Writing for publication: Argument and evidence.

The rules for writing a research report are well defined but novice writers need more clarification on writing scholarly scientific papers for publication. The purpose of this paper is to enable novice writers to confidently apply the skills of scientific writing to the development of a scholarly paper for publication. We highlight the importance of having a clearly defined question or clarity in focus before moving on to consider the components of a scholarly paper including, the question, thesis, introduction, body, discussion, conclusion and finally, an abstract.

Sense of coherence moderates job demand-resources and impact on burnout among nurses and midwives in the context of the COVID-19 pandemic: A cross-sectional survey.

AIM: This study aimed to test the propositions using the job demands-resources (JD-R) model for main/moderation/mediation effects of a sense of coherence and practice environment support on mental well-being (anxiety, depression and burnout) outcomes in nurses and midwives in Australia during the COVID-19 pandemic. DESIGN: Cross-sectional quantitative survey. DATA SOURCES: The study was a cross-sectional design using self-report questionnaires reported as per the Reporting of Observational Studies in Epidemiology Guidelines. Following human research ethics approval (2020.ETH.00121) participants were recruited to take part in an online anonymous survey using self-report instruments to test the JD-R model in Australia. RESULTS: 156 participant nurses and midwives experienced anxiety, depression and emotional burnout during COVID-19. While a considerable proportion of participants indicated high levels of emotional exhaustion, their responses showed low levels of depersonalization (detached response to other people) and high levels of personal accomplishment (high levels of work performance and competence). A sense of coherence was a significant protective factor for mental health well-being for the participants, which is to say, high levels of sense of coherence were predictive of lower levels of anxiety, depression and burnout in this study sample. CONCLUSION: It is evident that both nursing and midwifery professions require psychosocial support to preserve their health both in the short and long term. Ensuring individualized tailored support will require a layered response within organizations aimed at individual self-care and collegial peer support. PATIENT OR PUBLIC CONTRIBUTION: There was no patient or public contribution in this study, as the focus was on nurses and midwives.

Turnovers of Sex-Determining Mutation in the Golden Pompano and Related Species Provide Insights into Microevolution of Undifferentiated Sex Chromosome.

The suppression of recombination is considered a hallmark of sex chromosome evolution. However, previous research has identified undifferentiated sex chromosomes and sex determination by single SNP in the greater amberjack (Seriola dumerili). We observed the same phenomena in the golden pompano (Trachinotus ovatus) of the same family Carangidae and discovered a different sex-determining SNP within the same gene Hsd17b1. We propose an evolutionary model elucidating the turnover of sex-determining mutations by highlighting the contrasting dynamics between purifying selection, responsible for maintaining W-linked Hsd17b1, and neutral evolution, which drives Z-linked Hsd17b1. Additionally, sporadic loss-of-function mutations in W-linked Hsd17b1 contribute to the conversion of W chromosomes into Z chromosomes. This model was directly supported by simulations, closely related species, and indirectly by zebrafish mutants. These findings shed new light on the early stages of sex chromosome evolution.

Optimising physiotherapy for people with lateral elbow tendinopathy - Results of a mixed-methods pilot and feasibility randomised controlled trial (OPTimisE).

BACKGROUND: The OPTimisE intervention was developed to address uncertainty regarding the most effective physiotherapy treatment strategy for people with Lateral Elbow Tendinopathy (LET). OBJECTIVES: To assess the feasibility of conducting a fully-powered randomised controlled trial (RCT) evaluating whether the OPTimisE intervention is superior to usual physiotherapy treatment for adults with LET. DESIGN: A mixed-methods multi-centred, parallel pilot and feasibility RCT, conducted in three outpatient physiotherapy departments in the UK. METHOD: Patients were independently randomised 1:1 in mixed blocks, stratified by site, to the OPTimisE intervention or usual care. Outcomes were assessed using pre-defined feasibility progression criteria. RESULTS: 50 patients were randomised (22 Female, 28 Male), mean age 48 years (range 27-75). Consent rate was 71% (50/70), fidelity to intervention 89% (16/18), attendance rate in the OPTimisE group 82% (55/67) vs 85% (56/66) in usual care, outcome measure completion 81% (39/48) at six-month follow-up. There were no related adverse events. Patients and physiotherapists reported that the OPTimisE intervention was acceptable but suggested improvements to the trial design. 49 patients were recruited from physiotherapy referrals vs one from primary care records. Outcome measure return rates were higher when completed online (74%) compared to postal questionnaire (50%). Exploratory analysis showed improvements in both groups over time. CONCLUSIONS: It is methodologically feasible to conduct a fully powered RCT comparing the clinical and cost-effectiveness of the OPTimisE intervention versus usual physiotherapy treatment. Considering the similar improvements observed in both groups, careful consideration is needed regarding the priority research question to be addressed in future research.

Probiotics enhance resistance to Streptococcus iniae in Nile tilapia (Oreochromis niloticus) reared in biofloc systems.

Biofloc technology is a rearing technique that maintains desired water quality by manipulating carbon and nitrogen and their inherent mixture of organic matter and microbes. Beneficial microorganisms in biofloc systems produce bioactive metabolites that may deter the growth of pathogenic microbes. As little is known about the interaction of biofloc systems and the addition of probiotics, this study focused on this integration to manipulate the microbial community and its interactions within biofloc systems. The present study evaluated two probiotics (B. velezensis AP193 and BiOWiSH FeedBuilder Syn 3) for use in Nile tilapia (Oreochromis niloticus) culture in a biofloc system. Nine independent 3785 L circular tanks were stocked with 120 juveniles (71.4 ± 4.4 g). Tilapia were fed for 16 weeks and randomly assigned three diets: a commercial control diet or a commercial diet top-coated with either AP193 or BiOWiSH FeedBuilder Syn3. At 14 weeks, the fish were challenged with a low dose of Streptococcus iniae (ARS-98-60, 7.2 × 107 CFU mL-1 , via intraperitoneal injection) in a common garden experimental design. At 16 weeks, the fish were challenged with a high dose of S. iniae (6.6 × 108 CFU mL-1 ) in the same manner. At the end of each challenge trial, cumulative per cent mortality, lysozyme activity and expression of 4 genes (il-1ß, il6, il8 and tnfα) from the spleen were measured. In both challenges, the mortalities of the probiotic-fed groups were significantly lower (p < .05) than in the control diet. Although there were some strong trends, probiotic applications did not result in significant immune gene expression changes related to diet during the pre-trial period and following exposure to S. iniae. Nonetheless, overall il6 expression was lower in fish challenged with a high dose of ARS-98-60, while tnfα expression was lower in fish subjected to a lower pathogen dose. Study findings demonstrate the applicability of probiotics as a dietary supplement for tilapia reared in biofloc systems.

Investigation of dietary exogenous protease and humic substance on growth, disease resistance to Flavobacterium covae and immune responses in juvenile channel catfish (Ictalurus punctatus).

Many bacterial pathogens impact the US catfish industry, and disease control can be challenging for producers. Columnaris disease in channel catfish, Ictalurus punctatus, is primarily caused by Flavobacterium covae (formerly F. columnare). Immunostimulants may enhance nonspecific immune responses and offer an alternative to antibiotic treatments in catfish. Furthermore, dietary protein sources and inclusions are also essential to fish health and nutrition and may enhance overall fish performance in pond culture. The current project evaluated two immunostimulants: a protease complex (PC) and a humic substance (HS) derived from a reed-sedge peat product. A 60-day trial examined the effects of five diets on growth performance, immune response and resistance to experimental F. covae infection in channel catfish. Diets included a high-quality fishmeal diet (32%; CF32), a high-protein soy-based diet (32%; C32), a low-protein soy-based diet (28%; C28; predominately used in industry), a low-protein soy diet supplemented with C28 + PC at 175 g metric ton-1 and C28 + HS in a low-protein diet at 23 g metric ton-1 . Following feeding for 60 d, juvenile channel catfish were sampled for growth performance and baseline health indicators (n = 3; body mucus, blood for sera, kidney and spleen). A subset of fish was then subjected to an immersion-based in vivo challenge trial with F. covae (ALG-00-530; 106 CFU mL-1 exposure). At 60d post-initiation, there were no dietary differences in the relative growth rate (p = .063) or thermal growth coefficient (p = .055), but the 32% diets generally appeared to perform best. Post-challenge, the C32 group's mortality was higher than the C28 + PC (p = .006) and C28 + HS diets (p = .005). Although not significant, the C28 and CF32 groups also demonstrated higher mortality compared to both PC and HS diets. Sera lysozyme concentration was found to increase following pathogen challenge (p < .001) and in comparison with mock-challenged catfish (p < .001). Elevated expression levels of proinflammatory cytokines (il-1ß, il-8, tnf-α and tgf-ß) were observed at trial midpoint and post-infection when compared to 60d. The C28 treatment was found to have lower tnf-α expression than the C28 + PC (p = .042) and C28 + HS (p = .042) groups following exposure to F. covae. These challenge data suggest that the immunostimulants (PC and HS) in plant-based protein may be beneficial in protecting against F. covae when offered in low-protein channel catfish diets.

Anxiety, stress, and depression in Australian pregnant women during the COVID-19 pandemic: A cross sectional study.

BACKGROUND: The COVID-19 pandemic necessitated rapid responses by health services to suppress transmission of the virus. AIM: This study aimed to investigate predictors of anxiety, stress and depression in Australian pregnant women during the COVID-19 pandemic including continuity of carer and the role of social support. METHODS: Women aged 18 years and over in their third trimester of pregnancy were invited to complete an online survey between July 2020 and January 2021. The survey included validated tools for anxiety, stress, and depression. Regression modelling was used to identify associations between a range of factors including continuity of carer, and mental health measures. FINDINGS: 1668 women completed the survey. One quarter screened positive for depression, 19% for moderate or higher range anxiety, and 15.5% for stress. The most significant contribution to higher anxiety, stress, and depression scores was a pre-existing mental health condition, followed by financial strain and a current complex pregnancy. Protective factors included age, social support, and parity. DISCUSSION: Maternity care strategies to reduce COVID-19 transmission restricted women's access to their customary pregnancy supports and increased their psychological morbidity. CONCLUSION: Factors associated with anxiety, stress and depression scores during the COVID-19 pandemic were identified. Maternity care during the pandemic compromised pregnant women's support systems.

Comparing an optimised physiotherapy treatment package with usual physiotherapy care for people with tennis elbow - protocol for the OPTimisE pilot and feasibility randomised controlled trial.

BACKGROUND: Physiotherapy is recommended for people with tennis elbow, but whilst a wide array of treatments is available, the optimal approach remains uncertain. We have therefore recently developed an optimised physiotherapy treatment package for tennis elbow based on a synthesis of the evidence, patient input and clinical consensus. It consists of detailed advice and education, a structured progressive exercise programme and provision of a counter-force elbow brace. Here, we report the protocol for our multicentre pilot and feasibility randomised controlled trial (RCT) designed to (a) examine the feasibility of our optimised physiotherapy treatment package and (b) to pilot trial processes for a future fully powered RCT to test clinical and cost-effectiveness compared with usual physiotherapy treatment. METHODS: A multicentre pilot and feasibility RCT will be conducted across three sites in England, recruiting up to 50 patients (or for a maximum of 12 months). Participants with tennis elbow, identified from physiotherapy clinic waiting lists and general practice surgeries, will be randomly allocated to receive the optimised physiotherapy treatment package or usual physiotherapy care. Analysis will focus on feasibility measures including consent rate, intervention fidelity, follow-up rate and outcome completion rate. A nested qualitative study will explore the acceptability of the study processes and patient and physiotherapist experiences of the new optimised intervention. DISCUSSION: This study will determine the feasibility of a new optimised physiotherapy treatment package for people with tennis elbow and pilot the processes for a future fully powered RCT. In the longer term, this treatment package may provide superior clinical outcomes for patients, in terms of pain and quality of life, and be more cost-effective for the health service. TRIAL REGISTRATION: Registered with the ISRCTN database 19/7/2021, https://www.isrctn.com/ISRCTN64444585.

Transcriptomic profiles of Florida pompano (Trachinotus carolinus) gill following infection by the ectoparasite Amyloodiniumocellatum.

The dinoflagellate Amyloodinium ocellatum is an important pathogenic parasite infecting cultured marine and brackish water fishes worldwide. This includes cultured Florida pompano (Trachinotus carolinus), which is one of the most desirable marine food fish with high economic value in the USA. A. ocellatum infects fish gills and causes tissue damage, increased respiratory rate, reduced appetite, and mortality, especially in closed aquaculture systems. This study mimicked the natural infection of A. ocellatum in cultured pompano and conducted a transcriptomic comparison of gene expression in the gills of control and A. ocellatum infected fish to explore the molecular mechanisms of infection. RNA-seq data revealed 604 differentially expressed genes in the infected fish gills. The immunoglobulin genes (including IgM/T) augmentation and IL1 inflammation suppression were detected after infection. Genes involved in reactive oxygen species mediating parasite killing were also highly induced. However, excessive oxidants have been linked to oxidative tissue damage and apoptosis. Correspondingly, widespread down-regulation of collagen genes and growth factor deprivation indicated impaired tissue repair, and meanwhile the key executor of apoptosis, caspase-3 was highly expressed (25.02-fold) in infected fish. The infection also influenced the respiratory gas sensing and transport genes and established hypoxic conditions in the gill tissue. Additionally, food intake and lipid metabolism were also affected. Our work provides the transcriptome sequencing of Florida pompano and provides key insights into the acute pathogenesis of A. ocellatum. This information can be utilized for designing optimal disease surveillance strategies, future selection for host resistance, and development of novel therapeutic measures.

Sources of variation in elemental profiles of whiteleg shrimp (Litopenaeus vannamei) and their potential effects on the accuracy of discriminant analysis.

BACKGROUND: Elemental profiling is a tool that has been proposed to improve the traceability of seafood products. Small sources of variation can affect the outcome of elemental profiling and therefore pose to lower the overall accuracy of analyses. Here, we investigate two potential sources of variation through three experiments: laboratory variation (intra-, interlaboratory variation, and tissue matrix) and tissue variation. METHODS AND RESULTS: Samples of whitleleg shrimp (Litopenaeus vannamei) were obtained from 20 farms in Ecuador and two farms in Alabama to be analyzed. In the first experiment of the study, samples from Ecuador were analyzed at three different laboratories and compared. Two out of the five elements reported were statistically different across the three laboratories (Cu and Se). In the second experiment, the effect of tissue matrix (ground vs whole tissue during acid digestion) was investigated. Altogether, five out of 29 elements analyzed were statistically different. In the third experiment, samples from two farms in Alabama were analyzed to understand the variation in element concentrations in different tissues (head on shell on (HOSO), headless shell on (HLSO), headless peeled (PLD) and headless peeled and deveined (PLDV)). Elemental concentrations varied across tissues, and patterns in elemental concentrations were site specific. The samples from the two farms were analyzed with a Random Forest classification model to site x tissue groupings with 94% accuracy. CONCLUSION: The result of this study highlights the following: 1. Consistency in laboratory analysis important in studies that involve element concentrations, as minor differences in methodology can propagate as significant differences in results. 2. In shrimp, elements are compartmentalized in different tissues and elemental profiling should consistently use the same type of tissue.

Tobacco Use and Tobacco Treatment Referral Response of Patients With Cancer: Implementation Outcomes at a National Cancer Institute-Designated Cancer Center.

PURPOSE: Smoking after a cancer diagnosis is linked to cancer-specific and all-cause mortality, among other adverse outcomes. Yet, 10%-20% of US cancer survivors are current smokers. Implementation of evidence-based tobacco treatment in cancer care facilities is widely recommended, yet rarely accomplished. This study focuses on the early outcomes of a population-based tobacco treatment program integrated within an National Cancer Institute-designated cancer center. METHODS AND MATERIALS: The sample consists of 26,365 patients seen at the cancer center during the first 18 months of program implementation. The study is a retrospective chart review of patients' tobacco use and, among current users, patients' treatment referral response. RESULTS: More than 99% of patients were screened for tobacco use. Current (past month) use was observed in 21.05% of patients; cigarettes were the most popular product. Only 17.22% of current users accepted a referral for tobacco treatment; among current users who declined, the majority were not ready to quit (65.84%) or wanted to quit on their own (27.01%). Multiple demographic variables were associated with tobacco use and treatment referral response outcomes. CONCLUSION: Despite cancer diagnosis presenting a teachable moment for tobacco cessation, patients with cancer may not be ready to quit or engage with treatment. Clinically proven strategies to increase motivation, prompt quit attempts, and encourage treatment use should be key components of tobacco treatment delivery to patients with cancer.

Point Absorber Limits to Future Gravitational-Wave Detectors.

High-quality optical resonant cavities require low optical loss, typically on the scale of parts per million. However, unintended micron-scale contaminants on the resonator mirrors that absorb the light circulating in the cavity can deform the surface thermoelastically and thus increase losses by scattering light out of the resonant mode. The point absorber effect is a limiting factor in some high-power cavity experiments, for example, the Advanced LIGO gravitational-wave detector. In this Letter, we present a general approach to the point absorber effect from first principles and simulate its contribution to the increased scattering. The achievable circulating power in current and future gravitational-wave detectors is calculated statistically given different point absorber configurations. Our formulation is further confirmed experimentally in comparison with the scattered power in the arm cavity of Advanced LIGO measured by in situ photodiodes. The understanding presented here provides an important tool in the global effort to design future gravitational-wave detectors that support high optical power and thus reduce quantum noise.

A preliminary survey of antibiotic residues in frozen shrimp from retail stores in the United States.

Shrimp are an important and valuable commodity for aquaculture that are widely traded internationally. Widespread antibiotic use has been documented in shrimp farming and is a common source of criticism of aquaculture products. Additionally, previous reports have found some evidence of antibiotic residues in shrimp samples obtained from retail stores in the United States, which is a concern for consumers. To further understand the prevalence of antibiotics in retail shrimp in the United States, shrimp samples obtained from grocery stores across 16 states were analyzed for 74 antibiotic compounds/metabolites at a commercial laboratory. 68 samples were analyzed for a multiclass antibiotic panel which included 66 antibiotics while a subset of 15 samples were analyzed for ß-lactam antibiotics, Nitrofurans, and Oxytetracycline. Samples were obtained that were labeled as being from major production countries, including India, Indonesia, Thailand, and Vietnam. No detectable antibiotic residues were found in this survey in any samples. This is contrary to previous findings in frozen shrimp analyzed for antibiotics, which typically report low levels of the prevalence of antibiotics.

Trace element concentrations in white leg shrimp Litopenaeus vannamei from retail stores in the EU, UK, and USA and the ability to discern country of origin with classification models.

Shrimp are a globally traded aquaculture commodity that accounts for a large proportion of the monetary value of aquaculture. There are concerns among consumers about seafood labeling fraud and environmental sustainability. Therefore, the geographic origin of shrimp from retail stores was investigated with trace element profiling. 94 shrimp samples were collected from grocery stores across the USA, UK, and EU in 70 different grocery stores. The results of 24 elements are reported. Shrimp samples were from Thailand, India, Vietnam, Indonesia, and Ecuador were shown to have 15 elements that were statistically different across labeled country of origin, with Ecuador having unique post hoc group membership in 5 of the elements. Based on a classification procedure, shrimp were classified to labeled country of origin with an overall accuracy of 71.2%. Overall, the results suggest that elemental profiling could be a traceability tool for classifying samples of shrimp from retail stores.

Effect of salinity on growth, survival, and serum osmolality of red snapper, Lutjanus campechanus.

Three trials were conducted to evaluate the performances of red snapper, Lutjanus campechanus, in low salinities. The median lethal concentration (96 h LC50) of salinity was determined by trimmed Spearman-Karber method using survival data of fish (18.9 ± 0.2 g) collected after 96 h from acclimation to 2, 4, 8, and 32 ppt salinities in 800 L tanks (n = 3), while the serum osmolality of fish (74.1 ± 3.9 g) was determined after 48 h from acclimation to 6, 8, 16, 24, and 32 ppt salinities in 150 L tanks (n = 3). The growth trial was conducted for 6 weeks in 800 L tanks to determine the growth and survival of fish (18.8 ± 0.2 g) at 8 ppt salinity compared to the control (32 ppt salinity). At the conclusion, the isosmotic point of fish was estimated as 357.2 mmol/kg (correspond to 11.0 ppt salinity), while the 96 h LC50 was estimated as 5.65 ppt salinity. No significant differences were noted for survival and FCR of fish reared in 8 and 32 ppt salinities. However, growth was significantly lower in fish reared in 8 ppt salinity compared to the fish reared in 32 ppt salinity. The reduced growth could be, at least partially, due to the increased osmoregulatory energy expenditure at lower salinities.

Performance characteristics and costs of serological tests for brucellosis in a pastoralist community of northern Tanzania.

The control of brucellosis across sub-Saharan Africa is hampered by the lack of standardized testing and the use of tests with poor performance. This study evaluated the performance and costs of serological assays for human brucellosis in a pastoralist community in northern Tanzania. Serum collected from 218 febrile hospital patients was used to evaluate the performance of seven index tests, selected based on international recommendation or current use. We evaluated the Rose Bengal test (RBT) using two protocols, four commercial agglutination tests and a competitive enzyme-linked immunosorbent assay (cELISA). The sensitivity, specificity, positive predictive value, negative predictive value, Youden's index, diagnostic accuracy, and per-sample cost of each index test were estimated. The diagnostic accuracy estimates ranged from 95.9 to 97.7% for the RBT, 55.0 to 72.0% for the commercial plate tests, and 89.4% for the cELISA. The per-sample cost range was $0.69-$0.79 for the RBT, $1.03-$1.14 for the commercial plate tests, and $2.51 for the cELISA. The widely used commercial plate tests performed poorly and cost more than the RBT. These findings provide evidence for the public health value of discontinuing the use of commercial agglutination tests for human brucellosis in Tanzania.

Growth, serum biochemical parameters, salinity tolerance and antioxidant enzyme activity of rainbow trout (Oncorhynchus mykiss) in response to dietary taurine levels.

This study evaluated the effect of dietary taurine levels on growth, serum biochemical parameters, salinity adaptability, and antioxidant activity of rainbow trout (Oncorhynchus mykiss). Four diets were formulated with taurine supplements at 0, 0.5, 1, and 2% w/v (abbreviated as T0, T0.5, T1, and T2, respectively). Rainbow trouts (initial weight of 80.09 ± 4.72 g) were stocked in tanks (180 L capacity), and were fed these diets for six weeks and subsequently underwent salinity acclimation. Physiological indicators were determined before salinity acclimation at 1, 4, 7, and 14 days afterwards. Results showed that there were no significant differences in growth performance (final mean weight ranged from 182.35 g to 198.48 g; percent weight gain was between 127.68% and 147.92%) of rainbow trout in freshwater stage, but dietary taurine supplement significantly increased serum-free taurine content. After entering seawater, the Na+-K+-ATPase activity of T2 group returned to its freshwater levels, and the serum cortisol content was significantly higher than T0 and T0.5 groups. At the end of this experiment, the liver superoxide dismutase activity in the T0 and T0.5 groups was significantly lower than in the T1 and T2 groups, and the liver catalase in the T0 group was the lowest whereas that in the T2 group was the highest. Muscle malondialdehyde content was the highest in the T0 group, and the lowest in the T2 group. Based on the results of this study, supplement of dietary taurine (0.5-2%) enhanced the salinity tolerance in rainbow trout, which increased with the higher taurine concentration.

Effects of Ground Transport in Kemp's Ridley (Lepidochelys kempii) and Loggerhead (Caretta caretta) Turtles.

Many juvenile Kemp's ridley (Lepidochelys kempii) and loggerhead (Caretta caretta) turtles strand during fall on the beaches of Cape Cod (MA, USA), with total stranding numbers sometimes exceeding 300 turtles per year. Once rehabilitated, turtles must be released at beaches with appropriate water temperatures, often requiring transportation to southeastern coastal states of the USA. These transportation events (transports) may approach or exceed 24 h in duration. Kemp's ridley turtles are known to exhibit an adrenal stress response during such transports, but the effect of transport duration has been unclear, and no other sea turtle species has been investigated. To assess whether transport duration and/or species affects physiological reactions to transport, we studied pre- and post-transport physiological measures in Kemp's ridley and loggerhead turtles transported by ground for <6, ∼12, ∼18, or ∼24 h, comparing with matched "control events" in which turtles were studied without transport. Blood samples were analyzed for four stress-associated measures (corticosterone, glucose, total white blood cell [WBC] count, and heterophil/lymphocyte ratio [H/L]) and nine measures of clinical status (pH, pO2, pCO2, HCO3, sodium, potassium, ionized calcium, lactate, and hematocrit). In both species, stress-associated measures elevated significantly during transport, while handling without transport had no significant effects. Loggerheads exhibited a greater stress response than Kemp's ridleys across all transport durations. These results indicate that sea turtles do react physiologically to ground transport; therefore, minimizing transport time and streamlining transport logistics (where feasible) may help ensure release of rehabilitated turtles to sea in the best possible condition. Nonetheless, both species remained in good clinical condition even after 24 h transport, indicating that current transport protocols are generally safe for sea turtles from a clinical perspective.

A European multicentre evaluation of detection and typing methods for human enteroviruses and parechoviruses using RNA transcripts.

Polymerase chain reaction (PCR) detection has become the gold standard for diagnosis and typing of enterovirus (EV) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using the appropriate sample types and high assay sensitivity as viral loads in cerebrospinal fluid samples from meningitis and sepsis clinical presentation can be extremely low. This study evaluated the sensitivity and specificity of currently used commercial and in-house diagnostic and typing assays. Accurately quantified RNA transcript controls were distributed to 27 diagnostic and 12 reference laboratories in 17 European countries for blinded testing. Transcripts represented the four human EV species (EV-A71, echovirus 30, coxsackie A virus 21, and EV-D68), HPeV3, and specificity controls. Reported results from 48 in-house and 15 commercial assays showed 98% detection frequencies of high copy (1000 RNA copies/5 µL) transcripts. In-house assays showed significantly greater detection frequencies of the low copy (10 copies/5 µL) EV and HPeV transcripts (81% and 86%, respectively) compared with commercial assays (56%, 50%; P = 7 × 10-5 ). EV-specific PCRs showed low cross-reactivity with human rhinovirus C (3 of 42 tests) and infrequent positivity in the negative control (2 of 63 tests). Most or all high copy EV and HPeV controls were successfully typed (88%, 100%) by reference laboratories, but showed reduced effectiveness for low copy controls (41%, 67%). Stabilized RNA transcripts provide an effective, logistically simple and inexpensive reagent for evaluation of diagnostic assay performance. The study provides reassurance of the performance of the many in-house assay formats used across Europe. However, it identified often substantially reduced sensitivities of commercial assays often used as point-of-care tests.