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In south-eastern Australia, the pelodryadid Litoria aurea Group (sensu Tyler & Davies 1978) comprises three species: Litoria aurea (Lesson, 1829), Litoria raniformis (Keferstein, 1867), and Litoria castanea (Steindachner, 1867). All three species have been subject to declines over recent decades and taxonomic uncertainty persists among populations on the tablelands in New South Wales. We address the systematics of the Group by analysing mitochondrial and nuclear DNA sequences to assess divergence in the Litoria raniformis from across its current range in New South Wales (NSW), Victoria, South Australia (SA) and Tasmania. We also included samples of Litoria castanea from a recently rediscovered population in the southern tablelands of NSW. Our phylogenetic and population genetic analyses show that Litoria raniformis comprises northern and southern lineages, showing deep mitochondrial DNA sequence divergence (7% net average sequence divergence) and can be diagnosed by fixed allelic differences at more than 4,000 SNP loci. Samples of the northern lineage were collected from the Murray-Darling Basin while those of the southern lineage were collected from south-eastern South Australia, southern and south-eastern Victoria and Tasmania. Analysis of the morphology and bioacoustics did not unequivocally delineate the two lineages. The presence of a hybrid backcross individual in western Victoria at the northern margin of the southern lineage, leads us to assign sub-species status to the two lineages, L. r. raniformis for the northern lineage and L. r. major for the southern lineage. Our data do not unequivocally resolve the taxonomic status of L. castanea which will require molecular genetic analyses of museum vouchers from those parts of the range where L. castanea and L. raniformis are no longer extant. Our data also suggest that human mediated movement of frogs may have occurred over the past 50 years. Our genotyping of vouchers collected in the 1970s from the Mount Lofty Ranges in South Australia detected mitochondrial haplotypes of both sub-species and SNP analysis showed that a single Tasmanian specimen was a backcross with L. r. raniformis ancestry. Movement of L. r. raniformis into Tasmania and both sub-species into the Mount Lofty Ranges are not likely due to passive movements of animals through agricultural commerce, but due to the attractiveness of the species as pets and subsequent escapes or releases, potentially of the larval life stage.

The status of many amphibian populations remains unclear due to undetected declines driven by disease and difficulties in obtaining accurate population estimates. Here, we used genome complexity reduction-based sequencing technology to study the poorly understood Littlejohn’s treefrog, Litoria littlejohni across its fragmented distribution in eastern Australia. We detected five identifiable genetic clusters, with moderate to strong genetic isolation. At a regional scale, population isolation was likely driven by population crashes, resulting in small populations impacted by founder effects. Moderate genetic isolation was detected among populations on the Woronora Plateau despite short distances between population clusters. Evidence of recent declines was apparent in three populations that had very small effective population size, reduced genetic diversity and high inbreeding values. The rates of inbreeding detected in these populations combined with their small size leave these populations at elevated risk of extinction. The Cordeaux Cluster was identified as the most robust population as it was the largest and most genetically diverse. This study exemplifies the value of employing genetic methods to study rare, cryptic species. Despite low recapture rates using traditional capture-recapture demographic methods, we were able to derive population estimates, describe patterns of gene flow, and demonstrate the need for urgent conservation management.

Bovine respiratory disease (BRD) is a major health problem within the global cattle industry. This disease has a complex aetiology, with viruses playing an integral role. In this study, metagenomics was used to sequence viral nucleic acids in the nasal swabs of BRD-affected cattle. The viruses detected included those that are well known for their association with BRD in Australia (bovine viral diarrhoea virus 1), as well as viruses known to be present but not fully characterised (bovine coronavirus) and viruses that have not been reported in BRD-affected cattle in Australia (bovine rhinitis, bovine influenza D, and bovine nidovirus). The nasal swabs from a case–control study were subsequently tested for 10 viruses, and the presence of at least one virus was found to be significantly associated with BRD. Some of the more recently detected viruses had inconsistent associations with BRD. Full genome sequences for bovine coronavirus, a virus increasingly associated with BRD, and bovine nidovirus were completed. Both viruses belong to the Coronaviridae family, which are frequently associated with disease in mammals. This study has provided greater insights into the viral pathogens associated with BRD and highlighted the need for further studies to more precisely elucidate the roles viruses play in BRD.

The status of many amphibian populations remains unclear due to undetected declines driven by disease and difficulties in obtaining accurate population estimates. Here, we used genetic data (Single Nucleotide Polymorphisms) to investigate Australia’s poorly understood Littlejohn’s treefrog, Litoria littlejohni across its fragmented distribution. We detected five identifiable genetic clusters, with moderate to strong genetic isolation. At a regional scale, population isolation likely of population crashes, resulting in small populations with evidence of founder effects. Moderate genetic isolation was detected among populations on the Woronora Plateau despite short distances between population clusters. Evidence of recent declines was apparent in three populations that had very small effective population size, reduced genetic diversity and high inbreeding values. The rates of inbreeding detected in these populations combined with their small size leave these populations at elevated risk of extinction. The Cordeaux Cluster was identified as the most robust population as it was the largest and most genetically diverse. This study exemplifies the value of employing genetic methods to study rare, cryptic species. Despite low recapture rates using traditional capture-recapture demographic methods, we were able to derive population estimates, describe patterns of gene flow, and demonstrate the need for urgent conservation management.

Key Points Multimerization of p67C Ag as nanoparticle increases its immunogenicity. Vaccine efficacy of p67C Ag increases delivered as nanoparticles., Visual Abstract, East Coast fever (ECF), caused by Theileria parva, is the most important tick-borne disease of cattle in sub-Saharan Africa. Practical disadvantages associated with the currently used live-parasite vaccine could be overcome by subunit vaccines. An 80-aa polypeptide derived from the C-terminal portion of p67, a sporozoite surface Ag and target of neutralizing Abs, was the focus of the efforts on subunit vaccines against ECF and subjected to several vaccine trials with very promising results. However, the vaccination regimen was far from optimized, involving three inoculations of 450 μg of soluble p67C (s-p67C) Ag formulated in the Seppic adjuvant Montanide ISA 206 VG. Hence, an improved formulation of this polypeptide Ag is needed. In this study, we report on two nanotechnologies that enhance the bovine immune responses to p67C. Individually, HBcAg-p67C (chimeric hepatitis B core Ag virus-like particles displaying p67C) and silica vesicle (SV)–p67C (s-p67C adsorbed to SV-140-C18, octadecyl-modified SVs) adjuvanted with ISA 206 VG primed strong Ab and T cell responses to p67C in cattle, respectively. Coimmunization of cattle (Bos taurus) with HBcAg-p67C and SV-p67C resulted in stimulation of both high Ab titers and CD4 T cell response to p67C, leading to the highest subunit vaccine efficacy we have achieved to date with the p67C immunogen. These results offer the much-needed research depth on the innovative platforms for developing effective novel protein-based bovine vaccines to further the advancement.

Abortive infection systems (Abi) are phage resistance systems that can be prophage-encoded. Here, two genes encoding an Abi system were identified on a prophage sequence contained by the chromosome of the Levilactobacillus brevis strain UCCLBBS124. This Abi system is similar to the two-component AbiL system encoded by Lactococcus lactis biovar. diacetylactis LD10-1. The UCCLBBS124 prophage-derived Abi system (designated here as AbiL124) was shown to exhibit specific activity against phages infecting L. brevis and L. lactis strains. Expression of the AbiL124 system was shown to cause reduction in the efficiency of plaquing and cell lysis delay for phages of both species.

The MilA ELISA has been identified as a highly effective diagnostic tool for the detection of Mycoplasma bovis specific antibodies and has been validated for serological use in previous studies. This study aimed to estimate the optimal cut-off and corresponding estimates of diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the MilA ELISA for testing bovine serum. Serum samples from 298 feedlot cattle from 14 feedlots across four Australian states were tested on entry into the feedlot and approximately 42 days later. The paired serum samples were tested with the MilA ELISA, BIO K302 (Bio-X Diagnostics, Belgium) and BIO K260 (Bio-X Diagnostics, Belgium). A cut-off of 135 AU was estimated to be optimal using Bayesian latent class analysis with three tests in multiple populations, accounting for conditional dependence between tests. At this cut-off, the DSe and DSp of the MilA ELISA were estimated to be 92.1 % (95 % highest probability density [HPD] interval: 87.4, 95.8) and 95.5 % (95 % HPD: 92.4, 97.8), respectively. The DSes of the BIO K260 and BIO K302 ELISAs were estimated to be 60.5 % (95 % HPD: 54.0, 66.9) and 44.6 % (95 % HPD: 38.7, 50.7), respectively. DSps were 95.6 % (95 % HPD: 92.9, 97.7) and 97.8 % (95 % HPD: 95.9, 99.0), respectively. Mycoplasma bovis seroprevalence was remarkably high at follow-up after 42 days on the feedlots. Overall, this study estimated a cut-off, DSe and DSp for the MilA ELISA with less dependence on prior information than previous analyses and demonstrated that the MilA ELISA has higher DSe than the BIO K260 and BIO K302 assays.

Background Drowning deaths occur among people of all ages, however, the risk is not uniform across the lifespan. Conventional drowning prevention programs communicate risk using age bands, however, this approach may overlook critical time points through the life cycle. Methods The Royal Life Saving National Fatal Drowning Database was used to examine drowning by single year of age from 1 July 2002 to 30 June 2019. This database includes all unintentional drowning deaths in Australian waters. Results Drowning peaked at one year of age, with a significant spike observed following the first birthday (0 years: 1.2/100,000 vs 1 year: 4.2/100,000). Deaths decreased as children grew, before rising sharply at 15–19 years of age (15 years: 0.5/100,000 vs 19 years: 1.6/100,000). The drowning rate remained steady through adulthood, before rising again in early retirement (60 years: 1.8/100,000). Conclusion Drowning risk changes across the lifespan, with the greatest risk observed among young children as they become increasingly mobile. A peak also occurs as teenagers reach adulthood, legal drinking age and gain greater independence. Assessing risk by single year of age allows for the design of targeted strategies, aligned to physical development stages and social changes. Learning Outcomes Drowning prevention strategies need to be tailored to specific life stages, taking into consideration changes in mobility and independence. For example, existing broad prevention strategies for children aged 0–4 could be further tailored to highlight the importance of supervision for newly mobile for 1–2 year olds and water safety education for 3–4 year olds.

Background Drowning is a leading cause of injury-related mortality and morbidity in Australia. In accordance with the Australian Water Safety Strategy (AWSS), advocacy efforts have been directed towards areas of high risk, underlined by an aspirational goal of a 50% reduction in drowning by 2020. Methods A total population analysis of unintentional, fatal drowning was conducted using the Royal Life Saving National Fatal Drowning Database. The study baseline (three-year average from 2004/05–2006/07) was compared to current figures (three-year average from 2016/17–2018/19), aligning with the beginning and end of the AWSS aspirational goal. Results Between 2004/05–2006/07 and 2016/17–2018/19, the overall drowning rate reduced by 20.5%. Substantial decreases were recorded against AWSS Goal 1 children 0–4 years (49.2% decrease) and 5–9 years (55.5% decrease), with negligible decreases or increases among people aged 75 years and over (Goal 4). The location-specific drowning rate at rivers decreased by 24.0% (Goal 5) and beaches by 5.5% (Goal 6) but increased at rocks by 35.3%. Seven of Australia’s eight States and Territories recorded reductions in the rate. Conclusion Australia’s drowning rate has reduced over the course of the AWSS, with significant progress made in children (Goal 1) but little to no success in reducing drowning among older people (Goal 4). Future iterations of the AWSS should prioritise areas of limited progress by developing and implementing targeted drowning prevention strategies. Learning Outcomes Future drowning prevention efforts should be directed towards areas of limited progress, while consideration needs to be given to changing demographic trends and emerging issues.

We present the genome sequences of macropodid alphaherpesviruses 2 and 4, two closely related pathogens of macropods. Both encoded 68 nonredundant open reading frames (ORFs) and share 90.6% genome-wide nucleotide identity. These viruses are associated with fatal outbreaks of disease in multiple marsupial species. These sequences will be important for the development of new diagnostic tools.

Adjuvants are formulated in conventional vaccines to ensure immune recognition and the subsequent development of innate and/or adaptive immune responses. A principal advantage of using a viral vector is that no adjuvant is required as the vector alone can achieve the desired goals of eliciting the required immune responses. However, in some cases, the viral vector may be attenuated to achieve the desired level of in vivo safety, or it may not stimulate the most appropriate responses for the pathogen of interest. Consequently, the incorporation of an adjuvant may be required to augment the immune responses to the viral vector of interest including any heterologous antigen(s) it may encode. This chapter will describe with examples the two broad classes, convention and molecular, of adjuvants that have been utilized to improve the performance of live viral vectors. Potential unintended consequences of the use of adjuvants and the strategies used to minimize these effects while maintaining activities will also be discussed.

Campylobacter spp. contaminated poultry products are strongly associated with foodborne illnesses worldwide. Development of effective management strategies to reduce contamination by Campylobacter spp. requires an improved understanding of the numerous factors that drive these contamination processes. Currently, chicken farms are using more free-range chicken meat production systems in response to consumer preferences. However, Campylobacter spp. colonization has rarely been investigated on free-range broiler farms. The present study investigated the temporal and environmental factors influencing Campylobacter spp. colonization of free-range broilers as well as potential sources and genetic diversity of Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) in commercial free-range broiler farms. Genetic linkages among the isolates were analyzed using flaA amplicon analysis. Campylobacter coli was first detected in fecal samples of a commercial free-range broiler flock on day 10 of rearing. Multiple genotypes of C. jejuni and C. coli were identified in this study. The farm environment was identified as a potential source of C. jejuni and C. coli colonization of free-range broilers. The dominant Campylobacter genotype varied between free-range broiler farms over time, with C. jejuni being the most frequently isolated species. These findings enhance the understanding of C. jejuni and C. coli colonization in free-range broiler farms and could inform the development of more effective intervention strategies to help control this important foodborne pathogen.

Viruses belonging to the Herpesviridae family are widely distributed in nature and are associated with many diseases. Herpesvirus infections and associated diseases of production animals are of particular importance due to economic losses incurred by industry and significant animal health and welfare issues. To address herpesvirus-associated diseases in production animals, the use of vaccination has been widely applied. To further enhance vaccine efficiency, concerted efforts to develop vaccination systems that provide optimal safety profiles while providing protective immunological responses is an area of intense study. This includes research to exploit herpesviruses as viral vector systems that can deliver heterologous antigens from other viral and bacterial pathogens as a multivalent viral vector system. This chapter describes the construction and development of several species-specific Alphaherpesvirinae-derived vector platforms which have been exploited as multivalent live viral vaccines against pathogens affecting horse, swine, poultry and cattle industries.

Amphibians are becoming increasingly reliant on captive breeding programs for continued survival. Assisted reproductive technologies including gamete cryopreservation and IVF can help reduce costs of breeding programs, provide insurance against extinction and assist genetic rescue in wild populations. However, the use of these technologies to produce reproductively mature offspring has only been demonstrated in a few non-model species. We aimed to optimise sperm cryopreservation in the threatened frog Litoria aurea and generate mature offspring from frozen-thawed spermatozoa by IVF. We tested three concentrations (1.4, 2.1 and 2.8M) of the cryoprotectants dimethylsulfoxide (DMSO) and glycerol with 0.3M sucrose. Using DMSO was more likely to result in recovery of sperm motility, vitality and acrosome integrity than glycerol, regardless of concentration, with forward progressive motility being most sensitive to damage. The lowest concentrations of 1.4 and 2.1M provided the best protection regardless of cryoprotectant type. Spermatozoa cryopreserved in 2.1M DMSO outperformed spermatozoa cryopreserved in equivalent concentrations of glycerol in terms of their ability to fertilise ova, resulting in higher rates of embryos hatching and several individuals reaching sexual maturity. We have demonstrated that sperm cryopreservation and subsequent offspring generation via IVF is a feasible conservation tool for L. aurea and other threatened amphibians.

Most fish species are susceptible to red-spotted grouper nervous necrosis virus (RGNNV) which causes viral encephalopathy and retinopathy leading to huge financial losses in the aquaculture farming industry for decades. A knowledge of RGNNV replication and the host response in depth is warranted to understand the host-virus interaction and to develop an effective approach to inhibit this virus. This study therefore aimed to investigate the expression of selected immune and stress-related genes of the E-11 clonal cell line upon RGNNV infection. The results showed significant upregulation of viperin, hsp70 and tnfaip3 transcripts at 48 h post infection, while the transcript abundance of irf3, cc chemokine, hsp30 showed increases which did not reach significance and tnf-α showed no change in expression in response to infection. This study highlights the involvement of viperin, hsp70 and tnfaip3 during RGNNV infection of E-11 cells which provides a clearer understanding of the pathogenic mechanism of the disease.

Animal health measures mainly rely on vaccination or chemical control for major pests and pathogens, causing issues of residue, toxicity and development of resistance. For example, control of Sheep flystrike and lice-infestation affecting the Australia’s sheep/wool industry (>3.5 B) have developed resistance to nearly all control chemicals used in the past. Topicals RNAi provides an innovative clean-green, non-toxic, environmentally sustainable biological control solution. Biodegradable clay particles as carriers can be used to deliver double stranded RNA (dsRNA), the key trigger molecule of RNA interference pathway. As an early proof of concept, we investigated the stability dsRNA loaded on two types of Clay particles: Clay 1 (releases dsRNA under acidic conditions) and Clay 2 (releases dsRNA under alkaline conditions) on cattle hide. Cattle skin was treated with Cy3 labelled dsRNA alone and Cy3 labelled dsRNA loaded on Clay1 or Clay2. The skin samples treated with the Cy3 formulations were imaged using confocal microscopy. Once imaged, the skin samples were washed and stored at room temperature for 5 days, later the samples were re-imaged to detect the fluorescent signal (Figure 1). The dsRNA loaded on clay particles was stable unlike naked Cy3-dsRNA which degraded and was not visible after washing. This increased inherent stability of the dsRNA molecules, combined with the environmental stability afforded by the Clay particles, offers promise to provide a sustainable solution for animal health. Topical RNAi can reduce reliance on trade withholding periods of meat/wool without chemical residues, enhance animal welfare and increase production of premium quality meat/wool, improve export potential, competitiveness and long-term profitability of livestock industry.

Mycoplasma bovis can be a bacterial inhabitant of the upper respiratory tract of healthy bovines. In body regions other that the upper respiratory tract however, M. bovis is associated with a number of clinical syndromes such as bovine respiratory disease (BRD). This study used two enzyme-linked immunosorbent assays to assess the sero-status of M. bovis-specific antibodies in Australian feeder cattle at the time of feedlot induction and at approximately 42 days on feed (follow-up). The apparent sero-prevalence of M. bovis-specific antibody at induction was estimated to be 3.5% (95% confidence interval [CI] 2.0-5.0%, 47/1354) and 25.3% (95% CI 21.9-28.8%, 343/1354) at follow-up. Exposure to M. bovis between induction and follow-up as demonstrated by an increase in serum antibodies was estimated to be 19.4% (95% CI 16.2-22.6%, 261/1349). Risk factors associated with sero-positivity at feedlot induction included the region where animals were 28 days prior to induction and saleyard exposure at least 27 days prior to induction. Risk factors associated with a sero-increase between induction and follow-up included breed, source region and access to water shared with an adjoining pen of animals. Of these, shared pen water was considered the most important (odds ratio [OR] 3.3, 95% CI 1.5-7.4, p = 0.003). Animals exposed to M. bovis between induction and follow-up were at a substantially increased risk of BRD (OR 2.2, 95% CI 1.4-3.4, p = 0.001). This is the first Australian study that has identified risk factors for M. bovis sero-positivity and sero-increase and shown an association between sero-increase and the risk of BRD in the feeder cattle population. These findings suggest that M. bovis is a significant pathogen in the Australian feeder cattle population. In addition, identification of defined risk factors associated with an increased risk of exposure to M. bovis can assist in the development of targeted control measures to reduce the economic impact of M. bovis associated disease and BRD in feeder cattle.

Results obtained from a nationwide longitudinal study were extended to estimate the population-level effects of selected risk factors on the incidence of bovine respiratory disease (BRD) during the first 50days at risk in medium-sized to large Australian feedlots. Population attributable fractions (PAF) and population attributable risks (PAR) were used to rank selected risk factors in order of importance from the perspective of the Australian feedlot industry within two mutually exclusive categories: 'intervention' risk factors had practical strategies that feedlot managers could implement to avoid exposure of cattle to adverse levels of the risk factor and a precise estimate of the population-level effect while 'others' did not. An alternative method was also used to quantify the expected effects of simultaneously preventing exposure to multiple management-related factors whilst not changing exposure to factors that were more difficult to modify. The most important 'intervention' risk factors were shared pen water (PAF: 0.70, 95% credible interval: 0.45-0.83), breed (PAF: 0.67, 95% credible interval: 0.54-0.77), the animal's prior lifetime history of mixing with cattle from other herds (PAF: 0.53, 95% credible interval: 0.30-0.69), timing of the animal's move to the vicinity of the feedlot (PAF: 0.45, 95% credible interval: 0.17-0.68), the presence of Bovine viral diarrhoea virus 1 (BVDV-1) in the animal's cohort (PAF: 0.30, 95% credible interval: 0.04-0.50), the number of study animals in the animal's group 13days before induction (PAF: 0.30, 95% credible interval: 0.10-0.44) and induction weight (PAF: 0.16, 95% credible interval: 0.09-0.23). Other important risk factors identified and prioritised for further research were feedlot region, season of induction and cohort formation patterns. An estimated 82% of BRD incidence was attributable to management-related risk factors, whereby the lowest risk category of a composite management-related variable comprised animals in the lowest risk category of at least four of the five component variables (shared pen water, mixing, move timing, BVDV-1 in the cohort and the number of animals in the animal's group-13). This indicated that widespread adoption of appropriate interventions including ensuring pen water is not shared between pens, optimising animal mixing before induction, timing of the animal's move to the vicinity of the feedlot, and group size prior to placing animals in feedlot pens, and avoiding BVDV-1 in cohorts could markedly reduce the incidence of BRD in medium-sized to large Australian feedlots.

Aim To investigate if there are any trends in the age at diagnosis of Inflammatory Bowel Disease (IBD) patients who attend the Paediatric Department at University Hospital Limerick. Method Using our database of IBD patients, we analysed the charts of those diagnosed between Jan 1st2014 and Dec 31st2018. We extracted their date of birth, date of diagnosis, whether they were diagnosed with Ulcerative Colitis (UC) or Crohn’s Disease (CD) and tabulated the results using Excel. Results Over the time-period studied, in total 41 children and adolescents were diagnosed with IBD (59% CD, 41% UC). After stratifying by year there was no clear increase or decrease in the number of diagnoses per year from 2014 to 2018. The average age at diagnosis of IBD across this time period was 10.7 years (Range 2–16 years) and the average number of patients diagnosed per year was 8 (Range 5–11 patients per year). When looking separately at those diagnosed with CD, the average age of diagnosis across our 5-year time period was 10.48y (Range 2y-16y). When looking at those diagnosed with UC, the average age of diagnosis across our 5-year time period was 11.73y (Range 4y-16y) We analysed the age at diagnosis of both CD and UC by year in order to assess if there were any trends in age of diagnosis. For CD, the average age of diagnoses are as follows: 2014; 10y (range 7y to 12y), 2015; 11.4y (range 9y to 15y), 2016; 13y (range 11y to 15y), 2017; 11.25y (4y to 16y), 2018 6.75y (2y to 11y). For UC, the average age of diagnoses are as follows; 2014; 10y (range 9y to 12y), 2015 9.5y (4y to 13y), 2016; 12.67y (range 11y to 14y), 2017; 14y (13y to 15y), 2019; 11.73y (8y to 15y). We then looked at trends in ‘Very Early Onset IBD’ (VEO IBD) patients, which are classified as those Discussion The average age at diagnosis remains relatively unchanged across the years with no obvious trend in either direction, however when analysing the subgroup of VEO IBD diagnoses, it is clear that for CD specifically there has been an increase in the number of children diagnosed under the age of 6, with there being zero VEO IBD diagnoses from 2014–2016 and two per year in 2017 and 2018. Reference Snapper SB. Very-Early-Onset Inflammatory Bowel Disease. Gastroenterol Hepatol (N Y). 2015;11(8):554–6.

Aim The author describes a case of cholelithiasis occurring in an 18 month old female who was exposed to intravenous ceftriaxone for 5 days. Four weeks after ceftriaxone therapy, all but one stone had resolved. Case report An 18 month old female presented to the emergency department with high grade pyrexia and rigors. Physical examination and investigations did not reveal a focus for infection. Bloods: White Cell Count 46.6, Neutrophils 41.7 CRP 120. The patient was commenced on ceftriaxone as empirical treatment. Intermittent high grade pyrexia continued for 5 days. On day 5 an abdominal ultrasound was performed which demonstrated multiple gallbladder calculi within a normal thickness gallbladder. The appearance of the common bile duct was normal. The patient was discharged on day 7 having been apyrexial for 48 hours and clinically well. The patient re-presented 4 weeks later with high grade pyrexia. The patient was commenced on IV augmentin at presentation; gentamicin was added on day 3. A repeat ultrasound was performed and demonstrated resolution of all but one echogenic focus in the gallbladder. Ulstrasound of the abdomen at 8 weeks demonstrated complete resolution of the calculi. Discussion Cholelithiasis and choledocholithiasis were considered to be uncommon in infants and children but have been increasingly diagnosed in recent years due to the wide- spread use of ultrasonography.1 Ceftriaxone induced pseudolithiasis is an uncommon but recognized side effect of therapy and tends to resolve with cessation of therapy.2 In the paediatric population, where gallstones are uncommon, paediatricians and radiologists need to be aware of this association in order to avoid unnecessary intervention.3 References Poddar U. Gallstone disease in children. Indian Pediatr. 2010 Nov;47;11:945–53. Schaad UB, Wedgewood-Krucko J, Tschaeppeler H. Reversible Ceftriaxone associated pseudolithiasis in children. 1988;2: 1411–13. Nayak A, Slivka A. Ceftriaxone- Induced Gallstones: Case Report and Literature Review. ACG Case Rep J 2014:1:170–172.

Introduction Discitis – intervertebral disk infection although uncommon in children can pose a diagnostic challenge in uncommunicative toddlers. It may present with refusal to crawl or walk; limp or back pain, lumbar lordosis or a combination of one or more of these symptoms. We present a case of a delayed diagnosis of discitis in a toddler. Case presentation A 2 year old girl was referred to the emergency department for the second time in 10 days with a 3 week history of left hip pain and limp without a fever. She had a history of preceding viral URTI 2 weeks prior to her symptoms. The child’s mother thought the problem was the child’s ‘back’. On initial examination, she was noted to be well but limping/very uncomfortable walking and had a full range of movement of both hips. Initial impression was transient synovitis. Inflammatory markers increased over 48 hours with no symptom improvement on conservative management. MRI spine done on day 6 of admission showed an L5-S1 discitis osteomyelitis involving the sacral ala with small epidural abscess formation. She was subsequently treated on a prolonged course of intravenous antibiotics with complete resolution of symptoms. Conclusion Discitis is a difficult diagnosis to make in young children who are unable to describe their symptoms. It is important to listen to parental concerns on their own observations. As most cases of transient synovitis occur in children 3–10 years and typically resolve by 7–10 days, it is important to consider other diagnosis in a younger child with persistent symptoms to avoid unnecessary delay in treatment.

When compared with other age groups, fatal drowning rates among seniors have stayed static. This study identifies causal factors in unintentional bathtub drowning deaths among people aged 65 years and over. This study is a 10-year (2003-2012) total population retrospective survey of all unintentional bathtub (baths, spa baths and showers) drowning deaths afflicting people aged 65 years and over. Data were sourced from the Australian National Coronial Information System. Risk factors and circumstantial variables were analysed including sex, age, activity prior to drowning, alcohol, drugs, pre-existing medical conditions, living circumstances, time until found, and performance of cardio-pulmonary resuscitation (CPR). There were 32 fatalities (20 females, 12 males); 41% drowned after a fall into the bath. High blood alcohol (≥0.05%) was disproportionately represented. Twenty-six people (81%) had a pre-existing medical condition, deemed contributory in 19 cases including cardiac (n=9) and sarcopenia or frailty (n=5). Of those with medical conditions, 69% had blood levels of prescribed drugs, commonly analgesics (n=10). Seven cases recorded both drugs and alcohol. In half of all fatalities (50%), the person resided alone. In ten cases (31%) the person was not found for one or more days. Bathtub drowning afflicting the elderly poses an unmet challenge. This study has identified five areas for targeted prevention: Pre-existing medical conditions, alcohol, falls in the bath, review of medications; and if practical, advising family members of bathing. Increased awareness of drowning among this age group (and carers) is required as the aged population increases.

BackgroundShiga toxigenic Escherichia coli (STEC) is an emerging global pathogen and remains a major cause of food-borne illness with more severe symptoms including hemorrhagic colitis and hemolytic-uremic syndrome. Since the characterization of the archetypal STEC serotype, E. coli O157:H7, more than 250 STEC serotypes have been defined. Many of these non-O157 STEC are associated with clinical cases of equal severity as O157. In this study, we utilize whole genome sequencing of 44 STEC strains from eight serogroups associated with human infection to establish their evolutionary relationships and contrast this with their virulence gene profiles and established typing methods.ResultsOur phylogenomic analysis delineated these STEC strains into seven distinct lineages, each with a characteristic repertoire of virulence factors. Some lineages included commensal or other E. coli pathotypes. Multiple independent acquisitions of the Locus for Enterocyte Effacement were identified, each associated with a distinct repertoire of effector genes. Lineages were inconsistent with O-antigen typing in several instances, consistent with lateral gene transfer within the O-antigen locus. STEC lineages could be defined by the conservation of clustered regularly interspaced short palindromic repeats (CRISPRs), however, no CRISPR profile could differentiate STEC from other E. coli strains. Six genomic regions (ranging from 500 bp - 10 kbp) were found to be conserved across all STEC in this dataset and may dictate interactions with Stx phage lysogeny.ConclusionsThe genomic analyses reported here present non-O157 STEC as a diverse group of pathogenic E. coli emerging from multiple lineages that independently acquired mobile genetic elements that promote pathogenesis.

Case reportA captive breeding colony of 9 greater bilbies (Macrotis lagotis) exhibited mild upper respiratory signs and sudden deaths with 100% mortality over a 2-week period. Histologically, acute necrotising and erosive epithelial lesions throughout the upper respiratory system and bronchi were associated with eosinophilic intranuclear inclusion bodies. Inclusions were also present in hepatocytes and adrenocortical cells, but were not always associated with necrosis. Transmission electron microscopy of lung sections revealed nucleocapsids forming arrays within some nuclei. A pan-herpesvirus PCR yielded a 440-bp product, with sequencing confirming homology with the alphaherpesviruses. Viral culture in a marsupial cell line resulted in cytopathic effect consistent with an alphaherpesvirus.