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Background: Knowledge of post-myocardial infarction (MI) disease risk to date is limited-yet the number of survivors of MI has increased dramatically in recent decades. We investigated temporally ordered sequences of all conditions following MI in nationwide electronic health record data through the application of process mining., Methods: We conducted a national retrospective cohort study of all hospitalisations (145,670,448 episodes; 34,083,204 individuals) admitted to NHS hospitals in England (1st January 2008-31st January 2017, final follow-up 27th March 2017). Through process mining, we identified trajectories of all major disease diagnoses following MI and compared their relative risk (RR) and all-cause mortality hazard ratios (HR) to a risk-set matched non-MI control cohort using Cox proportional hazards and flexible parametric survival models., Findings: Among a total of 375,669 MI patients (130,758 females; 34.8%) and 1,878,345 matched non-MI patients (653,790 females; 34.8%), we identified 28,799 unique disease trajectories. The accrual of multiple circulatory diagnoses was more common amongst MI patients (RR 4.32, 95% CI 3.96-4.72) and conferred an increased risk of death (HR 1.32, 1.13-1.53) compared with matched controls. Trajectories featuring neuro-psychiatric diagnoses (including anxiety and depression) following circulatory disorders were markedly more common and had increased mortality post MI (HR ranging from 1.11 to 1.73) compared with non-MI individuals., Interpretation: These results provide an opportunity for early intervention targets for survivors of MI-such as increased focus on the psychological and behavioural pathways-to mitigate ongoing adverse disease trajectories, multimorbidity, and premature mortality., Funding: British Heart Foundation; Alan Turing Institute., Competing Interests: Declaration of interests CJH, JAB, DRW, OJ, JW, and MH have no conflicts of interest to declare. CPG has received funding from Abbott Diabetes, Bristol Myers Squibb and the European Society of Cardiology, and consulting fees from AI Nexus, AstraZeneca, Amgen, Bayer, Bristol Myers Squibb, Boehrinher-Ingleheim, CardioMatics, Chiesi, Daiichi Sankyo, GPRI Research B.V., Menarini, Novartis, iRhyth, Organon as well as payment for honoraria or lectures from AstraZeneca, Boston Scientific, Menarini, Novartis, Raisio Group, Wondr Medical, Zydus. CPG declares participation on Data Safety Monitoring or Advisory boards for the DANBLCOK and TARGET CTCA trials and editorial and committee membership of the NICE Indicator Advisory Committee, EHJ Quality of Care and Clinical Outcomes and ESC Quality Indicator Committee., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Neural systems are shaped by multiple constraints, balancing region communication with the cost of establishing and maintaining physical connections. It has been suggested that the lengths of neural projections be minimized, reducing their spatial and metabolic impact on the organism. However, long-range connections are prevalent in the connectomes across various species, and thus, rather than rewiring connections to reduce length, an alternative theory proposes that the brain minimizes total wiring length through a suitable positioning of regions, termed component placement optimization . Previous studies in nonhuman primates have refuted this idea by identifying a nonoptimal component placement, where a spatial rearrangement of brain regions in silico leads to a reduced total wiring length. Here, for the first time in humans, we test for component placement optimization. We show a nonoptimal component placement for all subjects in our sample from the Human Connectome Project ( N = 280; aged 22-30 years; 138 females), suggesting the presence of constraints-such as the reduction of processing steps between regions-that compete with the elevated spatial and metabolic costs. Additionally, by simulating communication between brain regions, we argue that this suboptimal component placement supports dynamics that benefit cognition., (© 2022 Massachusetts Institute of Technology.)

The increasing need for tissue substitutes in reconstructive surgery spurs the development of engineering methods suited for clinical applications. Cell culture and tissue production traditionally require the use of fetal bovine serum (FBS) which is associated with various complications especially from a translational perspective. Using the self-assembly approach of tissue engineering, we hypothesized that all important parameters of tissue reconstruction can be maintained in a production system devoid of FBS from cell extraction to tissue reconstruction. We studied two commercially available serum-free medium (SFM) and xenogen-free serum-free medium (XSFM) for their impact on tissue reconstruction using human adipose-derived stem/stromal cells (ASCs) in comparison to serum-containing medium. Both media allowed higher ASC proliferation rates in primary cultures over five passages compared with 10% FBS supplemented medium while maintaining high expression of mesenchymal cell markers. For both media, we evaluated extracellular matrix production and deposition necessary to engineer manipulatable tissues using the self-assembly approach. Tissues produced in SFM exhibited a significantly increased thickness (up to 6.8-fold) compared with XSFM and FBS-containing medium. A detailed characterization of tissues produced under SFM conditions showed a substantial 50% reduction of production time without compromising key tissue features such as thickness, mechanical resistance and pro-angiogenic secretory capacities (plasminogen activator inhibitor 1, hepatocyte growth factor, vascular endothelial growth factor, angiopoietin-1) when compared to tissues produced in the control FBS-containing medium. Furthermore, we compared ASCs to the frequently used human dermal fibroblasts (DFs) in the SFM culture system. ASC-derived tissues displayed a 2.4-fold increased thickness compared to their DFs counterparts. In summary, we developed all-natural human substitutes using a production system compatible with clinical requirements. Under culture conditions devoid of bovine serum, the resulting engineered tissues displayed similar and even superior structural and functional properties over the classic FBS-containing culture conditions with a considerable 50% shortening of production time., (Creative Commons Attribution license.)

Background: Skin tears are common in older adults and those taking steroids and warfarin. They are traumatic, often blunt injuries caused by oblique knocks to the extremities. The epidermis may separate from the dermis or both layers from underlying tissues leaving a skin flap or total loss of tissue, which is painful and prone to infection. 'Dermatuff™' knee-length socks containing Kevlar fibres (used in stab-proof vests and motorcyclists' clothing) aim to prevent skin tears. The acceptability of the socks and the feasibility of a randomised controlled trial (RCT) had not been explored., Methods: In this pilot parallel group RCT, 90 people at risk of skin-tear injury from Devon care homes and primary care were randomised to receive the socks or treatment as usual (TAU). The pilot aimed to estimate parameters to inform the design of a substantive trial and record professionals' views and participants' acceptability of the intervention and of study participation., Results: Participants were randomised from July 2013 and followed up until February 2015. Community participants were easier to recruit than care homes residents but were 10 years younger on average and more active. To recruit 90 participants, 395 had to be approached overall as 77% were excluded or declined. Seventy-nine participants (88%) completed the trial and 27/44 (61%) wore the socks for 16 weeks. There were 31 skin tear injuries affecting 18 (20%) of the 90 participants. The TAU group received more injuries, more repeated episodes, and larger tears with greater severity. Common daily diary reasons for not wearing the socks included perceived warmth in hot weather or not being available (holiday, in hospital, bed rest). Resource use data were obtainable and indicated that sock wearing gave a reduction in treatment costs whilst well-completed questionnaires showed improvements in secondary outcomes., Conclusions: This pilot trial has successfully informed the design and conduct of a future definitive cost-effectiveness RCT. It would need to be conducted in primary care with 880 active at-risk, elderly patients (440 per arm). Skin tear incidence and quality of life (from EQ5D5L) over a 4-month period would be the primary and secondary outcomes respectively., Trial Registration: ISRCTN, ISRCTN96565376.

Three metazoan parasites, a monogenean Hexostoma thynni and two species of copepods Pseudocycnus appendiculatus and Euryphorus brachypterus are known to parasitize the gills of ranched southern bluefin tuna (SBT) and other tuna species. However, there is no detailed information describing the pathological response to infection by these parasites in this species. Wild southern bluefin tuna Thunnus maccoyii (approximately 3 years of age), captured and towed to a grow-out site in the waters immediately south of Port Lincoln, South Australia were subsequently sampled ( n = 10) monthly from March until August 2004 during commercial harvest operations. Longitudinal sections of gill hemibranchs with attached parasites were excised and fixed for routine histology and immunohistochemistry. Reference samples were also collected from fish displaying no signs of parasitism or other grossly observable anomalies. Two morphologically distinct granulocytes were observed and putatively identified as eosinophils and mast cells. Pathology was localized to filaments upon and immediately adjacent to parasite attachment sites. Branchial cellular responses, adjunct to the attachment of H. thynni by its opisthaptoral clamps, included hyperplasia and inflammation resulting in structural remodeling of branchial tissues. Inflammatory infiltrates were often dominated by putative eosinophils and lymphocytes when parasitized by H. thynni and P. appendiculatus . Gill associated lymphoid tissue infiltrated the lamellar regions particularly in response to helminth infection. A variable response ranging from hemorrhage with minor hyperplasia or fibroplasia and eosinophilic inflammation to a barely discernible change was seen for gill sections harboring P. appendiculatus and E. brachypterus . The magnitude of the host response to attachment by the latter was congruent with attachment proximity and parasite load. On the basis of the host responses reported here and the low intensity of infection observed in other associated studies these gill ectoparasites are currently considered a low risk for wild and ranched adult SBT.

Three species of Pseudodactylogyrus Gusev, 1965 (Monogenea: Pseudodactylogyridae) were collected from the gills of Anguilla reinhardtii Steindachner and A. australis Richardson from several localities in Australia and eels imported to Japan from Australia. Pseudodactylogyrus gusevi sp. n. from A. reinhardtii (type host) and A. australis in Queensland, Australia is most similar to P. bini (Kikuchi, 1929), but can be differentiated by the shorter male copulatory tube, heavy sclerotisation of the vaginal tube and the presence of a small projection of the supplementary piece of the hamulus. Pseudodactylogyrus rohdei sp. n. from A. australis (type host) in Queensland, Australia is most similar to P. anguillae (Yin et Sproston, 1948), but differs in the possession of a longer cement gland and the presence of a small projection on the supplementary piece of the hamulus. Pseudodactylogyrus bini sensu Gusev, 1965 and P. anguillae sensu Gusev, 1965 are synonymised with P. gusevi sp. n. and P. rohdei sp. n., respectively. Pseudodactylogyrus mundayi sp. n. from A. australis, originating in Tasmania, Australia and sent alive to Japan, is most similar to P. kamegaii Iwashita, Hirata et Ogawa, 2002, from which it can be discriminated by the shorter male copulatory tube and the shorter vaginal tube. Dactylogyrus bialatus Wu, Wang et Jian, 1988 from Synechogobius ommaturus (Richardson) (Gobiidae) is transferred to Pseudodactylogyrus as P. bialatus comb. n. A phylogenetic tree based on the ITS2 region of six species of Pseudodactylogyrus including P. gusevi and P. mundayi shows that P. haze from a goby diverged first, and that species from eels are monophyletic, forming three lineages differing by their zoogeographical distribution. With the three new species and one new combination proposed in this paper, Pseudodactylogyrus is now comprised of eight species infecting anguillid and gobiid fish, and a key to species is presented.

A survey of blood fluke, Cardicola forsteri, infection in ranched southern bluefin tuna, Thunnus maccoyii, was undertaken over three farming seasons, from March 2004 to September 2006. Analyses of covariance and logistic regression were used to explore the effects of company, year, season, time in culture, and condition index on intensity, abundance and prevalence of blood fluke infection. Average prevalence of blood fluke infection was 62.64% over the period of the survey. Average intensity was 6.20 (± 0.57) fluke per infected host and the average abundance was 3.70 (± 0.57) fluke per host. Year did not influence mean intensity or abundance although a significant decrease in prevalence in 2005 was evident. Tuna harvested in winter had a significantly greater abundance and prevalence of blood fluke than the tuna harvested in autumn. No effect of intensity or abundance of infection was observed on the condition of the infected tuna. A universal factor in explaining variation in C. forsteri intensity, abundance and prevalence was company. Differences in infection levels between tuna from different companies may be related to differences in husbandry measures employed on each farm, or due to different average sizes of tuna farmed by each of the companies, or due to the location of the operations., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Skin tears are traumatic injuries occurring mostly on the extremities due to shearing and friction forces that separate the epidermis and the dermis from underlying tissues. They are common and occur mostly in older adults and those taking medications that compromise skin integrity. Pretibial skin tears can develop into leg ulcers, which require lengthy, expensive treatment to heal. Traumatic injuries are the second most common type of wounds after pressure ulcers in care homes and are the commonest reason for older adults to require the attention of a community nurse. Common causes of skin tear injuries are bumping into furniture and other obstacles, using mobility aids, transfer to/from wheelchairs, getting in and out of bed and falls. No effective preventative measures currently exist but knee-length, protective socks are now available that contain impact-resistant Kevlar fibres (of the type used in stab-proof vests) and cushioning layers underneath., Methods/design: In this pilot parallel group, randomised controlled trial, 90 people at risk of skin-tear injury will be randomised with equal allocation to receive the intervention or usual care. They will be recruited from care homes and from the community via general practices and a research volunteer database. Pilot outcomes include recruitment, eligibility, attrition, ascertainment of injuries and completion of outcome measures. Acceptability of the intervention and of study participation will be explored using semi-structured interviews. The proposed primary outcome for the future definitive trial is skin tear-free days. Secondary outcomes are skin tear severity, health status, specific skin-tears quality of life, capability and fear of falling, measured at baseline and the end of the study and in the event of a skin tear., Discussion: The results of this study will be used to inform the development and design of a future randomised controlled trial to assess the effectiveness and cost-effectiveness of a unique and innovative approach to skin tear prevention. Approval was granted by the NRES - Cornwall and Plymouth Research Ethics Committee (13/SW/013). Dissemination will include publication of quantitative and qualitative findings, and experience of public involvement in peer-reviewed journals., Trial Registration: Current Controlled Trials: ISRCTN96565376.

The epithelial cells and Wharton׳s jelly cells (WJC) from the human umbilical cord have yet to be extensively studied in respect to their capacity to generate tissue-engineered substitutes for clinical applications. Our reconstruction strategy, based on the self-assembly approach of tissue engineering, allows the production of various types of living human tissues such as skin and cornea from a wide range of cell types originating from post-natal tissue sources. Here we placed epithelial cells and WJC from the umbilical cord in the context of a reconstructed skin substitute in combination with skin keratinocytes and fibroblasts. We compared the ability of the epithelial cells from both sources to generate a stratified, differentiated skin-like epithelium upon exposure to air when cultured on the two stromal cell types. Conversely, the ability of the WJC to behave as dermal fibroblasts, producing extracellular matrix and supporting the formation of a differentiated epithelium for both types of epithelial cells, was also investigated. Of the four types of constructs produced, the combination of WJC and keratinocytes was the most similar to skin engineered from dermal fibroblasts and keratinocytes. When cultured on dermal fibroblasts, the cord epithelial cells were able to differentiate in vitro into a stratified multilayered epithelium expressing molecules characteristic of keratinocyte differentiation after exposure to air, and maintaining the expression of keratins K18 and K19, typical of the umbilical cord epithelium. WJC were able to support the growth and differentiation of keratinocytes, especially at the early stages of air-liquid culture. In contrast, cord epithelial cells cultured on WJC did not form a differentiated epidermis when exposed to air. These results support the premise that the tissue from which cells originate can largely affect the properties and homoeostasis of reconstructed substitutes featuring both epithelial and stromal compartments., (Copyright © 2014 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)

Introduction: Pain is the commonest reason that patients present to an emergency department (ED), but it is often not treated effectively. Patient controlled analgesia (PCA) is used in other hospital settings but there is little evidence to support its use in emergency patients. We describe two randomised trials aiming to compare PCA to nurse titrated analgesia (routine care) in adult patients who present to the ED requiring intravenous opioid analgesia for the treatment of moderate to severe pain and are subsequently admitted to hospital., Methods and Analysis: Two prospective multi-centre open-label randomised trials of PCA versus routine care in emergency department patients who require intravenous opioid analgesia followed by admission to hospital; one trial involving patients with traumatic musculoskeletal injuries and the second involving patients with non-traumatic abdominal pain. In each trial, 200 participants will be randomised to receive either routine care or PCA, and followed for the first 12 h of their hospital stay. The primary outcome measure is hourly pain score recorded by the participant using a visual analogue scale (VAS) over the 12 h study period, with the primary statistical analyses based on the area under the curve of these pain scores. Secondary outcomes include total opioid use, side effects, time spent asleep, patient satisfaction, length of hospital stay and incremental cost effectiveness ratio., Ethics and Dissemination: The study is approved by the South Central-Southampton A Research Ethics Committee (REC reference 11/SC/0151). Data collection will be completed by August 2013, with statistical analyses starting after all final data queries are resolved. Dissemination plans include presentations at local, national and international scientific meetings held by relevant Colleges and societies. Publications should be ready for submission during 2014. A lay summary of the results will be available to study participants on request, and disseminated via a publically accessible website., Registration Details: The study is registered with the European Clinical Trials Database (EudraCT Number: 2011-000194-31) and is on the ISCRTN register (ISRCTN25343280).

The human umbilical cord (UC) has attracted interest as a source of cells for many research applications. UC solid tissues contain four cell types: epithelial, stromal, smooth muscle and endothelial cells. We have developed a unique protocol for the sequential extraction of all four cell types from a single UC, allowing tissue reconstruction using multiple cell types from the same source. By combining perfusion, immersion and explant techniques, all four cell types have been successfully expanded in monolayer cultures. We have also characterised epithelial and Wharton's jelly cells (WJC) by immunolabelling of specific proteins. Epithelial cell yields averaged at 2.3 × 10(5) cells per centimetre UC, and the cells expressed an unusual combination of keratins typical of simple, mucous and stratified epithelia. Stromal cells in the Wharton's jelly expressed desmin, α-smooth muscle actin, elastin, keratins (K12, K16, K18 and K19), vimentin and collagens. Expression patterns in cultured cells resembled those found in situ except for basement membrane components and type III collagen. These stromal cells featured a sustained proliferation rate up to passage 12 after thawing. The mesenchymal stem cell (MSC) character of the WJC was confirmed by their expression of typical MSC surface markers and by adipogenic and osteogenic differentiation assays. To emphasise and demonstrate their potential for regenerative medicine, UC cell types were successfully used to produce human tissue-engineered constructs. Both bilayered stromal/epithelial and vascular substitutes were produced, establishing the versatility and importance of these cells for research and therapeutic applications., (Copyright © 2012 S. Karger AG, Basel.)

The blood fluke, Cardicola forsteri, is a prevalent infection in ranched southern bluefin tuna. This project aimed to define the timing and intensity of the various developmental stages of C. forsteri within southern bluefin tuna as well as to relate infection to host pathology and immune response. Archival samples from several cohorts of T. maccoyii sampled from 2008 to 2010 were used in this study. The prevalence and intensity of C. forsteri infection was described using heart flushes and histological examination. Humoral immune response, i.e. C. forsteri specific antibody, lysozyme activity, and alternative complement activity, was also described. Based on the validated and detailed C. forsteri infection timeline, relationships between infection events, physiological response, and diagnosis were proposed. Immune response developed concurrently with C. forsteri infection, with the majority of physiological response coinciding with commencing egg production. Further research is needed to confirm the origin of C. forsteri antigen which is responsible for immune response development and how T. maccoyii immune response works against infection. To aide this research, further diagnostic methods for confirmation of infection need to be developed.

Ranched southern bluefin tuna Thunnus maccoyii were fed baitfishes supplemented with vitamins (predominantly E and C) or vitamins and immunostimulants, nucleotides and β-glucans, over 12 weeks after transfer and monitored for enhancement in immune response, health and performance through their 19 week grow-out period. Fish from two different tows were sampled separately at three different sampling points: at transfer to grow-out pontoons, at 8 weeks post-transfer and at harvest, 19 weeks post-transfer. Lysozyme activity was enhanced during vitamin supplementation compared to control fish. Performance (i.e. survival, condition index and crude fat), health (i.e. blood plasma variables including pH, osmolality, cortisol, lactate and glucose) and alternative complement activity were not commonly improved through diet supplementation. There were some tow-specific improvements in performance through vitamin supplementation including survival, selected parasite prevalence and intensity, and alternative complement activity. Immunostimulant supplementation also showed a tow-specific improvement in plasma cortisol level. Tow-specific responses may suggest that life history, previous health condition and husbandry can affect the success of vitamin and immunostimulant enhancement of immune response, health and performance of ranched T. maccoyii., (© 2011 The Authors. Journal of Fish Biology © 2011 The Fisheries Society of the British Isles.)

Aporocotylids (fish blood flukes) are emerging as pathogens of fishes in both marine and freshwater aquaculture. Efforts to control these parasites are hampered by a lack of life cycle information. Here we report on the life cycle of Cardicola forsteri, which is considered a significant pathogen in southern bluefin tuna, Thunnus maccoyi, ranched in South Australia. We surveyed polychaetes, bivalves and gastropods from sites close to tuna pontoons. Infections consistent with the Aporocotylidae were found in terebellid polychaetes, a single Longicarpus modestus and five individuals of Reterebella aloba. All infections were comprised of hundreds of sporocysts in the body cavity of the host, each filled with developing and mature cercariae. Sequences of ITS-2 and lsrDNA from the infection from L. modestus were a perfect match with those of adult C. forsteri from T. maccoyi. This life cycle link is considered confirmed but it is possible that additional terebellid species are infected in South Australia; equally, other species of intermediate host are likely to be involved in other parts of the range of this cosmopolitan trematode. Sequences of the species from R. aloba did not match a known adult but phylogenetic analysis of lsrDNA suggests that it is also a species of Cardicola Short, 1953. These findings show that terebellid polychaetes are a major host group for marine aporocotylids, especially given that Cardicola is the largest marine aporocotylid genus. The two cercarial types are among the smallest known for the family and are unusual, but not unique, in having short, simple tails. We speculate that the form of the tail means that these cercariae are not active swimmers and are thus heavily dependent on currents for dispersal. Control of this parasite might be effected by moving the tuna pontoons appropriate distances to avoid encounter with current-dispersed cercariae, or by increasing the separation of the nets from the sea floor, either by raising the nets or moving to deeper water., (Copyright © 2011 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

Peaks in epizootics of sea lice (mostly Caligus chiastos) and blood flukes (Cardicola forsteri) among Southern Bluefin tuna (Thunnus maccoyii) appear to coincide with the onset of an increased mortality. The mortality event occurs 6-12 weeks after T. maccoyii have been transferred into static ranching pontoons from the wild. However, to date available data on parasite occurrence before commercial harvesting begins, are scant. This research gathered epizootiological data from weeks 4 to 13 post-transfer, for 153 T. maccoyii sampled from two research and four commercial pontoons. Counts of both parasites in the research pontoons reached levels far heavier than previously documented in ranched T. maccoyii. For sea lice, the prevalence in most pontoons was 100%; the highest intensity reached 495 individuals, and mean counts at the peak of the infection exceeded 265 lice per fish. Almost all of the 5407 individual lice counted were identified as adult C. chiastos (89.44% female, 10.14% male); adult females of two other species were also present, C. amblygenitalis (0.13%), in addition to an undescribed species, C. sp. (0.04%). Lice counts were correlated positively with gross eye pathology scores (r(s,151df)=0.3394, p=0.0000), negatively correlated with condition index (r(s,151df)=-0.5396, p=0.0000), and positively correlated with plasma cortisol (r(s,131df)=0.3906, p=0.0000) and glucose (r(s,131df)=0.2240, p=0.0096). For the blood fluke, prevalences were less uniform than those of sea lice, with lower rates of infection at the beginning (ranging from 10% to 40%), reaching 100% mid-study, and declining again (40% in one pontoon). The highest intensity reached 441 individual flukes. Fluke counts were negatively correlated with plasma haemoglobin (r(s,151df)=-0.2436, p=0.0051) and positively with lysozyme (r(s,151df)=0.3013, p=0.0019). Fluke counts were also correlated with sea lice counts (r(s,150df)=0.3143, p=0.0000). Peaks in these epizootics occurred near the onset of elevated mortalities, which started after 7 weeks of ranching., (Copyright 2010 Elsevier B.V. All rights reserved.)

The presently reported study provides a detailed morphological description of the female and the male of a new species of the genus Parabrachiella-Parabrachiella jarai sp. nov. The parasites were sampled from marine fish, silver sillago, Sillago sihama (Perciformes: Sillaginidae), captured in Malaysia in 1994 and Hong Kong in 1995. The new species bears some resemblance to Parabrachiella lata (Song et Chen, 1976) but differs from it in details of second antenna, mandible, and maxilliped. The genus Parabrachiella currently covers 67 species including those recently transferred from Neobrachiella Kabata, 1979. An amended generic diagnosis is proposed for Parabrachiella and Thysanote. Some members of Parabrachiella are herewith transferred to Thysanote and some Thysanote are now placed in Parabrachiella., (Copyright 2009 Elsevier Inc. All rights reserved.)

Within the typical 2-8 month (January to August inclusive) farming cycle for southern bluefin tuna, Thunnus maccoyii (Castelnau), in Spencer Gulf, South Australia, counts of a sea louse, Caligus chiastos Lin et Ho, 2003, were strongly statistically associated with both fish condition and severity of eye damage. During a trial examining the feasibility of maintaining T. maccoyii in farms for more than 1 year, including over the summer season when temperatures may exceed 24 degrees C, we collected additional epidemiological data on burdens of sea lice over a 17-month period (April 2005 to August 2006 inclusive), on a total of 200 T. maccoyii and 40 'control'T. maccoyii farmed and harvested within 2006. In the first farming season, an epizootic of C. chiastos was characterized by a significant increase in prevalence from 0% to 55% in the first 6 weeks after transfer to farms from the wild, which was followed by a significant decline to zero over the next 12 weeks. A single specimen of a second species of Caligus was also detected within this 4.5-month period. In the second farming season, we recorded a third species of sea louse, C. amblygenitalis Tripathi, 1961. In March 2006, a second epizootic peak occurred, this time with mixed infections of C. chiastos and C. amblygenitalis, with a combined prevalence of 100%. The prevalence of both sea lice species then declined significantly over the second winter period (June to August inclusive). On all but one date that sea lice were detected, sea lice counts were significantly associated with the severity of gross eye damage. Because both peaks in infection occurred in summer months (December to February inclusive), we conclude that infections of sea lice pose a risk to the farming of T. maccoyii under certain summer conditions within Spencer Gulf.

In this study, adaptive immune response was investigated in farmed southern bluefin tuna, Thunnus maccoyii, infected with a sanguinicolid Cardicola forsteri. A cohort (Cohort(2005)) of southern bluefin tuna was sampled between March 2005 and August 2006. Samples were taken at the transfer of wild caught tuna to sea cages and then at regular intervals. Parasite intensity, abundance and prevalence data were recorded. An ELISA was developed to detect and quantify an antibody response against the blood fluke in southern bluefin tuna serum. Intensity and prevalence of the blood fluke were shown to peak in May 2005 at 10.9 flukes per infected fish (SE=1.72) and 97.5% prevalence and then decreased to low prevalence (10%) and intensity (1.0). There were no significant changes in prevalence or intensity in 2006. Antibody titres and seroprevalence increased from 1.37 U microl(-1) and 10% at transfer in March 2005 to reach a peak in December 2005 of 25.86 U microl(-1) (SE=6.26 U microl(-1)) and 66.66%. No significant changes were observed in antibody titres for the same cohort of fish during 2006. Parasitological and serological values from Cohort(2005) were compared to a 2006 cohort (Cohort(2006)) in March 2006 and August 2006 to determine if prior infection in Cohort(2005) elicited any protection against infection in 2006. Although significant differences were not observed in intensities between cohorts it was shown that Cohort(2005) had significantly lower abundances and prevalences of blood fluke infection than Cohort(2006). Although there was no significant difference in mean antibody titres between cohorts in March 2006, the mean antibody titre of Cohort(2006) was significantly greater than that of Cohort(2005) in August 2006. No significant differences were observed in seroprevalence. This is one of the few studies to demonstrate the development of acquired resistance in fish against a parasite in an aquaculture environment under natural infection conditions.

Tuna farming off Port Lincoln, Australia, involves catching wild 2-4-year-old southern bluefin tuna in summer and then fattening for periods of 2-8 months. As fresh product is not available year-round, the feasibility of maintaining tuna for longer periods was trialled, including over a summer season, when temperatures may exceed 24 degrees C. As the rates of growth and reproduction in ectoparasites of fishes are usually most rapid during warm temperatures, parasite epizootics at this time may adversely affect the health of tuna. We collected epidemiological data on burdens of metazoans on the gills of tuna from the time of stocking in April 2005 through to final harvest in August 2006 (N=220). We document an epizootic of the copepod Pseudocycnus appendiculatus, characterised by a significant increase in both prevalence and mean intensity in the first winter, followed by a decline in these parameters over the next 12 months. This epizootic pattern appears to be independent of seasonal changes in temperature. For two other species, a second copepod (Euryphorus brachypterus) and a polyopisthocotylean flatworm (Hexostoma thynni), there were no clearly discernible trends in infections. As the high water temperatures over the summer period did not lead to increased infections of any species of gill parasites, we conclude that they do not threaten the feasibility of farming of Thunnus maccoyii.

A new preparation is described for recording the electroretinogram (ERG) from larval zebrafish (5-8 days postfertilization) which has allowed the investigation of the pharmacology of cone photoreceptor inputs onto bipolar cells. By using a pharmacological cocktail to isolate the photoreceptors and bipolar cells from inhibitory influences, it was found that an excitatory amino acid transporter (EAAT) presumably linked to a Cl() channel mediates most of the synaptic transmission from the cone photoreceptors to the ON bipolar cells, although metabotropic glutamate receptors (presumably mGluR6) also make a small contribution. On the other hand, alpha-amino-3-hydroxy- 5-methyl-4-isoxazolepropionate (AMPA)/kainate receptors mediate synaptic transmission from cone photoreceptors to OFF bipolar cells. The glutamatergic input mechanisms underlying bipolar cell responses in the larval zebrafish are adultlike and similar to those in other teleost species.

Much is known about those aspects of tuna health which can be studied in wild populations, e.g. helminth parasites. However, because aquaculture of these species is in its infancy, knowledge of microbial, nutritional and environmental diseases is limited. This review is an attempt to bring together the available information on those diseases of Thunnus spp. which cause significant morbidity, mortality or economic loss. In doing so it has become clear that much more research needs to be undertaken on the physiology of the species (southern, northern and Pacific bluefin tuna) currently used in aquaculture in order for the pathogenesis of some conditions to be properly understood. Attempts at hatchery culture of Pacific bluefin tuna has indicated that Thunnus spp. will be problematic to hatch and propagate.

Lernanthropus polynemi Richiardi, 1881 is described, for the first time since its original discovery, based on the type-material found in the Museum für Naturkunde in Berlin, Germany. Morphological features of this parasite, which infects the commercially-caught fish Eleutheronema tetradactylum (Shaw), are described and illustrated, including those of the male, which is described for the first time. The types of another species from the same host, L. lappaceus Wilson, 1912, and deposited in the National Museum of Natural History in Washington DC, are also examined based on the type-specimens. The types of a third congener, L. trifoliatus Basset-Smith, 1898, also from the same host, are not available, but the original illustrations are studied. It is considered that the three nominal species described from E. tetradactylum represent a single taxon, and that L. lappaceus and L. trifoliatus be relegated to synonymy with L. polynemi.

Four genera of scombrid fishes (26 species) and their copepod (32 species) and monogenean ectoparasites (25 species) were used to test the hypothesis that the East Pacific Barrier is responsible for the most pronounced break in the circum-tropical warm water fauna of the continental shelves, and not the New World Barrier. Analysis at the species level showed that there is a primary centre of diversity in the West Pacific, and a secondary centre in the West Atlantic. The former, almost entirely, shares its species of the largely coastal Scomberomorus and Grammatorcynus and their parasites with seas located to the west. Only four parasites (all copepods) are shared by the East and West Pacific, and they are circum-tropical. In contrast, the West Pacific shares species of the more pelagic Scomber and their parasites with seas both to the east and west, although at the genus level, only two circum-tropical monogenean genera are shared by the E and W Pacific. We conclude that the East Pacific Barrier has been a 100% or almost 100% effective barrier to dispersal of species of Scomberomorus, Grammatorcynus and their parasites, whereas for species of Scomber and their parasites, the East Pacific has been a less effective barrier.

Members of the subfamily Priceinae are gastrocotylinean monogeneans of the gills of scombrid fishes of the genus Scomberomorus (and perhaps the genera Acanthocybium, Rastrelliger and Katsuwonus) from warm to warm-temperate seas of the world. We revise the diagnosis of the subfamily and regard the Mexicotylinae Lebedev, 1984 as a synonym. Two monotypic genera are accepted as valid. Pricea multae Chauhan, 1945 is recorded from seven species of Scomberomorus from the Indo-west Pacific, from off eastern South Africa north to the Persian Gulf and as far east as Fiji. New synonyms we recognise include P. minimae Chauhan, 1945 (described from India, reportedly on Katsuwonus pelamis), P. solandri Gupta & Chanana, 1977 (a single specimen was described from India, reportedly on Acanthocybium solandri) and P. microcotylae Chauhan, 1945 (also described from India, reportedly on Rastrelliger kanagurta). Mexicotyle mexicana (Meserve, 1938) Lebedev, 1984 is recorded on four species of Scomberomorus from the western Atlantic Ocean (United States to Brazil), two in the eastern Pacific (California to Peru) and one from the eastern Atlantic (Ghana).

Members of the subfamily Thoracocotylinae are gastrocotylid monogeneans of Spanish mackerels (scombrid fishes of the genus Scomberomorus) from warm to warm-temperate seas around the world. We revise the diagnosis of the subfamily and recognise two genera and three species as valid. The genus Paradawesia Bravo Hollis & Lamothe Argumedo, 1976 is synonymised with Thoracocotyle MacCallum, 1913, and Dawesia Unnithan, 1965 and Methoracocotyle Lebedev, 1984 are synonymised with Pseudothoracocotyla Yamaguti, 1963. Thoracocotyle crocea MacCallum, 1913 (syns T. paradoxica Meserve, 1938 and Paradawesia bychowskyi Bravo Hollis & Lamothe Argumedo, 1976) is recorded from two species of Scomberomorus in the eastern Pacific (California to Peru) and four in the western Atlantic (South Carolina to Brazil). Pseudothoracocotyla ovalis (Tripathi, 1956) Yamaguti, 1963 (new syns Dawesia indica Unnithan, 1965, D. incisa Lebedev, 1970, Methoracocotyle scomberomori (Young, 1968) Lebedev, 1984, M. gigantica (Rohde, 1976) Lebedev, 1984 and Thoracotyle indica (Unnithan, 1965) Murugesh, 1995) is recorded from the gills of seven species of Scomberomorus from the Indo-west Pacific (eastern South Africa north to the Persian Gulf, and east to Fiji). Pseudothoracocotyla whittingtoni n. sp. is described from an eighth Indo-west Pacific scomberomorid, S. munroi, in Australian waters.

Members of the subfamily Neothoracocotylinae are gastrocotylinean monogeneans on the gills of scombrid fishes of the genera Scomberomorus and Acanthocybium, and reportedly of a coryphaenid fish belonging to the genus Coryphaena. We revise the diagnosis of the subfamily and its two genera and accept only two species as valid. Neothoracocotyle acanthocybii (Meserve, 1938) Hargis, 1956 is known from Acanthocybium solandri throughout the Pacific Ocean and in the western Atlantic. N. coryphaenae (Yamaguti, 1938) Hargis, 1956, known only from a single specimen and described from Coryphaena hippurus in Japan, is synonymised with N. acanthocybii. The sole member of Scomberocotyle, S. scomberomori (Koratha, 1955) Hargis, 1956, infects five species of Scomberomorus in the eastern Pacific Ocean and the western and castern Atlantic. We record this worm from several new hosts and/or localities, including S. sierra and S. concolor in the eastern Pacific (Mexico to Colombia), S. maculatus and S. cavalla in the western Atlantic (USA to Brazil), and S. tritor in the eastern Atlantic (Sierra Leone to Nigeria).

Scomberomorocotyle munroi n. g., n. sp. is described from the gills of Scomberomorus munroi, a Spanish mackerel from the coasts of northern Australia and southern Papua New Guinea. The genus belongs to the suborder Gastrocotylinea because a pair of basal accessory sclerites is present in the clamps. However, the worm does not belong to any of the eight gastrocotylinean families as they are currently recognised. The worm appears to be a member of the Thoracocotylidae, in that the male copulatory organ has relatively weakly developed spines, and that the haptor is one-sided with two rows of clamps. However, the worm differs from all thoracocotylids in that the clamps lack the characteristic lateral rib-like thickenings. To accommodate the new genus and species, the diagnosis of the Thoracocotylidae Price is amended to include worms lacking ribs in their clamps, and a new subfamily, the Scomberomorocotylinae n. subfam., is erected; a key to the four subfamilies which we recognise as valid is provided.

Four-hundred and fifty-three Scomber australasicus, ranging in length from 14.1 to 46.5 cm and taken in 12 samples over 5 years from a single locality in south-eastern Australia, were examined. Ten species of ectoparasites were recorded: six monogeneans, three copepods and one isopod. The maximum number of parasite species in any one fish reached five, and the maximum total parasite intensity reached 39. Host size is an important determinant of the structure of ectoparasite assemblages of slimy mackerel: the average number of species per host peaked at 2.2 in fishes between 20 and 25 cm in length, then declined in larger fish; the abundance of all parasites on each fish similarly peaked in fishes 20-25 cm (mean of 9.9). The monogenean Pseudokuhnia minor had the highest prevalence and abundance of all parasites, infecting almost 80% of fish < 25 cm long. Over half of the total number of parasites belonged to this species, and it was dominant in intensity in just over half of the fish in which it occurred. When approximate volume was considered, assemblages were dominated by Kuhnia scombercolias in fish < 20 cm, by Kuhnia scombri in fish 20-34.9 cm, and by P. minor in fish larger than 35 cm long. The intensities of these three monogeneans were very strongly and positively correlated with each other, as were the intensities of P. minor and Grubea australis; the intensities of only one pair of species were associated negatively (K. scombercolias and the isopod Ceratothoa imbricata). A nested subset analysis indicates that the ectoparasite assemblages are random. This and the low prevalences and abundances of infection, as well as low species richness per fish, indicate that infra-assemblages are isolationist.

Just nineteen species of ectoparasitic helminths were found in a survey of over 1,500 individuals of 26 species of sillaginid fishes in the Indo-west Pacific. A twentieth worm is known only from the literature; a twenty-first, also known only from the literature, is considered a doubtful record. Fifteen of the twenty worms are branchial monogeneans, one is a monogenean of the pharyngeal plates, one is an ectoparasitic digenean living under the scales, and three are leeches of the mouth cavity and fins. The most common monogeneans were diplectanids (Diplectanum spp. and Monoplectanum spp.) and microcotylids (Polylabris spp.), each with five recently described or redescribed species. Of the remaining monogeneans, three were extremely rare, and two were uncommon. Pseudobivagina sp. and Polynemicola sp. (Microcotylidae) and Pseudempleurosoma sp. (Ancyrocephalidae) were represented by only a single worm each from three different hosts (Sillago robusta, S. sihama, and S. ingenuua, respectively). The gyrodactylid Gyrodactylus sp. is widespread and was recorded from four species of sillaginids (S. ciliata, S. japonica, S. schomburgkii and S. sihama). Encotyllabe chironemi Robinson (Capsalidae) is recorded for the first time from sillaginids, but only on S. aeolus. Two additional monogeneans are known from sillaginids only in the literature: Dactylogyrus sp. (Dactylogyridae) is known only from cultured S. sihama; the single specimen of Microcotyle sp. (Microcotylidae) recorded from Sillaginodes punctata is probably a contaminant, since the haptor was missing. The generalist trematode Transversotrema licinum Manter (Transversotrematidae) was found for the first time in samples of four species of sillaginids (Sillago analis, S. ingenuua, S. lutea and S. sihama). Three species of piscicolid leeches were encountered: Austrobdella translucens Badham was common on the fins of three large inshore sillaginids (S. ciliata, S. schomburgkii and S. analis); a single specimen of the generalist species Zeylanicobdella arugamensis De Silva was recovered from S. soringa; and specimens of Z. stellata (Moore) infected S. schomburgkii and S. analis. The diversity of host-specific worms in Sillaginidae is low compared with those of some other Indo-west Pacific fishes.

The pharmacokinetics and tolerability of 1-g doses of ceftriaxone diluted in sterile water, 1% lidocaine, or buffered lidocaine were investigated. No difference in bioequivalence was noted between the three treatments. No difference in peak creatine kinase values was seen. By use of a quantitative pain scale, injection of ceftriaxone with the water diluent was significantly more painful than that with either of the other two diluents. No difference in injection pain was noted for lidocaine or buffered lidocaine.

Three monkey kidney cell lines, Vero, GL-V3 and MA-104 were subjected to karyological analysis to determine their chromosomal stability and to confirm their species of origin. Although the lines were shown to be relatively stable throughout all of the passage levels that were tested, the species of origin of one of them was found to be different from that claimed by the originators. This finding was supported by data from isoenzyme studies.