Metamorphosis Imposes Variable Constraints on Genome Expansion through Effects on Development.

Genome size varies ∼100,000-fold across eukaryotes and has long been hypothesized to be influenced by metamorphosis in animals. Transposable element accumulation has been identified as a major driver of increase, but the nature of constraints limiting the size of genomes has remained unclear, even as traits such as cell size and rate of development co-vary strongly with genome size. Salamanders, which possess diverse metamorphic and non-metamorphic life histories, join the lungfish in having the largest vertebrate genomes-3 to 40 times that of humans-as well as the largest range of variation in genome size. We tested 13 biologically-inspired hypotheses exploring how the form of metamorphosis imposes varying constraints on genome expansion in a broadly representative phylogeny containing 118 species of salamanders. We show that metamorphosis during which animals undergo the most extensive and synchronous remodeling imposes the most severe constraint against genome expansion, with the severity of constraint decreasing with reduced extent and synchronicity of remodeling. More generally, our work demonstrates the potential for broader interpretation of phylogenetic comparative analysis in exploring the balance of multiple evolutionary pressures shaping phenotypic evolution.

Haplosporidium acetes n. sp. infecting the hepatopancreas of jelly prawns Acetes sibogae australis from Moreton Bay, Australia.

Wild Acetes sibogae australis from northern Moreton Bay, Australia displaying opacity of the hepatopancreas were sampled and examined histologically, revealing infection by multinucleate plasmodia of a haplosporidian-like parasite in the epithelial cells of the hepatopancreas. A morphological and phylogenetic investigation identified the parasite as a novel species of the order Haplosporida, and the parasite is described as Haplosporidium acetes n. sp. This is the first report of disease caused by a haplosporidian in wild Australian decapod crustaceans, and the first record of haplosporidiosis in sergestid shrimp. Infections of H. acetes were observed in all cell types (R, B, F and E) within the hepatopancreas. Infected epithelial cells became hypertrophied as they filled with haplosporidian parasites and, in heavy infections, caused almost complete displacement of normal hepatopancreas tissue. Although sporulation was not observed, infected jelly prawns appeared terminally diseased. Infections became grossly evident in around 5% of wild prawns during early autumn at a time of year when jelly prawn populations decline rapidly with decreasing water temperatures, however histopathology indicated at least 13% of apparently normal jelly prawns were also infected. Further studies are required in order to determine if this parasite influences jelly prawn population dynamics. In addition, we report co-infection of a novel microsporidian parasite in the Enterocytozoon Group Microsporidia (EGM) infecting nuclei of hepatopancreatic epithelial cells. The microsporidian was phylogenetically distinct from Enterocytozoon hepatopenaei (EHP) known to infect penaeid shrimp in Asia.

A multi-site study on the impact of an advance care planning workshop on attitudes, beliefs and behavioural intentions over a 6-month period.

BACKGROUND: This study evaluated the impact of the adapted version of the Respecting Choices® The Living Matters Advance Care Planning (ACP) facilitator training programme on trainees' attitudes on facilitation 6 months post-training. SETTING AND PARTICIPANTS: Two hundred and twenty-one healthcare professionals consisting of doctors, nurses, medical social workers from different training venues in Singapore participated in the first phase of the study (pre- and post) of which 107 participated in the second phase 6 months later (follow-up). METHODS: Participants self-rated their attitudes, beliefs and behavioural intentions through surveys at three time points in an evaluation design that utilised repeated measures one-way ANOVA (pre-, post-, follow-up). Between-group differences were also examined using independent t-test. RESULTS: At follow-up, mean scores increased significantly in understanding, confidence, and competence. Changes in effect sizes were large. Although trainees continued to think that ACP is emotionally draining for facilitators, more than before, facilitation experience was considered pleasant for themselves with the positive change significant and moderate in effect size. Those who had experience completing/initiating ACP significantly held more positive views than those who did not. CONCLUSIONS: The ACP facilitator training programme had lasting effects on enhancing the understanding, competence, and confidence of trainees. Importantly, findings showed that experience in actual facilitation within 6 months after training was important and giving trainees opportunities to facilitate is recommended.

Circular RNA circ-SMAD7 is downregulated in colorectal cancer and suppresses tumor metastasis by regulating epithelial mesenchymal transition.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Circular RNA circ-SMAD7 is downregulated in colorectal cancer and suppresses tumor metastasis by regulating epithelial mesenchymal transition, by D.-K. Wang, R.-F. Chong, B.-L. Song, K.-F. Fan, Y.-F. Liu, published in Eur Rev Med Pharmacol Sci 2020; 24 (4): 1736-1742-DOI: 10.26355/eurrev_202002_20350-PMID: 32141541" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20350.

Circular RNA circ-SMAD7 is downregulated in colorectal cancer and suppresses tumor metastasis by regulating epithelial mesenchymal transition.

OBJECTIVE: Recently, circular RNAs play a vital role in many diseases including tumor progression. Colorectal cancer (CRC) is one of the most ordinary malignant tumors. The purpose of our study is to detect the potential function of circ-SMAD7 in CRC. PATIENTS AND METHODS: The level of circ-SMAD7 was detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) in CRC tissue samples. The circ-SMAD7 expression level and the patients' overall survival time were analyzed. Functional experiments were conducted to identify the changes of the biological behaviors in CRC cells after the overexpression of circ-SMAD7. The transwell assay, the Matrigel assay, and the Wound healing assay were conducted. The Western blot assay was performed to analyze the effect of circ-SMAD7 on the epithelial-to-mesenchymal transition (EMT) process. RESULTS: In the research, the expression level of circ-SMAD7 was significantly decreased in CRC tissues compared with that in the adjacent samples. Circ-SMAD7 expression was positively associated to patients' overall survival time. The expression of circ-SMAD7 was also decreased in CRC cell lines. The upregulation of circ-SMAD7 led to the inhibition of cell migration and invasion in CRC. In addition, the results of further experiments revealed that the EMT-related proteins were regulated via overexpression of circ-SMAD7 in CRC. CONCLUSIONS: These results suggest that circ-SMAD7 could inhibit cell migration and invasion of CRC by suppressing the EMT process, which might offer a potential therapeutic target for CRC.

Induction of M2 Macrophages Prevents Bone Loss in Murine Periodontitis Models.

Periodontitis is characterized by the progressive destruction of tooth-supporting alveolar bone, which is mainly caused by chronic inflammation in response to persistent bacterial insult. It has recently become clear that the pathogenesis of periodontitis is associated with a high ratio of proinflammatory M1 (classically activated) macrophages to anti-inflammatory M2 (alternatively activated). To decrease the inflammatory activity, we locally delivered the C-C motif chemokine ligand 2 (CCL2) using controlled-release microparticles (MPs). CCL2 is known to promote chemotaxis of M0 or M2 phenotype macrophages to the inflamed site and induce M2 phenotype polarization locally. Our in vitro data showed that CCL2 increased the number of M2 phenotype macrophages, decreased TNF-α secretion, and enhanced chemotaxis of RAW264.7 cells toward CCL2 MPs. Moreover, we induced periodontal disease in 2 animal models through inoculation of Porphyromonas gingivalis and ligature around the murine molar. Micro-computed tomography analysis showed significant reduction of alveolar bone loss in the CCL2 MP treatment group when compared with a blank MP group and a no-treatment periodontitis group in both models. Immunohistologic analysis showed a significant increase in the M2 phenotype subset and a decrease in the M1 phenotype subset in the CCL2 MP group of the P. gingivalis-induced model. Also, in both models, tartrate-resistant acidic phosphatase staining showed significantly fewer numbers of osteoclasts in the CCL2 MP group in alveolar bone area. Moreover, quantitative polymerase chain reaction results showed a significant increase in IL-1RA (interleukin 1 receptor antagonist) mRNA expression and a decrease in RANKL (receptor activator of nuclear factor kappa-Β ligand) mRNA expression in the CCL2 MP group in the ligature model. In summary, manipulation of endogenous M2 phenotype macrophages with CCL2 MPs decreased the M1 phenotype:M2 phenotype ratio and prevented alveolar bone loss in mouse periodontitis models. The delivery of CCL2 MPs provides a novel approach to treat periodontal disease.

Establishment of institutional diagnostic reference level for computed tomography with automated dose-tracking software.

INTRODUCTION: The aim of this study was to establish institutional diagnostic reference levels (DRLs) by summarising doses collected across the five computed tomography (CT) system in our institution. METHODS: CT dose data of 15940 patients were collected retrospectively from May 2015 to October 2015 in five institutional scanners. The mean, 75th percentile and 90th percentile of the dose spread were calculated according to anatomic region. The common CT examinations such as head, chest, combined abdomen/pelvis (A/P), and combined chest/abdomen/pelvis (C/A/P) were reviewed. Distribution of CT dose index (CTDIvol), dose-length product (DLP) and effective dose (ED) were extracted from the data for single-phasic and multiphasic examinations. RESULTS: The institutional DRL for our CT units were established as mean (50th percentile) of CTDIvol (mGy), DLP (mGy.cm) and ED (mSv) for single and multiphasic studies using the dose-tracking software. In single phasic examination, Head: (49.0 mGy), (978.0 mGy.cm), (2.4 mSv) respectively; Chest: (6.0 mGy), (254.0 mGy.cm), (4.9 mSv) respectively; CT A/P (10.0 mGy), (514.0 mGy.cm), (8.9 mSv) respectively; CT C/A/P (10.0 mGy), (674.0 mGy.cm), (11.8 mSv) respectively. In multiphasic studies: Head (45.0 mGy), (1822.0 mGy.cm), (5.0 mSv) respectively; Chest (8.0 mGy), (577.0 mGy.cm), (10.0 mSv) respectively; CT A/P: (10.0 mGy), (1153.0 mGy.cm), (20.2 mSv) respectively; CT C/A/P: (11.0 mGy), (1090.0 mGy.cm), (19.2 mSv) respectively. CONCLUSIONS: The reported metrics offer a variety of information that institutions can use for quality improvement activities. The variations in dose between scanners suggest a large potential for optimisation of radiation dose.

Measurement of serum 3-epi-25-hydroxyvitamin D3, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 in infant, paediatric and adolescent populations of Korea using ultra-performance liquid chromatography-tandem mass spectrometry.

Background We evaluated the performance of ultra-performance liquid chromatography-tandem mass spectrometry to measure serum 3-epi-25-hydroxyvitamin D3, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 concentrations in 519 infant, paediatric and adolescent serum samples in Korea. Methods We used a Kinetex XB-C18 column and isocratic methanol/water (77.5/22.5, v/v) with 0.025% (v/v) high-performance liquid chromatography solvent additive flowing at 0.25 mL/min, yielding an 11 min/sample run time. A TQD triple quadrupole mass spectrometer in electrospray ionization positive ion mode with multiple reaction monitoring transition via an MSMS vitamin D kit was used to evaluate precision, carryover, ion suppression and linearity. Samples were prepared using the 4-phenyl-1,2,4-triazoline-3,5-dione derivatization method. Results Intra- and inter-run precisions were 1.23-13.28% and 1.02-10.08%, respectively. Group carryovers were -0.27% and 0.10%, respectively. There was no ion suppression. The calibration curve showed good linearity from calibrator Level 1 (11.75 nmol/L) to 6 (375 nmol/L) with R2 > 0.9999. The 3-epi-25-hydroxyvitamin D3 and 25-hydroxyvitamin D3 peaks were clearly separated in the extracted ion chromatogram. Infant serum samples 3-epi-25-hydroxyvitamin D3 concentrations were significantly higher than paediatric and adolescent concentrations. Conclusions The ultra-performance liquid chromatography-tandem mass spectrometry assay performed acceptably, clearly separating 3-epi-25-hydroxyvitamin D3 from 25-hydroxyvitamin D3. High 3-epi-25-hydroxyvitamin D3 concentrations were observed in infant but not in paediatric and adolescent serum samples.

Individuals with a vestibular-related disorder use a somatosensory-dominant strategy for postural orientation after inclined stance.

OBJECTIVES: The visual, somatosensory, and vestibular systems are critical for establishing a sensorimotor set for postural control and orientation. The goal of this study was to assess how individuals with a vestibular-related disorder keep their balance following prolonged stance on an inclined surface. We hypothesize that subjects will show greater reliance on the somatosensory system than age-matched controls as inferred by the presence of a forward postural lean aftereffect following the inclined stance (i.e., a positive response). RESULTS: The results revealed an underlying somatosensory-dominant strategy for postural control in the vestibular group: 100% of the subjects tested positive compared to 58% in the control group (P=.006). CONCLUSION: Individuals with a vestibular-related disorder use a somatosensory-dominant strategy for postural orientation following prolonged inclined stance. The implications for the management of this population are discussed.

Detection of infectious spleen and kidney necrosis virus (ISKNV) and turbot reddish body iridovirus (TRBIV) from archival ornamental fish samples.

Although infections caused by megalocytiviruses have been reported from a wide range of finfish species for several decades, molecular characterisation of the viruses involved has been undertaken only on more recent cases. Sequence analysis of the major capsid protein and adenosine triphosphatase genes is reported here from formalin-fixed, paraffin-embedded material from 2 archival ornamental fish cases from 1986 and 1988 in conjunction with data for a range of genes from fresh frozen tissues from 5 cases obtained from 1991 through to 2010. Turbot reddish body iridovirus (TRBIV) genotype megalocytiviruses, previously not documented in ornamental fish, were detected in samples from 1986, 1988 and 1991. In contrast, megalocytiviruses from 1996 onwards, including those characterised from 2002, 2006 and 2010 in this study, were almost indistinguishable from infectious spleen and kidney necrosis virus (ISKNV). Three of the species infected with TRBIV-like megalocytiviruses from 1986 to 1991, viz. dwarf gourami Trichogaster lalius (formerly Colisa lalia), freshwater angelfish Pterophyllum scalare and oscar Astronotus ocellatus, were infected with ISKNV genotype megalocytiviruses from 2002 to 2010. The detection of a TRBIV genotype isolate in ornamental fish from 1986 represents the index case, confirmed by molecular sequence data, for the genus Megalocytivirus.

Metformin Has a Positive Therapeutic Effect on Prostate Cancer in Patients With Type 2 Diabetes Mellitus.

OBJECTIVE: Prostate cancer and type 2 diabetes mellitus (DM2) are both common diseases found in the elderly male population. The diabetic drug, metformin, has been shown to have antineoplastic properties and demonstrated better treatment outcomes when used as adjuvant therapy in patients with breast cancer. The hormonally-sensitive cancer analogous to breast cancer in men is prostate cancer. We investigated improved survival, lower risks of recurrences, and lower, more stable levels of prostate-specific antigen (PSA) in patients with DM2 along with prostate cancer on metformin. METHODS: Patients with prostate cancer along with DM2 who remained on metformin were compared with controls who were not on metformin matched by age, weight, race and Gleason score cancer staging. The endpoints of our study included final PSA values, number of recurrences, metastases and number living for each group. RESULTS: There were significantly fewer deaths (23% versus 10%), fewer recurrences (15% versus 8%), fewer metastases (5% versus 0%) and fewer secondary cancers (17% versus 6%) in the metformin group (P < 0.004). The final PSA value was lower in the metformin-treated group with a result approaching significance (P = 0.067). The primary treatments for prostate cancer (ie, surgery, radiation and androgen depletion) were found to be comparable in both the groups. CONCLUSIONS: Our retrospective study shows that adjuvant metformin therapy leads to a better prognosis in prostate cancer. Not only are PSA levels controlled for several years but also there are significantly fewer cancer recurrences in metformin-treated patients. Overall, these results are promising and should be followed up with a prospective study to assess long-term survival.

Comparative gait initiation kinematics between simulated unilateral and bilateral ankle hypomobility: Does bilateral constraint improve speed performance?

Improvement of motor performance in unilateral upper limb motor disability has been shown when utilizing inter-limb coupling strategies during physical rehabilitation. This suggests that 'default' bilateral central motor commands are facilitated. Here, we tested whether this bilateral motor control principle may be generalized to the lower limbs during gait initiation, which involves alternate bilateral actions. Disability was simulated by strapping to produce ankle hypomobility. Healthy adult subjects initiated gait at a self-paced speed with no ankle constraint (control), or with the stance, swing or bilateral ankles strapped. The duration of the anticipatory postural adjustments lengthened and the center of mass instantaneous progression velocity at foot-off decreased when the ankle was strapped. During the step execution phase, progression velocity at foot-contact was higher when both ankles were strapped compared to unilateral strapping of the stance ankle. These findings suggest that bilateral central motor commands are favored during walking tasks. Indeed, unilateral constraint of the stance ankle should compel the central nervous system to adapt specific commands to the constraint and normal sides whereas the 'default' bilateral motor commands would be utilized when both ankles are strapped leading to better kinematics performance. Bilateral in-phase upper limb coordination and bilateral alternating lower limb locomotor movements may share similar control mechanisms.

Duration of step initiation predicts freezing in Parkinson's disease.

OBJECTIVES: In some individuals with idiopathic Parkinson's disease (PD), freezing of gait episodes develops as the disease progresses. The neural mechanism underlying freezing in PD is poorly understood. Here, we report a 2-year follow-up on the novel discovery of prolonged step initiation duration as a potential marker of impending freezing. METHODS: Non-freezing PD participants in stages 2.5-4 of the Hoehn and Yahr disease severity scale were recruited from an earlier study which determined the effect of semi-virtual cues on walking. Responders were those who completed the first step faster in the presence of the virtual cues while non-responders either did not change or took longer to complete the first step. Both groups of participants were interviewed 2 years later to determine who had developed freezing of gait. RESULTS: Participants in the responder group had a 13-fold risk of developing freezing of gait within 2 years following the cueing study (OR=13.3, 95% CI=1.1-167). A cutoff score of -2.6% (i.e., a decrease in the duration of the first step with visual cues by 2.6% relative to no cues) gave a sensitivity and specificity of 100% and 89%, respectively. CONCLUSIONS: To the best of our knowledge, this is the first novel discovery of a physical predictor of freezing in PD. The time to complete the first step is a simple test to administer in the clinic or at home and may therefore be easily incorporated into a fall prevention training program for PD before the inception of freezing.

Diagnostic value of the rapid assessment of postural instability in Parkinson's disease (RAPID) questionnaire.

BACKGROUND: People with idiopathic Parkinson's disease (PD) develop postural instability in the later stages of the ailment. Postural instability has traditionally been quantified with the Pull test even though its face validity is limited. We previously established cut-off scores for a three-part rapid assessment of postural instability (RAPID) questionnaire as a non-physical complement to the physical test. In the current study, the questionnaire was administered to a new group of PD subjects to evaluate the diagnostic value of the instrument. METHODS: Sensitivity and specificity values were calculated for single and combined sections of the questionnaire by using the Pull test as the gold standard for assessing the presence of postural instability. RESULTS: The questionnaire when used in its entirety gave the highest sensitivity (.71), whereas specificity was highest in the activities of daily living (.74) and fear of falling sections (.74). Net specificity decreased to .44 when the scores from the three sections of the questionnaire were combined. CONCLUSIONS: The high sensitivity of the RAPID questionnaire suggests that it may be used as an adjunct to the Pull test or solely if it is not convenient or contraindicated. The questionnaire may also be adapted for use via the telephone or internet. The limitation of the Pull test in revealing postural instability may explain the low specificity of the questionnaire, i.e. the questionnaire correctly identifies patients as unstable when the Pull test indicates normal postural control. It is hoped that the rapid identification of postural instability in PD may lead to increased awareness of the disease progression and fewer falls.

Ministry of Health clinical practice guidelines: Chronic Hepatitis B Infection.

The Ministry of Health (MOH) publishes clinical practice guidelines on Chronic Hepatitis B Infection to provide doctors and patients in Singapore with evidence-based guidance on managing important medical conditions. This article reproduces the introduction and executive summary (with recommendations from the guidelines) from the MOH clinical practice guidelines on Chronic Hepatitis B Infection, for the information of readers of the Singapore Medical Journal. Chapters and page numbers mentioned in the reproduced extract refer to the full text of the guidelines, which are available from the Ministry of Health website (http://www.moh.gov.sg/mohcorp/publications.aspx?id=26108). The recommendations should be used with reference to the full text of the guidelines. Following this article are multiple choice questions based on the full text of the guidelines.

Rapid assessment of postural instability in Parkinson's disease (RAPID): a pilot study.

BACKGROUND: The Fahn's pull (or retropulsion) test is an item in the motor section of the Unified Parkinson's Disease Rating Scale, which is used almost exclusively to classify postural instability in Parkinson's disease (PD). However, the test is hard to standardize and is often performed incorrectly, making it hard to interpret. Moreover, it may not be safe to administer in patients who experience pain in the shoulders, neck, trunk and/or lower extremities. Identifying and grading postural instability in PD without requiring a physical challenge would not only be useful for the clinician but would assist patients and caregivers in its recognition. We propose the use of the rapid assessment of postural instability in Parkinson's disease (RAPID) questionnaire as a non-physical assessment tool. METHODS: We determined the associations between the pull test and items on a risk-assessment questionnaire that consisted of three parts: activities of daily living, fear of falling, and frequency of falling. RESULTS: Significant correlations were found between the pull test and the predictor variables, which ranged between 0.51 and 0.56 whilst the correlations amongst the predictor variables ranged between 0.58 and 0.70. The three parts of the questionnaire, when used in combination, produced a 96% sensitivity in the classification of postural instability. CONCLUSIONS: The RAPID questionnaire can be used as an adjunct to the pull test or solely if the pull test is contraindicated. It may also be possible to administer the questionnaire via the telephone or Internet. It is hoped that the rapid identification of postural instability would lead to fewer falls.

Influence of sensory inputs and motor demands on the control of the centre of mass velocity during gait initiation in humans.

Human gait requires the simultaneous generation of goal-directed continuous movement (locomotion) and the maintenance of balance (postural control). In adults, the centre of mass (CoM) oscillates in the vertical plane while walking. During the single support phase of gait initiation, its vertical (vCoM) velocity increases as the CoM falls and is actively reversed prior to foot-contact. In this study we investigated whether this active control, which is thought to reflect balance control during gait initiation, is controlled by visual and somatosensory inputs (Experiment 1) and whether it is modified by a change in motor demands, two steps versus one step (Experiment 2). In all healthy adults, the vCoM velocity was braked, or controlled, by contraction of the soleus muscle of the stance leg. The elimination of visual input alone had no effect on braking, although its amplitude decreased when somatosensory inputs were disrupted (-47%), and further decreased when both visual and somatosensory inputs were disrupted (-83%). When subjects performed only one step, with no trailing of the stance foot, the vCoM velocity braking also decreased (-42%). These results suggest that active braking of the CoM fall during the transition to double support, an indicator of balance control, is influenced by both multisensory integration and the demands of the current motor program. The neural structures involved in this mechanism remain to be elucidated.

Global trade in ornamental fish from an Australian perspective: the case for revised import risk analysis and management strategies.

Over 1 billion ornamental fish comprising more than 4000 freshwater and 1400 marine species are traded internationally each year, with 8-10 million imported into Australia alone. Compared to other commodities, the pathogens and disease translocation risks associated with this pattern of trade have been poorly documented. The aim of this study was to conduct an appraisal of the effectiveness of risk analysis and quarantine controls as they are applied according to the Sanitary and Phytosanitary (SPS) agreement in Australia. Ornamental fish originate from about 100 countries and hazards are mostly unknown; since 2000 there have been 16-fold fewer scientific publications on ornamental fish disease compared to farmed fish disease, and 470 fewer compared to disease in terrestrial species (cattle). The import quarantine policies of a range of countries were reviewed and classified as stringent or non-stringent based on the levels of pre-border and border controls. Australia has a stringent policy which includes pre-border health certification and a mandatory quarantine period at border of 1-3 weeks in registered quarantine premises supervised by government quarantine staff. Despite these measures there have been many disease incursions as well as establishment of significant exotic viral, bacterial, fungal, protozoal and metazoan pathogens from ornamental fish in farmed native Australian fish and free-living introduced species. Recent examples include Megalocytivirus and Aeromonas salmonicida atypical strain. In 2006, there were 22 species of alien ornamental fish with established breeding populations in waterways in Australia and freshwater plants and molluscs have also been introduced, proving a direct transmission pathway for establishment of pathogens in native fish species. Australia's stringent quarantine policies for imported ornamental fish are based on import risk analysis under the SPS agreement but have not provided an acceptable level of protection (ALOP) consistent with government objectives to prevent introduction of pests and diseases, promote development of future aquaculture industries or maintain biodiversity. It is concluded that the risk analysis process described by the Office International des Epizooties under the SPS agreement cannot be used in a meaningful way for current patterns of ornamental fish trade. Transboundary disease incursions will continue and exotic pathogens will become established in new regions as a result of the ornamental fish trade, and this will be an international phenomenon. Ornamental fish represent a special case in live animal trade where OIE guidelines for risk analysis need to be revised. Alternatively, for countries such as Australia with implied very high ALOP, the number of species traded and the number of sources permitted need to be dramatically reduced to facilitate hazard identification, risk assessment and import quarantine controls.