time polymerase chain reaction (qPCR) could improve the accuracy and speed of leptospira diagnosis. We assessed this retrospectively, using preincubated BCF of Leptospira spp. positive (N= 109) and negative (N= 63) febrile patients in Vientiane, Lao PDR. The final method showed promising sensitivities of 66% (95% confidence interval [CI]: 55-76) and 59% (95% CI: 49-68) compared with direct or direct and indirect testing combined, as the respective reference standards (specificities > 95%). Despite these promising diagnostic parameters, a subsequent prospective evaluation in a Lao hospital population (N= 352) showed that the sensitivity was very low (∼30%) compared with qPCR on venous blood samples. The disappointingly low sensitivity does suggest that venous blood samples are preferable for the clinical microbiology laboratory, although BCF might be an alternative if leptospirosis is only suspected postadmission after antibiotics have been used., (© The American Society of Tropical Medicine and Hygiene.)

Leptospirosis is an important zoonotic disease in the Pacific Islands. In Fiji, two successive cyclones and severe flooding in 2012 resulted in outbreaks with 576 reported cases and 7% case-fatality. We conducted a cross-sectional seroprevalence study and used an eco-epidemiological approach to characterize risk factors and drivers for human leptospirosis infection in Fiji, and aimed to provide an evidence base for improving the effectiveness of public health mitigation and intervention strategies. Antibodies indicative of previous or recent infection were found in 19.4% of 2152 participants (81 communities on the 3 main islands). Questionnaires and geographic information systems data were used to assess variables related to demographics, individual behaviour, contact with animals, socioeconomics, living conditions, land use, and the natural environment. On multivariable logistic regression analysis, variables associated with the presence of Leptospira antibodies included male gender (OR 1.55), iTaukei ethnicity (OR 3.51), living in villages (OR 1.64), lack of treated water at home (OR 1.52), working outdoors (1.64), living in rural areas (OR 1.43), high poverty rate (OR 1.74), living <100m from a major river (OR 1.41), pigs in the community (OR 1.54), high cattle density in the district (OR 1.04 per head/sqkm), and high maximum rainfall in the wettest month (OR 1.003 per mm). Risk factors and drivers for human leptospirosis infection in Fiji are complex and multifactorial, with environmental factors playing crucial roles. With global climate change, severe weather events and flooding are expected to intensify in the South Pacific. Population growth could also lead to more intensive livestock farming; and urbanization in developing countries is often associated with urban and peri-urban slums where diseases of poverty proliferate. Climate change, flooding, population growth, urbanization, poverty and agricultural intensification are important drivers of zoonotic disease transmission; these factors may independently, or potentially synergistically, lead to enhanced leptospirosis transmission in Fiji and other similar settings.

Leptospirosis causes significant economic loss within the cattle industry worldwide. Current diagnostic methods are generally inadequate for dealing with large numbers of samples, are outdated, and provide little useful diagnostic and epidemiological information. This aim of this study was to apply a microsphere immunoassay (MIA), utilising Luminex xMap technology, to 200 bovine serum samples to determine this method's usefulness in leptospirosis diagnosis in comparison with the current gold standard, the microscopic agglutination test (MAT). Although MAT is the most widely used laboratory test for the diagnosis of leptospirosis, its reliance on live cultures, subjective interpretation of results and an inability to differentiate between antibody classes, suggest MAT is no longer the best method for the diagnosis of leptospirosis. The results presented in this paper show that MIA was able to determine reactive from non-reactive samples when compared with MAT, and was able to differentiate IgG and IgM classes of antibody. The results suggest increased sensitivity in MIA and the ability to multiplex up to 500 antigens at one time allows for significant improvements in cost-effectiveness as well as a reduced dependency on live cultures. The relatively low cost, high throughput platform and differentiation of antibody class, as shown in previous research, make this assay worthy of consideration for the diagnosis of leptospirosis in small-scale or large-scale bovine populations.

The etiology of fever in rural Lao People's Democratic Republic (Laos) has remained obscure until recently owing to the lack of laboratory facilities. We conducted a study to determine the causes of fever among 229 patients without malaria in Savannakhet Province, southern Laos; 52% had evidence of at least one diagnosis (45% with single and 7% with apparent multiple infections). Among patients with only one diagnosis, dengue (30.1%) was the most common, followed by leptospirosis (7.0%), Japanese encephalitis virus infection (3.5%), scrub typhus (2.6%), spotted fever group infection (0.9%), unspecified flavivirus infection (0.9%), and murine typhus (0.4%). We discuss the empirical treatment of fever in relation to these findings., (© The American Society of Tropical Medicine and Hygiene.)

A microsphere immunoassay (MIA) utilising Luminex xMap technology that is capable of determining leptospirosis IgG and IgM independently was developed. The MIA was validated using 200 human samples submitted for routine leptospirosis serology testing. The traditional microscopic agglutination (MAT) method (now 100 years old) suffers from a significant range of technical problems including a dependence on antisera which is difficult to source and produce, false positive reactions due to auto-agglutination and an inability to differentiate between IgG and IgM antibodies. A comparative validation method of the MIA against the MAT was performed and used to determine the ability of the MIA to detect leptospiral antibodies when compared with the MAT. The assay was able to determine samples in the reactive, equivocal and non-reactive ranges when compared to the MAT and was able to differentiate leptospiral IgG antibodies from leptospiral IgM antibodies. The MIA is more sensitive than the MAT and in true infections was able to detect low levels of antibody in the later stages of the acute phase as well as detect higher levels of IgM antibody earlier in the immune phase of the infection. The relatively low cost, high throughput platform and significantly reduced dependency on large volumes of rabbit antisera make this assay worthy of consideration for any microbiological assay that currently uses agglutination assays.

Background: Scrub typhus (caused by Orientia tsutsugamushi), murine typhus (caused by Rickettsia typhi), and leptospirosis are common causes of febrile illness in Asia; meningitis and meningoencephalitis are severe complications. However, scarce data exist for the burden of these pathogens in patients with CNS disease in endemic countries. Laos is representative of vast economically poor rural areas in Asia with little medical information to guide public health policy. We assessed whether these pathogens are important causes of CNS infections in Laos., Methods: Between Jan 10, 2003, and Nov 25, 2011, we enrolled 1112 consecutive patients of all ages admitted with CNS symptoms or signs requiring a lumbar puncture at Mahosot Hospital, Vientiane, Laos. Microbiological examinations (culture, PCR, and serology) targeted so-called conventional bacterial infections (Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, S suis) and O tsutsugamushi, Rickettsia typhi/Rickettsia spp, and Leptospira spp infections in blood or cerebrospinal fluid (CSF). We analysed and compared causes and clinical and CSF characteristics between patient groups., Findings: 1051 (95%) of 1112 patients who presented had CSF available for analysis, of whom 254 (24%) had a CNS infection attributable to a bacterial or fungal pathogen. 90 (35%) of these 254 infections were caused by O tsutsugamushi, R typhi/Rickettsia spp, or Leptospira spp. These pathogens were significantly more frequent than conventional bacterial infections (90/1051 [9%] vs 42/1051 [4%]; p<0·0001) by use of conservative diagnostic definitions. CNS infections had a high mortality (236/876 [27%]), with 18% (13/71) for R typhi/Rickettsia spp, O tsutsugamushi, and Leptospira spp combined, and 33% (13/39) for conventional bacterial infections (p=0·076)., Interpretation: Our data suggest that R typhi/Rickettsia spp, O tsutsugamushi, and Leptospira spp infections are important causes of CNS infections in Laos. Antibiotics, such as tetracyclines, needed for the treatment of murine typhus and scrub typhus, are not routinely advised for empirical treatment of CNS infections. These severely neglected infections represent a potentially large proportion of treatable CNS disease burden across vast endemic areas and need more attention., Funding: Wellcome Trust UK., (Copyright © 2015 Dittrich et al. Open Access article published under the terms of CC BY. Published by .. All rights reserved.)

Leptospirosis outbreaks have been associated with many common water events including water consumption, water sports, environmental disasters, and occupational exposure. The ability of leptospires to survive in moist environments makes them a high-risk agent for infection following contact with any contaminated water source. Water treatment processes reduce the likelihood of leptospirosis or other microbial agents causing infection provided that they do not malfunction and the distribution networks are maintained. Notably, there are many differences in water treatment systems around the world, particularly between developing and developed countries. Detection of leptospirosis in water samples is uncommonly performed by molecular methods.

The following research reports the emergence of Leptospira borgpetersenii serovar Arborea as the dominant infecting serovar following the summer of disasters and the ensuing clean up in Queensland, Australia during 2011. For the 12 month period (1 January to 31 December) L. borgpetersenii serovar Arborea accounted for over 49% of infections. In response to a flooding event public health officials need to issue community wide announcements warning the population about the dangers of leptospirosis and other water borne diseases. Communication with physicians working in the affected community should also be increased to update physicians with information such as clinical presentation of leptospirosis and other waterborne diseases. These recommendations will furnish public health officials with considerations for disease management when dealing with future disaster management programs.

In leptospirosis patients, haematological abnormalities have been reported. The aim of this study was to determine if neutrophil counts were different between patients known to be infected with a range of leptospiral serovars. The study retrospectively compared the neutrophil counts from the first blood samples taken from 210 leptospirosis patients at first presentation to a Queensland Health hospital. Significant differences (p <0.001) were observed in neutrophil counts across the 11 different infecting serovars. These findings suggest that neutrophil counts may be useful in the development of an algorithm determining the infecting serovar in suspected leptospirosis patients. Further studies are required to delineate host cytokine responses which may suggest the underlying aetiology of the observed differences in neutrophil counts. Such studies would also provide valuable therapeutic insights into treating the disease.

Background: Because of reductions in the incidence of Plasmodium falciparum malaria in Laos, identification of the causes of fever in people without malaria, and discussion of the best empirical treatment options, are urgently needed. We aimed to identify the causes of non-malarial acute fever in patients in rural Laos., Methods: For this prospective study, we recruited 1938 febrile patients, between May, 2008, and December, 2010, at Luang Namtha provincial hospital in northwest Laos (n=1390), and between September, 2008, and December, 2010, at Salavan provincial hospital in southern Laos (n=548). Eligible participants were aged 5-49 years with fever (≥38°C) lasting 8 days or less and were eligible for malaria testing by national guidelines., Findings: With conservative definitions of cause, we assigned 799 (41%) patients a diagnosis. With exclusion of influenza, the top five diagnoses when only one aetiological agent per patient was identified were dengue (156 [8%] of 1927 patients), scrub typhus (122 [7%] of 1871), Japanese encephalitis virus (112 [6%] of 1924), leptospirosis (109 [6%] of 1934), and bacteraemia (43 [2%] of 1938). 115 (32%) of 358 patients at Luang Namtha hospital tested influenza PCR-positive between June and December, 2010, of which influenza B was the most frequently detected strain (n=121 [87%]). Disease frequency differed significantly between the two sites: Japanese encephalitis virus infection (p=0·04), typhoid (p=0·006), and leptospirosis (p=0·001) were more common at Luang Namtha, whereas dengue and malaria were more common at Salavan (all p<0·0001). With use of evidence from southeast Asia when possible, we estimated that azithromycin, doxycycline, ceftriaxone, and ofloxacin would have had significant efficacy for 258 (13%), 240 (12%), 154 (8%), and 41 (2%) of patients, respectively., Interpretation: Our findings suggest that a wide range of treatable or preventable pathogens are implicated in non-malarial febrile illness in Laos. Empirical treatment with doxycycline for patients with undifferentiated fever and negative rapid diagnostic tests for malaria and dengue could be an appropriate strategy for rural health workers in Laos., Funding: Wellcome Trust, WHO-Western Pacific Region, Foundation for Innovative New Diagnostics, US Centers for Disease Control and Prevention

Throughout December 2010 and January 2011, Queensland experienced widespread flooding due to unusually protracted and heavy rainfalls. In mid-January 2011, four individuals from a small community in Central Queensland were hospitalized with leptospirosis. A further five cases were subsequently identified from around Central Queensland, bringing the total to nine. Microscopic agglutination testing found that serovar Arborea (Leptospira borgpetersenii serovar Arborea) was presumptively responsible for leptospirosis in seven of nine confirmed cases. Serovars Hardjo and Australis were identified in samples from two remaining cases. All cases had exposure to flood water. No single exposure source was identified. This is the first reported outbreak of leptospirosis in Central Queensland and the first report of leptospirosis cases associated with flood water inundation in Queensland. Public health authorities should continue to promote awareness of leptospirosis in flood-affected populations. Healthcare providers must maintain a high level of suspicion for leptospirosis during and after flood events.

Introduction: The aim of the study was to compare haemoglobin and red cell counts between patients known to be infected with a range of leptospiral serovars., Methods: The study retrospectively compared the haemoglobin and red cell count results from the first blood samples taken from 207 patients at presentation to a Queensland Health hospital., Results: Significant differences were observed in haemoglobin and red cell counts in those infected with Leptospira interrogans serovars Szwajizak and Canicola when compared with most of the other serovars., Conclusions: These findings suggest that haemoglobin and red cell counts may be useful in differentiating leptospiral serovars in leptospirosis patients.

When data are collected via anonymous Internet surveys, particularly under conditions of obligatory participation (such as with student samples), data quality can be a concern. However, little guidance exists in the published literature regarding techniques for detecting careless responses. Previously several potential approaches have been suggested for identifying careless respondents via indices computed from the data, yet almost no prior work has examined the relationships among these indicators or the types of data patterns identified by each. In 2 studies, we examined several methods for identifying careless responses, including (a) special items designed to detect careless response, (b) response consistency indices formed from responses to typical survey items, (c) multivariate outlier analysis, (d) response time, and (e) self-reported diligence. Results indicated that there are two distinct patterns of careless response (random and nonrandom) and that different indices are needed to identify these different response patterns. We also found that approximately 10%-12% of undergraduates completing a lengthy survey for course credit were identified as careless responders. In Study 2, we simulated data with known random response patterns to determine the efficacy of several indicators of careless response. We found that the nature of the data strongly influenced the efficacy of the indices to identify careless responses. Recommendations include using identified rather than anonymous responses, incorporating instructed response items before data collection, as well as computing consistency indices and multivariate outlier analysis to ensure high-quality data.

Leptospirosis has recently been reported as an emerging disease worldwide, and a seroprevalence study was undertaken in American Samoa to better understand the drivers of transmission. Antibodies indicative of previous exposure to leptospirosis were found in 15.5% of 807 participants, predominantly against three serovars that were not previously known to occur in American Samoa. Questionnaires and geographic information systems data were used to assess behavioral factors and environmental determinants of disease transmission, and logistic regression was used to identify factors associated with infection. Many statistically significant factors were consistent with previous studies, but we also showed a significant association with living at lower altitudes (odds ratio [OR] = 1.53, 95% confidence interval [CI]: 1.03-2.28), and having higher numbers of piggeries around the home (OR = 2.63, 95% CI: 1.52-4.40). Our findings support a multifaceted approach to combating the emergence of leptospirosis, including modification of individual behavior, but importantly also managing the evolving environmental drivers of risk.

Background: Leptospirosis has recently been discussed as an emerging infectious disease in many contexts, including changes in environmental drivers of disease transmission and the emergence of serovars. In this paper, we report the epidemiology of leptospiral serovars from our study of human leptospirosis in American Samoa in 2010, present evidence of recent serovar emergence, and discuss the potential epidemiological and ecological implications of our findings., Methods: Serovar epidemiology from our leptospirosis seroprevalence study in 2010 was compared to findings from a study in 2004. The variation in geographic distribution of the three most common serovars was explored by mapping sero-positive participants to their place of residence using geographic information systems. The relationship between serovar distribution and ecological zones was examined using geo-referenced data on vegetation type and population distribution., Results: Human leptospirosis seroprevalence in American Samoa was 15.5% in 2010, with serological evidence that infection was caused by three predominant serovars (Hebdomadis, LT 751, and LT 1163). These serovars differed from those identified in an earlier study in 2004, and were not previously known to occur in American Samoa. In 2010, serovars also differed in geographic distribution, with variations in seroprevalence between islands and different ecological zones within the main island., Conclusions: Our findings might indicate artefactual emergence (where serovars were long established but previously undetected), but we believe the evidence is more in favour of true emergence (a result of ecological change). Possibilities include changes in interactions between humans and the environment; introduction of serovars through transport of animals; evolution in distribution and/or abundance of animal reservoirs; and environmental changes that favour transmission of particular serovars.Future research should explore the impact of ecological change on leptospirosis transmission dynamics and serovar emergence, and investigate how such new knowledge might better target environmental monitoring for disease control at a public health level.

Global leptospirosis disease burden estimates are hampered by the lack of scientifically sound data from countries with probable high endemicity and limited diagnostic capacities. We describe the seroepidemiologic and clinical characteristics of the leptospirosis outbreak in 2008 in Sri Lanka. Definitive/presumptive case definitions proposed by the World Health Organization Leptospirosis Epidemiology Reference Group were used for case confirmation. Of the 404 possible cases, 155 were confirmed to have leptospirosis. Highest titers of patient seum samples reacted with serovars Pyrogenes (28.7%), Hardjo (18.8%), Javanica (11.5%), and Hebdomadis (11.5%). Sequencing of the 16S ribosomal DNA gene identified six infections: five with Leptospira interrogans and one with L. weilli. In this patient population, acute renal failure was the main complication (14.8%), followed by myocarditis (7.1%) and heart failure (3.9%). The case-fatality rate was 1.3%. This report strengthens the urgent need for increasing laboratory diagnostic capabilities to determine the causes of epidemic and endemic infectious diseases in Sri Lanka, a finding relevant to other tropical regions.

Identification of wild animals that harbour the causative leptospires, and the identification of the most important of these 'wild reservoirs' (in terms of threat to human health), are key factors in the epidemiology of human leptospirosis. In an epidemiological investigation in the Australian state of Queensland, in 2007-2008, samples were collected from fruit bats (Pteropus conspicillatus) and rodents (to investigate the potential role of fruit bats in the maintenance and transmission of leptospires to ground-dwelling rodents) and checked for pathogenic leptospires. The results of these studies have now been carefully analysed in attempts to see which method of detection and type of test sample were best. The effects of pentobarbitone sodium used to euthanize wild mammals before collection of necropsy samples, on the survival and detection of leptospires in vitro, were also explored. In the earlier field investigation, serum, renal tissue and urine were collected from wild mammals, for the detection of pathogenic leptospires by culture, the microscopic agglutination test (MAT), real-time PCR and silver impregnation of smears. Although 27.6% of the rodents investigated were found leptospire-positive, culture only yielded four isolates, probably because many cultures were contaminated. The main aims of the present study were to quantify the performance of the individual diagnostic tests and examine the reasons behind the high incidence of culture contamination. The results of sensitivity and specificity analyses for the different diagnostic tests indicated that isolation by culture (the definitive diagnostic test for leptospiral shedding) had perfect (100%) sensitivity when compared with the results of the PCR but a low specificity (40%). The MAT performed poorly, with a sensitivity of 50% when compared against the results of culture. The prevalence of leptospiral carriage revealed by the PCR-based investigation of kidney and urine samples (59.2%) was higher than that revealed using any other method and far higher than the 2.0% revealed by culture. The results of the culture of renal tissue agreed fairly well with those of the PCR-based investigation of such tissue, with a Cohen's unweighted kappa coefficient (κ) of 0.5 (P = 0.04). The levels of agreement between other pairs of tests were generally poor. The presence of pentobarbitone sodium, at final concentrations of 27.8 or 167 mg/ml, did not affect the viability or the detection of leptospires in culture, and is therefore unlikely to reduce the chances of isolating leptospires from an animal that has been euthanized with the compound. It appears that collecting multiple samples from each mammal being checked will improve the chances of detecting leptospires (and reduce the chances of reporting an inconclusive result for any of the mammals). For the identification of a leptospiral carrier, however, the use of just two detection methods (culture and PCR) and one type of sample (renal tissue) may give adequate sensitivity and specificity. Given the robustness of PCR to contamination and its high sensitivity (it can give a positive result when DNA from just two leptospiral cells is present in the sample), a PCR-based serotyping method, to allow the combined detection and characterisation of leptospires from field isolates, would be extremely useful.

Ticks are obligate haematophagous ectoparasites of various animals, including humans, and are abundant in temperate and tropical zones around the world. They are the most important vectors for the pathogens causing disease in livestock and second only to mosquitoes as vectors of pathogens causing human disease. Ticks are formidable arachnids, capable of not only transmitting the pathogens involved in some infectious diseases but also of inducing allergies and causing toxicoses and paralysis, with possible fatal outcomes for the host. This review focuses on tick paralysis, the role of the Australian paralysis tick Ixodes holocyclus, and the role of toxin molecules from this species in causing paralysis in the host.

Hendra virus (HeV) was first isolated in 1994, from a disease outbreak involving at least 21 horses and two humans in the Brisbane suburb of Hendra, Australia. The affected horses and humans all developed a severe but unidentified respiratory disease that resulted in the deaths of one of the human cases and the deaths or putting down of 14 of the horses. The virus, isolated by culture from a horse and the kidney of the fatal human case, was initially characterised as a new member of the genus Morbillivirus in the family Paramyxoviridae. Comparative sequence analysis of part of the matrix protein gene of the virus and the discovery that the virus had an exceptionally large genome subsequently led to HeV being assigned to a new genus, Henipavirus, along with Nipah virus (a newly emergent virus in pigs). The regular outbreaks of HeV-related disease that have occurred in Australia since 1994 have all been characterised by acute respiratory and neurological manifestations, with high levels of morbidity and mortality in the affected horses and humans. The modes of transmission of HeV remain largely unknown. Although fruit bats have been identified as natural hosts of the virus, direct bat-horse, bat-human or human-human transmission has not been reported. Human infection can occur via exposure to infectious urine, saliva or nasopharyngeal fluid from horses. The treatment options and efficacy are very limited and no vaccine exists. Reports on the outbreaks of HeV in Australia are collated in this review and the available data on the biology, transmission and detection of the pathogen are summarized and discussed.

Although antileptospiral antibodies and leptospiral DNA have been detected in Australian fruit bats, the role of such bats as infectious hosts for the leptospires found in rodents and humans remains unconfirmed. A cohort-design, replicated survey was recently conducted in Far North Queensland, Australia, to determine if the abundance and leptospiral status of rodents were affected by association with colonies of fruit bats (Pteropus conspicillatus spp.) via rodent contact with potentially infectious fruit-bat urine. In each of four study areas, a 'colony site' that included a fruit-bat colony and the land within 1500 m of the colony was compared with a 'control site' that held no fruit-bat colonies and was >2000 m from the nearest edge of the colony site. Rodents were surveyed, for a total of 2400 trap-nights, over six sampling sessions between September 2007 and September 2008. A low abundance of rodents but a high carriage of leptospires in the rodents present were found to be associated with proximity to a fruit-bat colony. For example, means of 0·4 and 2·3 fawn-footed melomys (Melomys cervinipes) were collected/100 trap-nights at sites with and without fruit-bat colonies, respectively (P<0·001), but the corresponding prevalences of leptospiral carriage were 100% and 3·6% (P<0·001). Such trends were consistent across all of the sampling sessions but not across all of the sampling sites. Leptospires were not isolated from fruit bats by culture, and the role of such bats in the transmission of leptospires to rodents cannot be confirmed. The data collected do, however, indicate the existence of a potential pathway for transmission of leptospires from fruit bats to rodents, via rodent contact with infectious fruit-bat urine. Fruit bats may possibly be involved in the ecology of leptospires (including emergent serovars), as disseminators of pathogens to rodent populations. Stringent quantitative risk analysis of the present and similar data, to explore their implications in terms of disease prevalence and wildlife population dynamics, is recommended.

Since its discovery in a juvenile black flying fox (Pteropus alecto) in 1996, Australian bat lyssavirus (ABLV) has become the cause of a potentially important emerging disease for health authorities in Australia, with two human deaths (one in 1996 and one in 1998) attributed to the virus in the north-eastern state of Queensland. In Australia, the virus has been isolated from all four species of flying fox found on the mainland (i.e. P. alecto, P. scapulatus, P. poliocephalus and P. conspicillatus) as well as a single species of insectivorous bat (Saccolaimus flaviventris). Australian bat lyssavirus belongs to the Lyssavirus genus and is closely related, genetically, to the type strain of Rabies virus (RABV). Clinically, patients infected with ABLV have displayed the 'classical' symptoms of rabies and a similar disease course. This similarity has led to the belief that the infection and dissemination of ABLV in the body follows the same pathways as those followed by RABV. Following the two ABLV-related deaths in Queensland, protocols based on the World Health Organization's guidelines for RABV prophylaxis were implemented and, presumably in consequence, no human infection with ABLV has been recorded since 1998. ABLV will, however, probably always have an important part to play in the health of Australians as the density of the human population in Australia and, consequently, the level of interaction between humans and flying foxes increase.

Mosquito-borne diseases continue to be a serious public-health concern in Australia. Endemic alphaviruses (including Ross River and Barmah Forest viruses) account for the majority of the arboviral notifications, while some flaviviruses (Murray Valley encephalitis, Japanese encephalitis and Kunjin viruses) cause occasional outbreaks of encephalitis. Dengue epidemics are increasing in frequency in northern Queensland, with the largest outbreak in 50 years occurring during the 2008-2009 wet season. Of great concern are the threats posed by the importation of exotic arboviruses, such as West Nile, chikungunya and Rift Valley fever viruses, the introduction of exotic vectors, and the potential range expansion of key Australian vectors. Environmental and anthropogenic influences provide additional uncertainty regarding the future impact of mosquito-borne pathogens in Australia. This review discusses the trends, threats and challenges that face the management of mosquito-borne disease in Australia. Topical mosquito-borne pathogens of biosecurity and public-health concern, and the potential impacts of environmental and global trends, are discussed. Finally, a short overview of the public-health response capability in Australia is provided.

Leptospira borgpetersenii serovar Arborea is an emerging cause of leptospirosis in Australia. It was not previously recognized as an endemic serovar before the 1990s, but at that point, human infections with the serovar increased significantly. Using fluorescent-amplified fragment-length polymorphism (FAFLP) molecular typing, human and rodent isolates were compared genetically. Typing revealed 11 unique profiles among the 23 isolates examined; however, there was no clonality revealed between the human and rodent isolates. There was clonality among rodent isolates from geographically related areas. This study highlights the utility of Leptospira culture combined with FAFLP for the examination of the epidemiology of this disease.

Human leptospirosis is a zoonotic disease of global importance that causes significant morbidity and mortality, particularly in developing nations. In this review, the history, epidemiology, transmission, clinical presentation and treatment of this disease, and its impact in Australia, are discussed. Central to this review is the delineation of diagnostic methods for the disease and the challenges that this disease presents for both the clinician and diagnostic laboratory. This information should furnish clinicians with an updated tool to help overcome a number of problems associated with the diagnosis of leptospirosis.

Flooding and heavy rainfall have been associated with numerous outbreaks of leptospirosis around the world. With global climate change, extreme weather events such as cyclones and floods are expected to occur with increasing frequency and greater intensity and may potentially result in an upsurge in the disease incidence as well as the magnitude of leptospirosis outbreaks. In this paper, we examine mechanisms by which climate change can affect various ecological factors that are likely to drive an increase in the overall incidence as well as the frequency of outbreaks of leptospirosis. We will discuss the geographical areas that are most likely to be at risk of an increase in leptospirosis disease burden owing to the coexistence of climate change hazard risk, environmental drivers of leptospirosis outbreaks, local socioeconomic circumstances, and social and demographic trends. To reduce this disease burden, enhanced surveillance and further research is required to understand the environmental drivers of infection, to build capacity in emergency response and to promote community adaptation to a changing climate., (Copyright © 2010 Royal Society of Tropical Medicine and Hygiene.)

Ciguatera poisoning is a food-borne neuro-intoxication caused by consumption of finfish that have accumulated ciguatoxins in their tissues. Ciguatera is a distressing and sometimes disabling condition that presents with a self-limiting though occasionally severe gastro-intestinal illness, progressing to a suite of aberrant sensory symptoms. Recovery can take from days to years; second and subsequent attacks may manifest in a more severe illness. Ciguatera remains largely a pan-tropical disease, although tourism and export fish markets facilitate increased presentation in temperate latitudes. While ciguatera poisoning in the South Pacific was recognised and eloquently described by seafarers in the 18th Century, it remains a public-health challenge in the 21st Century because there is neither a confirmatory diagnostic test nor a reliable, low-cost screening method to ascertain the safety of suspect fish prior to consumption. A specific antidote is not available, so treatment is largely supportive. The most promising pharmacotherapy of recent decades, intravenous mannitol, has experienced a relative decline in acceptance after a randomized, double-blind trial failed to confirm its efficacy. Some questions remain unanswered, however, and the use of mannitol for the treatment of acute ciguatera poisoning arguably deserves revisiting. The immunotoxicology of ciguatera is poorly understood, and some aspects of the epidemiology and symptomatology of ciguatera warrant further enquiry.

High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) is a novel technology that has emerged as a possible method to characterise leptospires to serovar level. RAPD-HRM has recently been used to measure intra-serovar convergence between strains of the same serovar as well as inter-serovar divergence between strains of different serovars. The results indicate that intra-serovar heterogeneity and inter-serovar homogeneity may limit the application of RAPD-HRM in routine diagnostics. They also indicate that genetic attenuation of aged, high-passage-number isolates could undermine the use of RAPD-HRM or any other molecular technology. Such genetic attenuation may account for a general decrease seen in titres of rabbit hyperimmune antibodies over time. Before RAPD-HRM can be further advanced as a routine diagnostic tool, strains more representative of the wild-type serovars of a given region need to be identified. Further, RAPD-HRM analysis of reference strains indicates that the routine renewal of reference collections, with new isolates, may be needed to maintain the genetic integrity of the collections.

A new test for pathogenic Leptospira isolates, based on RAPD-PCR and high-resolution melt (HRM) analysis (which measures the melting temperature of amplicons in real time, using a fluorescent DNA-binding dye), has recently been developed. A characteristic profile of the amplicons can be used to define serovars or detect genotypes. Ten serovars, of leptospires from the species Leptospira interrogans (serovars Australis, Robinsoni, Hardjo, Pomona, Zanoni, Copenhageni and Szwajizak), L. borgpetersenii (serovar Arborea), L. kirschneri (serovar Cynopteri) and L. weilii (serovar Celledoni), were typed against 13 previously published RAPD primers, using a real-time cycler (the Corbett Life Science RotorGene 6000) and the optimised reagents from a commercial kit (Quantace SensiMix). RAPD-HRM at specific temperatures generated defining amplicon melt profiles for each of the tested serovars. These profiles were evaluated as difference-curve graphs generated using the RotorGene software package, with a cut-off of at least 8 'U' (plus or minus). The results demonstrated that RAPD-HRM can be used to measure serovar diversity and establish identity, with a high degree of stability. The characterisation of Leptospira serotypes using a DNA-based methodology is now possible. As an objective and relatively inexpensive and rapid method of serovar identification, at least for cultured isolates, RAPD-HRM assays show convincing potential.

Magnesium imbalance in leptospirosis has, for the most part, been neglected by the medical and leptospirosis communities. In a recent, retrospective study, serum concentrations of magnesium were followed in 15 patients with severe leptospirosis. The results revealed that 14 of the 15 patients developed hypomagnesaemia at some time during the first 10 days of their illness. In severely ill patients, such magnesium deficiency can worsen clinical outcome. Magnesium concentrations may affect a number of organ systems and mental status. Since altered mental status in leptospirosis is a poor prognostic indicator, it is suggested that serum concentrations of magnesium be monitored closely in patients with leptospirosis. Any hypomagnesaemia can then be treated promptly, in an effort to reduce the morbidity and mortality attributable to the disease.

To identify the hosts of mosquitoes collected from urban and peri-urban habitats in eastern Australia, 1,180 blood fed mosquitoes representing 15 species were analyzed by enzyme-linked immunosorbent assay and molecular techniques. Four common and epidemiologically important species could be classified according to their host-feeding patterns: Aedes aegypti was anthropophilic, Ae. vigilax was mammalophilic, Culex quinquefasciatus was ornithophilic, and Cx. annulirostris was opportunistic, readily feeding on birds and mammals. Mitochondrial cytochrome b DNA sequence data showed that more than 75% of avian blood meals identified from Cx. annulirostris collected from Brisbane, Newcastle, and Sydney originated from ducks (Order Anseriformes, Family Anatidae). More than 75% of avian blood meals from Cx. quinquefasciatus from Cairns belonged to one of three Passerine species, namely Sphecotheres vieilloti (figbird), Sturnus tristis (common myna), and Philemon buceroides (helmeted friarbird). This study demonstrates associations between vectors in Australia and vertebrate hosts of endemic and exotic arboviruses.

Leptospiral pathogens have a world-wide distribution and cause a spectrum of disease ranging from a mild, influenza-like illness to Weil's disease, which manifests itself in multi-organ failure. Recently, Leptospira-reactive sera from 40 leptospirosis patients were investigated in an ELISA designed to detect antibodies to the human glomerular basement membrane (GBM). The aim was to determine if host-derived leptospiral immunoglobulins cross-react with proteins in the human GBM, so facilitating the development of Goodpasture's syndrome. As all 40 sera were found negative in the anti-GBM ELISA, the hypothesis that, during the immune phase of leptospirosis, patients are at risk of developing Goodpasture's syndrome was not supported. Further work is required to determine if leptospirosis is a risk factor in the development of any other pulmonary-renal syndromes that are associated with auto-immune diseases, such as Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, Behçet's disease, IgA nephropathy and systemic lupus erythematosus.

Lymphocyte counts in patients with leptospirosis have been shown to be variable. This study retrospectively compared lymphocyte counts from the first blood samples taken following hospital presentation in patients with leptospirosis who were either (i) IgM non-reactive, (ii) IgM reactive and microscopic agglutination test (MAT) non-reactive or (iii) IgM and MAT reactive in an effort to determine whether differences in lymphocyte counts are observed in the acute and immune phase of leptospirosis. Statistical differences in lymphocyte counts were observed between the three groups. In conclusion, this study has shown that the phase of leptospiral infection may affect patient lymphocyte counts.