Radiographic characterisation of spinal curvature development in farmed New Zealand Chinook salmon Oncorhynchus tshawytscha throughout seawater production.

Spinal anomalies are a recognised source of downgrading in finfish aquaculture, but identifying their cause(s) is difficult and often requires extensive knowledge of the underlying pathology. Late-onset spinal curvatures (lordosis, kyphosis, scoliosis) can affect up to 40% of farmed New Zealand Chinook (king) salmon (Oncorhynchus tshawytscha) at harvest, but little is known about their pathogenesis. Curvature development was radiographically documented in two related cohorts of commercially-farmed Chinook salmon throughout seawater production to determine (1) the timing of radiographic onset and relationships between (2) the curvature types, (3) the spinal regions in which they develop and (4) their associations with co-existing vertebral body anomalies (vertebral compression, fusion and vertical shift). Onset of curvature varied between individuals, but initially occurred eight months post-seawater transfer. There were strong associations between the three curvature types and the four recognised spinal regions: lordosis was predominantly observed in regions (R)1 and R3, kyphosis in R2 and R4, manifesting as a distinct pattern of alternating lordosis and kyphosis from head to tail. This was subsequently accompanied by scoliosis, which primarily manifested in spinal regions R2 and R3, where most of the anaerobic musculature is concentrated. Co-existing vertebral body anomalies, of which vertebral compression and vertical shift were most common, appeared to arise either independent of curvature development or as secondary effects. Our results suggest that spinal curvature in farmed New Zealand Chinook salmon constitutes a late-onset, rapidly-developing lordosis-kyphosis-scoliosis (LKS) curvature complex with a possible neuromuscular origin.

Potential of genomic selection for improvement of resistance to ostreid herpesvirus in Pacific oyster (Crassostrea gigas).

In genomic selection (GS), genome-wide SNP markers are used to generate genomic estimated breeding values for selection candidates. The application of GS in shellfish looks promising and has the potential to help in dealing with one of the main issues currently affecting Pacific oyster production worldwide, which is the 'summer mortality syndrome'. This causes periodic mass mortality in farms worldwide and has mainly been attributed to a specific variant of the ostreid herpesvirus (OsHV-1). In the current study, we evaluated the potential of genomic selection for host resistance to OsHV-1 in Pacific oysters, and compared it with pedigree-based approaches. An OsHV-1 disease challenge was performed using an immersion-based virus exposure treatment for oysters for 7 days. A total of 768 samples were genotyped using the medium-density SNP array for oysters. A GWAS was performed for the survival trait using a GBLUP approach in blupf90 software. Heritability ranged from 0.25 ± 0.05 to 0.37 ± 0.05 (mean ± SE) based on pedigree and genomic information respectively. Genomic prediction was more accurate than pedigree prediction, and SNP density reduction had little impact on prediction accuracy until marker densities dropped below approximately 500 SNPs. This demonstrates the potential for GS in Pacific oyster breeding programmes, and importantly, demonstrates that a low number of SNPs might suffice to obtain accurate genomic estimated breeding values, thus potentially making the implementation of GS more cost effective.

Assessment of genetic diversity and population structure in cultured Australian Pacific oysters.

The recent development of Pacific oyster (Crassostrea gigas) SNP genotyping arrays has allowed detailed characterisation of genetic diversity and population structure within and between oyster populations. It also raises the potential of harnessing genomic selection for genetic improvement in oyster breeding programmes. The aim of this study was to characterise a breeding population of Australian oysters through genotyping and analysis of 18 027 SNPs, followed by comparison with genotypes of oyster sampled from Europe and Asia. This revealed that the Australian populations had similar population diversity (HE ) to oysters from New Zealand, the British Isles, France and Japan. Population divergence was assessed using PCA of genetic distance and revealed that Australian oysters were distinct from all other populations tested. Australian Pacific oysters originate from planned introductions sourced from three Japanese populations. Approximately 95% of these introductions were from geographically, and potentially genetically, distinct populations from the Nagasaki oysters assessed in this study. Finally, in preparation for the application of genomic selection in oyster breeding programmes, the strength of LD was evaluated and subsets of loci were tested for their ability to accurately infer relationships. Weak LD was observed on average; however, SNP subsets were shown to accurately reconstitute a genomic relationship matrix constructed using all loci. This suggests that low-density SNP panels may have utility in the Australian population tested, and the findings represent an important first step towards the design and implementation of genomic approaches for applied breeding in Pacific oysters.

Epithelial Cell Lineage and Signaling in Murine Salivary Glands.

Maintaining salivary gland function is critical for oral health. Loss of saliva is a common side effect of therapeutic irradiation for head and neck cancer or autoimmune diseases such as Sjögren's syndrome. There is no curative treatment, and current strategies proposed for functional regeneration include gene therapy to reengineer surviving salivary gland tissue, cell-based transplant therapy, use of bioengineered glands, and development of drugs/biologics to stimulate in vivo regeneration or increase secretion. Understanding the genetic and cellular mechanisms required for development and homeostasis of adult glands is essential to the success of these proposed treatments. Recent advances in genetic lineage tracing provide insight into epithelial lineage relationships during murine salivary gland development. During early fetal gland development, epithelial cells expressing keratin 14 (K14) Sox2, Sox9, Sox10, and Trp63 give rise to all adult epithelium, but as development proceeds, lineage restriction occurs, resulting in separate lineages of myoepithelial, ductal, and acinar cells in postnatal glands. Several niche signals have been identified that regulate epithelial development and lineage restriction. Fibroblast growth factor signaling is essential for gland development, and other important factors that influence epithelial patterning and maturation include the Wnt, Hedgehog, retinoic acid, and Hippo signaling pathways. In addition, other cell types in the local microenvironment, such as endothelial and neuronal cells, can influence epithelial development. Emerging evidence also suggests that specific epithelial cells will respond to different types of salivary gland damage, depending on the cause and severity of damage and the resulting damaged microenvironment. Understanding how regeneration occurs and which cell types are affected, as well as which signaling factors drive cell lineage decisions, provides specific targets to manipulate cell fate and improve regeneration. Taken together, these recent advances in understanding cell lineages and the signaling factors that drive cell fate changes provide a guide to develop novel regenerative treatments.

Prevalence of spinal abnormalities in Chinook salmon smolt and influence of early rearing temperature and growth rates.

Spinal abnormalities can be detected at harvest in around 40% of farmed Chinook salmon in New Zealand. However, whether these abnormalities are present in smolt is unknown. Radiographs of 3,736 smolt were taken immediately prior to transfer to sea water and evaluated for fusions, compressions, vertical shifts, and lordosis, kyphosis and/or scoliosis (LKS). The survey included smolt from two different chilling strategies that had been graded into slow- or fast-growing fish. Overall, 4.34% of Chinook salmon smolt had at least one spinal abnormality, similar to the rates of reported in Atlantic salmon smolt. The rate of abnormality was significantly higher in faster-growing fish. Fusions were most common with 2.68% of smolt affected. Smolt subjected to longer chilling times had lower rates of fusions. Compressions and vertical shifts were both observed in 1.31% of smolt. Although LKS is the most common abnormality of harvested fish, LKS was detected in just five smolt. The results suggest that some fusions in harvest fish have developed at the time of seawater transfer while LKS develops late in the production cycle. Overall, spinal abnormalities are uncommon in Chinook salmon smolt and may be influenced by chilling times and growth rates.

Use of a saliva-based diagnostic test to identify tapeworm infection in horses in the UK.

BACKGROUND: Anthelmintic resistance combined with limited chemotherapeutic options has prompted a change in approaches to control of equine helminth infections. Targeted selective treatment strategies use diagnostics to reduce anthelmintic use by treating individuals with worm burdens or egg shedding levels above a set threshold. While faecal egg count analysis has limitations for informing tapeworm treatment, a commercially available saliva-based diagnostic test accurately diagnoses horses with tapeworm infection. OBJECTIVES: Evaluation of a saliva-based diagnostic test to identify horses naturally infected with tapeworm and assess the impact of using the test to inform anthelmintic administration. STUDY DESIGN: Retrospective longitudinal study. METHODS: Saliva was collected from horses (n = 237) at a UK welfare charity from autumn 2015 to autumn 2016. Horses diagnosed as positive for tapeworm infection using the EquiSal® Tapeworm test were anthelmintic treated according to weight. The number of horses that received anthelmintic treatment based on the test result was compared with an all-group treatment approach and the reduction in anthelmintic usage calculated. Incoming horses were also tested (n = 143) and the information was used to inform quarantine treatments. RESULTS: In autumn 2015, 85% of 237 horses tested received no anthelmintic and the majority (71%) of these remained below the treatment threshold throughout the study. Of the 69 horses that received treatment, seven required treatment following three subsequent tests, while >50% of horses administered with anthelmintic fell below the treatment threshold at the following test. No increase in tapeworm prevalence within the 237 horses was observed during the study despite a substantial reduction in the application of antitapeworm treatments. A total of 41% of incoming horses required anticestode treatment. MAIN LIMITATIONS: Other management practices were not included in the analysis. CONCLUSIONS: Compared with an all-group treatment strategy, the diagnostic-led approach used here considerably reduced application of anticestode anthelmintics. This could reduce selection pressure for anthelmintic resistance.

Glycolysis inhibition improves photodynamic therapy response rates for equine sarcoids.

Photodynamic therapy (PDT) holds great promise in treating veterinary and human dermatological neoplasms, including equine sarcoids, but is currently hindered by the amount of photosensitiser and light that can be delivered to lesions thicker than around 2 mm, and by the intrinsic antioxidant defences of tumour cells. We have developed a new PDT technique that combines an efficient transdermal penetration enhancer solution, for topical delivery of 5-aminolevulinic acid (ALA) photosensitiser, with acute topical post-PDT application of the glycolysis inhibitor lonidamine. We show that the new PDT combination treatment selectively kills sarcoid cells in vitro, with repeated rounds of treatment increasing sarcoid sensitisation to PDT. In vivo, ALA PDT followed by 600 µM lonidamine substantially improves treatment outcomes for occult, verrucous, nodular and fibroblastic sarcoids after 1 month (93% treatment response in 27 sarcoids), compared with PDT using only ALA (14% treatment response in 7 sarcoids).

The relationship between interfacial bonding and radiation damage in adsorbed DNA.

We have performed a comparison of the radiation damage occurring in DNA adsorbed on gold in two different configurations, when the DNA is thiolated and bound covalently to the substrate and when it is unthiolated and interacts with the substrate through the bases. Both molecules were found to organize so as to protrude from the surface at ~45 degrees. Changes in the time-dependent C 1s and O 1s X-ray photoelectron (XP) spectra resulting from irradiation were interpreted to arise from cleavage of the phosphodiester bond and possibly COH desorption. By fitting the time-dependent XP spectra to a simple kinetic model, time constants were extracted, which were converted to cross sections and quantum yields for the damage reaction. The radiation induced damage is significantly higher for the thiolated DNA. N 1s X-ray absorption spectrum revealed the N-C=N LUMO is more populated in the unthiolated molecule, which is due to a higher degree of charge transfer from the substrate to this LUMO in the unthiolated case. Since the N-C=N LUMO of the thiolated molecule is comparatively less populated, it is more effective in capturing low energy electrons resulting in a higher degree of damage.

Optimised parent selection and minimum inbreeding mating in small aquaculture breeding schemes: a simulation study.

The effectiveness of low cost breeding scheme designs for small aquaculture breeding programmes were assessed for their ability to achieve genetic gain while managing inbreeding using stochastic simulation. Individuals with trait data were simulated over 15 generations with selection on a single trait. Combinations of selection methods, mating strategies and genetic evaluation options were evaluated with and without the presence of common environmental effects. An Optimal Parent Selection (OPS) method using semi-definite programming was compared with a truncation selection (TS) method. OPS constrains the rate of inbreeding while maximising genetic gain. For either selection method, mating pairs were assigned from the selected parents by either random mating (RM) or Minimum Inbreeding Mating (MIM), which used integer programming to determine mating pairs. Offspring were simulated for each mating pair with equal numbers of offspring per pair and these offspring were the candidates for selection of parents of the next generation. Inbreeding and genetic gain for each generation were averaged over 25 replicates. Combined OPS and MIM led to a similar level of genetic gain to TS and RM, but inbreeding levels were around 75% lower than TS and RM after 15 generations. Results demonstrate that it would be possible to manage inbreeding over 15 generations within small breeding programmes comprised of 30 to 40 males and 30 to 40 females with the use of OPS and MIM. Selection on breeding values computed using Best Linear Unbiased Prediction (BLUP) with all individuals genotyped to obtain pedigree information resulted in an 11% increase in genetic merit and a 90% increase in the average inbreeding coefficient of progeny after 15 generations compared with selection on raw phenotype. Genetic evaluation strategies using BLUP wherein elite individuals by raw phenotype are genotyped to obtain parentage along with a range of different samples of remaining individuals did not increase genetic progress in comparison to selection on raw phenotype. When common environmental effects on full-sib families were simulated, performance of small breeding scheme designs was little affected. This was because the majority of selection must anyway be applied within family due to inbreeding constraints.

Early development of New Zealand hapuku Polyprion oxygeneios eggs and larvae.

This study describes for the first time the normal development of New Zealand hapuku Polyprion oxygeneios embryos and larvae reared from fertilization to 11 days post-hatch (dph) at a constant temperature. Fertilized eggs were obtained from natural spawnings from communally reared captive wild broodstock. Eggs averaged 2 mm in diameter and had single or multiple oil globules. Embryos developed following the main fish embryological stages and required an average of 1859·50 degree hours post-fertilization (dhpf) to hatch. The newly hatched larvae (4·86 mm mean total length, L(T) ) were undifferentiated, with unpigmented eyes, a single and simple alimentary tube and a finfold that covered the entire body. Larvae relied on the energy from the yolk-sac reserves until 11 dph (7·33 mm mean L(T) ), when yolk-sac reabsorption was almost completed. Some of the major developmental stages from hatching to yolk-sac reabsorption were eye pigmentation (5 dph), upper jaw formation (7 dph), lower jaw formation (8 dph) and mouth opening (8-9 dph). By 9 dph, the digestive system consisted of pancreas, liver, primordial stomach, anterior and posterior gut; therefore, P. oxygeneios larvae would be capable of feeding on live prey. The developmental, morphological and histological data described constitutes essential baseline information on P. oxygeneios biology and normal development.

Immediate and associated complications of hysterectomy for benign disease.

Abdominal and vaginal hysterectomy are common operations for benign gynaecological conditions. Though safe operations, they are not without complications. Good medical practice requires that patients be fully counselled regarding risks involved in undergoing medical interventions. This can only be done by regular review of local practice and comparison with national and international standards. We have reviewed 502 case notes of patients who have undergone a hysterectomy for benign conditions over an 18-month period.

Identification of an occludin cell adhesion recognition sequence.

The molecular mechanisms by which the tight junction integral membrane protein, occludin promotes cell adhesion and establishes an endothelial monolayer permeability barrier have not been elucidated. In particular, the amino acid sequences of the occludin cell adhesion recognition (CAR) sites have not been determined. Here we demonstrate that a cyclic peptide containing the sequence LYHY, which is found in the second extracellular domain of occludins in all mammalian species, inhibits the establishment of endothelial cell barriers in vitro and in vivo. This cyclic peptide also prevents the aggregation of fibroblasts stably transfected with cDNA encoding occludin. The data suggest that the LYHY motif is an occludin CAR sequence.

Calreticulin affects beta-catenin-associated pathways.

Calreticulin, a Ca(2+) storage protein and chaperone in the endoplasmic reticulum, also modulates cell adhesiveness. Overexpression of calreticulin correlates with (i) increased cell adhesiveness, (ii) increased expression of N-cadherin and vinculin, and (iii) decreased protein phosphorylation on tyrosine. Among proteins that are dephosphorylated in cells that overexpress calreticulin is beta-catenin, a structural component of cadherin-dependent adhesion complexes, a member of the armadillo family of proteins and a part of the Wnt signaling pathway. We postulate that the changes in cell adhesiveness may be due to calreticulin-mediated effects on a signaling pathway from the endoplasmic reticulum, which impinges on the Wnt signaling pathway via the cadherin/catenin protein system and involves changes in the activity of protein-tyrosine kinases and/or phosphatases.

Electroporation of salmon sperm for gene transfer: efficiency, reliability, and fate of transgene.

Uptake of exogenous DNA by electroporated salmon sperm for gene transfer is being investigated. Our studies show that electroporated salmon sperm cells were more efficient and more reliable than untreated sperm in picking up exogenous DNA and subsequently transferring the DNA into salmon embryos. Indirect evidence suggest that some of the exogenous DNA was internalized in the sperm nuclei. The taken up DNA retained its integrity as demonstrated by PCR. The foreign DNA was detected in 15-month-old fish, and had a mosaic pattern of distribution. Integration of the foreign DNA occurred infrequently, and the expression of the foreign genes was poor. The potential of sperm-mediated gene transfer as a routine protocol for mass gene transfer in salmon will be dependent on the improvement of integration and expression of the foreign gene.

Cadherins: crucial regulators of structure and function in reproductive tissues.

Cadherins are cell surface proteins that are directly involved in a wide variety of processes such as cell adhesion, cell sorting, cell survival, morphogenesis, formation of intercellular junctions, maintenance of tissue integrity and tumourigenesis. This review discusses the multiple functions of cadherins in reproductive tissues. Furthermore, the role of the intracellular signalling protein beta-catenin in regulating cadherin function is reviewed. Finally, the findings that cadherin concentrations in reproductive tissues are responsive to steroid hormones is discussed. The modulation of cadherin expression by hormones is in agreement with the hypothesis that these proteins are dynamically involved in the maintenance of structure and function in reproductive tissues.

Pyomyositis: an unusual infection due to staphylococcus aureus.

Pyomyositis is a primary pyogenic infection of skeletal muscle, leading to the formation of intramuscular abscesses. Although common in tropical climates, it is infrequent in temperate zones. We report a patient who developed the condition without travelling to tropical areas.