The emergence of Leptospira borgpetersenii serovar Arborea in Queensland, Australia, 2001 to 2013

BACKGROUND: Leptospirosis is an emerging infectious disease, with increasing frequency and severity of outbreaks, changing epidemiology of populations at risk, and the emergence of new serovars. Environmental drivers of disease transmission include flooding, urbanisation, poor sanitation, changes in land use and agricultural practices, and socioeconomic factors. In Queensland, human infection with Leptosira borgpetersenii serovar Arborea was first reported in 2001. This study aims to report the emergence of serovar Arborea in Queensland from 2001 to 2013, and investigate potential risk factors for infection and drivers of emergence. METHODS: Data on laboratory-confirmed cases of human leptospirosis in Queensland were obtained from the enhanced surveillance system at the WHO/FAO/OIE Collaborating Centre for Reference and Research on Leptospirosis in Brisbane, Australia. The changing epidemiology of serovar Arborea from 2001 to 2003 was described with respect to case numbers, proportion of leptospirosis cases attributed to the serovar, and geographic distribution. Differences in risk factors for the most common serovars were compared. RESULTS: During this period, 1289 cases of leptospirosis were reported, including 233 cases attributed to serovar Arborea. Risk factors for infection include male gender (91 % of cases), occupation, and recreational exposure. Most common occupations recorded were banana workers (28.4 %), meat workers (7.2 %), dairy farmers (5.8 %), graziers/stockmen (5.5 %), 'other agricultural/rural workers' (16.4 %), and tourists or tourism operators (4.6 %). Time trend analysis showed that while non-Arborea cases decreased over the study period, Arborea cases increased by 3.4 cases per year. The proportion of annual cases attributed to Arborea peaked at 49 % in 2011 after unprecedented flooding in Queensland. Mapping of cases by residential location showed expansion of the geographic range of serovar Arborea, concentrating mostly around Brisbane, Cairns and Innisfail. Serovars varied significantly between ages and occupational groups, and serovar Arborea was most strongly associated with 'other agricultural/rural workers'. CONCLUSIONS: Leptospira borgpetersenii serovar Arborea has been emerging in Queensland since 2001, with increase in case numbers, the proportion of leptospirosis infections attributed to the serovar, as well as expansion of its geographic distribution. Reasons for this emergence are unknown, but climatic factors and environmental change are likely to have played important roles

Review of leptospirosis notifications in Queensland and Australia: January 1998 - June 1999

The World Health Organization/Food and Agricultural Organization Collaborating Centre for Reference and Research on Leptospirosis, Western Pacific Region, accredited since 1958, is part of Queensland Health Scientific Services, which provide tertiary level support in epidemiology, surveillance, training and diagnosis for hospitals and pathology laboratories across the State. Databases for leptospirosis on a global, Australian and State-wide basis are maintained on site and support public health authorities in Australia, WHO and the International Leptospirosis Society. Queensland data collated and analysed from leptospirosis questionnaires, and a brief overview of Australian data based on questionnaire responses for notified cases from 1998 to June 1999, are summarised. The increase in leptospirosis notifications (77%) during 1998 possibly signalled greater awareness of the disease by clinicians. There was a significant increase in leptospirosis notifications for children and students and a high rate of hospitalisation of cases. An outbreak in North Queensland during the first half of 1999 resulted in 184 notifications with over 50% of cases hospitalised. Polymorphic presentation of the disease with severe pulmonary haemorrhage is associated in particular with the serovar australis. Serovar zanoni continues to be a major cause of severe clinical leptospirosis. Several cases were diagnosed in tourists. One of these cases presented with severe respiratory distress and required 14 days in hospital

Development of a Multiple-Locus Variable number of tandem repeat Analysis (MLVA) for Leptospira interrogans and its application to Leptospira interrogans serovar Australis isolates from Far North Queensland, Australia

BACKGROUND: Leptospirosis is a zoonotic disease caused by the genus, Leptospira. Leptospira interrogans is the most common genomospecies implicated in the disease. Epidemiological investigations are needed to distinguish outbreak situations or to trace reservoirs of the organisms. Current methodologies used for typing Leptospira have significant drawbacks. The development of an easy to perform yet high resolution method is needed for this organism. METHODS: In this study we have searched the available genomic sequence of L. interrogans serovar Copenhageni strain Fiocruz L1-130 for the presence of tandem repeats [1]. These repeats were evaluated against reference strains for diversity. Six loci were selected to create a Multiple Locus Variable Number of Tandem Repeats (VNTR) Analysis (MLVA) to explore the genetic diversity within L. interrogans serovar Australis clinical isolates from Far North Queensland. RESULTS: The 39 reference strains used for the development of the method displayed 39 distinct patterns. Diversity Indexes for the loci varied between 0.80 and 0.93 and the number of repeat units at each locus varied between less than one to 52 repeats. When the MLVA was applied to serovar Australis isolates three large clusters were distinguishable, each comprising various hosts including Rattus species, human and canines. CONCLUSION: The MLVA described in this report, was easy to perform, analyse and was reproducible. The loci selected had high diversity allowing discrimination between serovars and also between strains within a serovar. This method provides a starting point on which improvements to the method and comparisons to other techniques can be made

Identification of pathogenic Leptospira species by conventional or real-time PCR and sequencing of the DNA gyrase subunit B encoding gene

BACKGROUND: Leptospira is the causative genus of the disease, leptospirosis. Species identification of pathogenic Leptospira in the past was generally performed by either DNA-DNA hybridisation or 16s rRNA gene sequencing. Both methods have inherent disadvantages such as the need for radio-labelled isotopes or significant homology between species. A conventional and real-time PCR amplification and sequencing method was developed for an alternate gene target: DNA gyrase subunit B (gyrB). Phylogenetic comparisons were undertaken between pathogenic Leptospira 16srRNA and gyrB genes using clustering and minimum evolution analysis. In addition 50 unidentified Leptospira isolates were characterised by gyrB sequencing and compared with conventional 16s rRNA sequencing. RESULTS: A conventional and real-time PCR methodology was developed and optimised for the amplification of the gyrB from pathogenic Leptospira species. Non pathogenic and opportunistic Leptospira species such as L. fainei and L. broomi were not amplified. The gyrB gene shows greater nucleotide divergence (3.5% to 16.1%) than the 16s rRNA gene (0.1% to 1.4%). Minimum evolution analysis reveals that the gyrB has a different evolution topology for L. kirschneri and L. interrogans. When the two genes were compared for the identification of the 50 unknown isolates there was 100% agreement in the results. CONCLUSION: This research has successfully developed a methodology for the identification of pathogenic Leptospira using an alternate gene to 16s rRNA. The gyrB encoding gene shows higher nucleotide/evolutionary divergence allowing for superior identification and also the potential for the development of DNA probe based identification

The role of pre-sleep cognitions in adolescent sleep-onset problems

© 2018 Elsevier BV. This manuscript version is made available under the CC-BY-NC-ND 4.0 license: http://creativecommons.org/licenses/by-nc-nd/4.0/ This author accepted manuscript is made available following 12 month embargo from date of publication (March 2018) in accordance with the publisher’s archiving policy.Study Objectives: To examine the relationship between pre-sleep cognitions and sleep-onset difficulties in an adolescent sample. Methods: Participants were 385 students (59% male) from grades 9 to 11, between 13 and 18 yrs old (M=15.6, SD=1.0), from 8 co-educational high schools of varied socioeconomic status in metropolitan Adelaide, South Australia. The cross-sectional study used a questionnaire battery including the Sleep Anticipatory Anxiety Questionnaire - Adolescent Version [SAAQ-A], completed during school time, followed by 8 days of sleep diary completion and wearing wrist actigraphy to obtain subjective and objective sleep onset latency [SOL]. Results: Significant relationships were found between somatic arousal (SAAQ-A subscale) and objective SOL, and between sleep-related cognitions (SAAQ-A subscale) and subjective SOL and SOL overestimation (sleep misperception). No relationships were found between subjective SOL and somatic or rehearsal and planning cognitions. Objective SOL was not related to rehearsal and planning, or sleep-related cognition scores, and sleep misperception had no relationship with somatic, and rehearsal and planning cognition scores. Conclusions: These findings are similar to those in clinical adult populations, but also notably different, for example the lack of association between negative sleep-related pre-sleep cognitions and objective sleep difficulty. This study’s results provide a basis for existing relationships between negative pre-sleep cognitions and subjective and objective sleep difficulties in this population to be examined causally in more detail

Il nome e il sangue secondo Quinto Smirneo. Riprese e trasformazioni di un motivo del duello eroico

The aim of this essay is to examine the rules of duelling in Greek epic poetry from Homer and Arctinus to Quintus Smyrnaeus, within the perspective of composition by theme, which make it possible to identify heroes' ancestry as a specific motif of the epic duel

Antibodies to Leptospira among blood donors in higher-risk areas of Australia: Possible implications for transfusion safety

BACKGROUND: Leptospirosis is one of the most common bacterial zoonoses worldwide, and clinical manifestations range from asymptomatic infection to acute febrile illness, multi-organ failure and death. Asymptomatic, acute bacteraemia in a blood donor provides a potential for transfusion-transmission, although only a single such case from India has been recorded. Human leptospirosis is uncommon in developed countries; however, the state of Queensland in Australia has one of the highest rates among developed countries, especially after increased rainfall. This study examined the prevalence of antibodies to Leptospira spp. in blood donors residing in higher-risk areas of Australia, to evaluate the appropriateness of current blood safety guidelines. MATERIALS AND METHODS: Plasma samples collected from blood donors residing in higher-risk areas of Australia during 2009 and 2011 were included in the study. All samples were tested for the presence of antibodies to 22 leptospiral serovars using the microscopic agglutination test. RESULT: No sample had antibody titres suggestive of a current or recent infection, however, seven samples (1.44%, 95% CI: 0.38-2.50%) had titres suggestive of a past infection. DISCUSSION: This study provides data that may support the appropriateness of current relevant donor selection policies in Australia. Given that the risk profile for leptospirosis is expanding and that the infection is likely to become more prevalent with climate change, this disease may become more of a concern for transfusion safety in the futur

Causes of non-malarial fever in Laos: a prospective study

Background Because of reductions in the incidence of Plasmodium falciparum malaria in Laos, identifi cation 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 defi nitions of cause, we assigned 799 (41%) patients a diagnosis. With exclusion of infl uenza, the top fi ve diagnoses when only one aetiological agent per patient was identifi ed 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 infl uenza PCR-positive between June and December, 2010, of which infl uenza B was the most frequently detected strain (n=121 [87%]). Disease frequency diff ered signifi cantly 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 ofl oxacin would have had signifi cant effi cacy for 258 (13%), 240 (12%), 154 (8%), and 41 (2%) of patients, respectively. Interpretation Our fi ndings suggest that a wide range of treatable or preventable pathogens are implicated in nonmalarial febrile illness in Laos. Empirical treatment with doxycycline for patients with undiff erentiated 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 Pacifi c Region, Foundation for Innovative New Diagnostics, US Centers for Disease Control and Prevention

A quantitative PCR (TaqMan) assay for pathogenic Leptospira spp

BACKGROUND: Leptospirosis is an emerging infectious disease. The differential diagnosis of leptospirosis is difficult due to the varied and often "flu like" symptoms which may result in a missed or delayed diagnosis. There are over 230 known serovars in the genus Leptospira. Confirmatory serological diagnosis of leptospirosis is usually made using the microscopic agglutination test (MAT) which relies on the use of live cultures as the source of antigen, often performed using a panel of antigens representative of local serovars. Other techniques, such as the enzyme linked immunosorbent assay (ELISA) and slide agglutination test (SAT), can detect different classes of antibody but may be subject to false positive reactions and require confirmation of these results by the MAT. METHODS: The polymerase chain reaction (PCR) has been used to detect a large number of microorganisms, including those of clinical significance. The sensitivity of PCR often precludes the need for isolation and culture, thus making it ideal for the rapid detection of organisms involved in acute infections. We employed real-time (quantitative) PCR using TaqMan chemistry to detect leptospires in clinical and environmental samples. RESULTS AND CONCLUSIONS: The PCR assay can be applied to either blood or urine samples and does not rely on the isolation and culture of the organism. Capability exists for automation and high throughput testing in a clinical laboratory. It is specific for Leptospira and may discriminate pathogenic and non-pathogenic species. The limit of detection is as low as two cells