Free and open-source software for object detection, size, and colour determination for use in plant phenotyping.

BACKGROUND: Object detection, size determination, and colour detection of images are tools commonly used in plant science. Key examples of this include identification of ripening stages of fruit such as tomatoes and the determination of chlorophyll content as an indicator of plant health. While methods exist for determining these important phenotypes, they often require proprietary software or require coding knowledge to adapt existing code. RESULTS: We provide a set of free and open-source Python scripts that, without any adaptation, are able to perform background correction and colour correction on images using a ColourChecker chart. Further scripts identify objects, use an object of known size to calibrate for size, and extract the average colour of objects in RGB, Lab, and YUV colour spaces. We use two examples to demonstrate the use of these scripts. We show the consistency of these scripts by imaging in four different lighting conditions, and then we use two examples to show how the scripts can be used. In the first example, we estimate the lycopene content in tomatoes (Solanum lycopersicum) var. Tiny Tim using fruit images and an exponential model to predict lycopene content. We demonstrate that three different cameras (a DSLR camera and two separate mobile phones) are all able to model lycopene content. The models that predict lycopene or chlorophyll need to be adjusted depending on the camera used. In the second example, we estimate the chlorophyll content of basil (Ocimum basilicum) using leaf images and an exponential model to predict chlorophyll content. CONCLUSION: A fast, cheap, non-destructive, and inexpensive method is provided for the determination of the size and colour of plant materials using a rig consisting of a lightbox, camera, and colour checker card and using free and open-source scripts that run in Python 3.8. This method accurately predicted the lycopene content in tomato fruit and the chlorophyll content in basil leaves.

Space controlled environment agriculture offers pathways to improve the sustainability of controlled environmental agriculture on Earth.

Terrestrial controlled environment agriculture (CEA) will have an increasingly important role in food production. However, present CEA systems are energy- and resource-hungry and rarely profitable, requiring a step change in design and optimization. Here we argue that the unique nature of space controlled environment agriculture (SpaCEA), which needs to be both highly resource efficient and circular in design, presents an opportunity to develop intrinsically circular CEA systems. Life-cycle analysis tools should be used to optimize the provision and use of natural or electrical light, power, nutrients and infrastructure in CEA and/or SpaCEA systems, and to guide research and development into subsystems that bring strong environmental advantages. We suggest that SpaCEA public outreach can also be used to improve the perception of terrestrial CEA on Earth by using space as a gateway for exhibiting CEA food growing technologies. A substantial focus on SpaCEA development should be viewed as an efficient contribution to addressing major current CEA challenges.

Carbon Dioxide Laser Rejuvenation of the Facial Skin.

This article reviews the evaluation and techniques for facial skin rejuvenation using the fractionated carbon dioxide laser. It includes a detailed overview of laser skin rejuvenation and discusses the potential complications associated with this procedure. A review of clinical outcomes in the literature is also included.

Rational Design of a Polyurethane Foam.

Polyurethane (PU) foams are exceptionally versatile due to the nature of PU bond formation and the large variety of polymeric backbones and formulation components such as catalysts and surfactants. This versatility introduces a challenge, namely a near unlimited number of variables for formulating foams. In addition to this, PU foam development requires expert knowledge, not only in polyurethane chemistry but also in the art of evaluating the resulting foams. In this work, we demonstrate that a rational experimental design framework in conjunction with a design of experiments (DoE) approach reduces both the number of experiments required to understand the formulation space and reduces the need for tacit knowledge from a PU expert. We focus on an in-depth example where a catalyst and two surfactants of a known formulation are set as factors and foam physical properties are set as responses. An iterative DoE approach is used to generate a set of foams with substantially different cell morphology and hydrodynamic behaviour. We demonstrate that with 23 screening formulations and 16 final formulations, foam physical properties can be modelled from catalyst and surfactant loadings. This approach also allows for the exploration of relationships between the cell morphology of PU foam and its hydrodynamic behaviour.

FoamPi: An open-source raspberry Pi based apparatus for monitoring polyurethane foam reactions.

Adiabatic temperature rise is an important method for determining isocyanate conversion in polyurethane foam reactions as well as many other exothermic chemical reactions. Adiabatic temperature rise can be used in conjunction with change in height and mass measurements to gain understanding into the blowing and gelling reactions that occur during polyurethane foaming as well as give important information on cell morphology. FoamPi is an open-source Raspberry Pi device for monitoring polyurethane foaming reactions. The device effectively monitors temperature rise, change in foam height as well as changes in the mass during the reaction. Three Python scripts are also presented. The first logs raw data during the reaction. The second corrects temperature data such that it can be used in adiabatic temperature rise reactions for calculating isocyanate conversion; additionally this script reduces noise in all the data and removes erroneous readings. The final script extracts important information from the corrected data such as maximum temperature change and maximum height change as well as the time to reach these points. Commercial examples of such equipment exist however the price (>£10000) of these equipment make these systems inaccessible for many research laboratories. The FoamPi build presented is inexpensive (£350) and test examples are shown here to indicate the reproducibility of results as well as precision of the FoamPi.

Carbon Dioxide Laser Rejuvenation of the Facial Skin.

This article reviews the evaluation and techniques for facial skin rejuvenation using the fractionated carbon dioxide laser. It includes a detailed overview of laser skin rejuvenation and discusses the potential complications associated with this procedure. A review of clinical outcomes in the literature is also included.

Differences in the protection elicited by a recombinant Teladorsagia circumcincta vaccine in weaned lambs of two Canarian sheep breeds.

Gastrointestinal nematode (GIN) infections are a serious drawback on small ruminant production. Since anthelmintic resistance has extended, optimisation of alternative non-chemical control strategies has attracted interest. Recently, a prototype recombinant vaccine protected immunologically mature sheep from Texel-cross and Canaria Sheep breeds against Teladorsagia circumcincta. The level of protective immunity stimulated by the vaccine varied between individuals and with age. Previous studies suggest that Canaria Hair Breed (CHB) sheep is naturally resistant to GIN infection, with some evidence suggesting that this protection is present in young lambs. Here, we sought to enhance this resistance by immunising three-month-old CHB lambs with a T. circumcincta prototype recombinant vaccine. Following vaccination and a larval challenge period, levels of protection against T. circumcincta infection were compared in CHB lambs with Canaria Sheep (CS) lambs (a breed considered less resistant to GIN). Lambs from the resistant CHB breed appeared to respond more favourably to vaccination, shedding 63% fewer eggs over the sampling period than unvaccinated CHB lambs. No protection was evident in CS vaccinated lambs. At post-mortem, CHB vaccine recipients had a 68% reduction in mean total worm burden, and female worms were significantly shorter and contained fewer eggs in utero compared to unvaccinated CHB lambs. A higher anti-parasite IgG2 level was detected in immunised CHB lambs compared to unvaccinated control CHB animals, with data suggesting that IgA, globular leucocytes, CD45RA+, CD4+ and CD8+ T cells are implicated in this protective response. The development of effective immunity in vaccinated CHB lambs did not reduce lamb growth rate as immunised CHB lambs had a significantly higher average daily weight gain after challenge than their unvaccinated counterparts. Therefore, the protection of CHB lambs was enhanced by immunisation at weaning, suggesting a synergistic effect when combining vaccination with presumed genetic resistance.

Cellular and humoral immune responses associated with protection in sheep vaccinated against Teladorsagia circumcincta.

Due to increased anthelmintic resistance, complementary methods to drugs are necessary to control gastrointestinal nematodes (GIN). Vaccines are an environmentally-friendly and promising option. In a previous study, a Teladorsagia circumcincta recombinant sub-unit vaccine was administered to two sheep breeds with different levels of resistance against GIN. In the susceptible Canaria Sheep (CS) breed, vaccinates harboured smaller worms with fewer eggs in utero than the control group. Here, we extend this work, by investigating the cellular and humoral immune responses of these two sheep breeds following vaccination and experimental infection with T. circumcincta. In the vaccinated CS group, negative associations between antigen-specific IgA, IgG2 and Globule Leukocytes (GLs) with several parasitological parameters were established as well as a higher CD4+/CD8+ ratio than in control CS animals, suggesting a key role in the protection induced by the vaccine. In the more resistant Canaria Hair Breed (CHB) sheep the vaccine did not significantly impact on the parasitological parameters studied and none of these humoral associations were observed in vaccinated CHB lambs, although CHB had higher proportions of CD4+ and CD8+ T cells within the abomasal lymph nodes, suggesting higher mucosal T cell activation. Each of the component proteins in the vaccine induced an increase in immunoglobulin levels in vaccinated groups of each breed. However, levels of immunoglobulins to only three of the antigens (Tci-MEP-1, Tci-SAA-1, Tci-ASP-1) were negatively correlated with parasitological parameters in the CS breed and they may be, at least partially, responsible for the protective effect of the vaccine in this breed. These data could be useful for improving the current vaccine prototype.

Lessons of the month: A misunderstood teenager with paediatric inflammatory multisystem syndrome - temporarily associated with SARS-CoV-2 admitted under adult medicine.

The emergence of SARS-CoV-2 has proven to be a challenge to healthcare bodies globally. The virus has been associated with a spectrum of clinical features, from anosmia to gastrointestinal upset to multiorgan dysfunction in the most severe cases. Given the range of features observed, it is important to be aware that infectious diseases can present atypically. Furthermore, in many hospitals, including our own, teenagers aged 16 to 18 years old are admitted under the care of adult medical services. Clinicians should be aware of patients presenting with the novel condition of paediatric inflammatory multisystem disorder - temporarily associated with SARS-CoV-2 (PIMS-TS).

Septal Perfoplasty for Management of Symptomatic Nasal Septal Perforation: An Alternative to Surgical Closure.

Importance: Symptomatic septal perforations are often difficult to manage and can have a significant impact on patient quality of life. Available surgical techniques for repair have demonstrated a varying rate of success, presenting a need for reliable interventions targeting symptom control. Objectives: To describe the modified surgical technique here termed septal perfoplasty. To demonstrate that creation of favorable septal perforation characteristics is effective in managing symptoms and improving patient quality of life. Design, Setting, and Participants: A retrospective review of the medical record was performed of patients who underwent the procedure of interest between July 1, 2006 and October 1, 2019 at Vanderbilt University Medical Center. All patients with symptomatic septal perforation who underwent septal perfoplasty within the timeframe reviewed were included. Septal perfoplasty was standardly performed in combination with turbinate reduction in all cases. This was combined with other indicated procedures for chronic sinusitis, repair of vestibular stenosis or nasal deformity. Main Outcomes and Measures: Creation of a well-mucosalized septal perforation, combined with patient-reported acceptable symptom control, was the primary outcome. Secondary outcomes include time to resolution, duration of follow-up, postsurgical complications, and need for further intervention. Results: Twenty patients (70% female; mean [range] age, 45.8 [15-72] years) underwent septal perfoplasty over the course of 13 years. The most common etiology of perforation was trauma (40%), presenting symptom was crusting (95%), and size of perforation repaired was large (60%). Mean follow-up was 37.6 months (range, 1-153 months). Overall, favorable perforation characteristics were created in 95% of cases by the first postoperative appointment. Acceptable symptomatic control was achieved in 18 out of 20 patients (90%), with a median time to improvement of 66 days. Eight patients required additional surgery to address chronic sinusitis or vestibular stenosis. Two patients experienced postoperative infections, treated conservatively with antibiotics. Conclusion and Relevance: Septal perfoplasty is a safe, simple, and effective method for management of symptomatic nasal septal perforation, which provides an alternative to more complicated interventions with comparable rates of symptomatic resolution. This procedure should particularly be considered for patients in which difficult repair is anticipated.

Dengue Fever and COVID-19 Co-Infection at the Emergency Department of a Tertiary Care Hospital in Jeddah, Saudi Arabia.

INTRODUCTION: Co-infection with COVID-19 and other viral or bacterial infections has been described in many studies. Most cases of dengue fever in Saudi Arabia have been reported in Jeddah city and mainly during the summer season. METHOD AND RESULTS: We describe four patients who presented to the emergency department with febrile illness accompanied by symptoms and blood tests suggestive of dengue fever. They also had radiological findings suggestive of COVID-19 chest infection and positive PCR tests. Three of the patients required hospital admission, and all had a good outcome. CONCLUSION: In areas with a high rate of dengue fever, serological screening for dengue fever should be considered as part of COVID-19 investigation, especially if symptoms or a full blood count are suggestive. LEARNING POINTS: Dengue fever (DF) and COVID-19 co-infection has been reported in countries with a high rate of DF.Symptoms and /or full blood count changes suggestive of DF should draw the attention for a DF screening while reviewing patients with suspected COVID-19 infection.

Changes in distribution, morphology and ultrastructure of chloride cell in Atlantic salmon during an AGD infection.

Amoebic gill disease (AGD) is emerging as one of the most significant health challenges affecting farmed Atlantic salmon in the marine environment. It is caused by the amphizoic amoeba Neoparamoeba perurans, with infestation of gills causing severe hyperplastic lesions, compromising overall gill integrity and function. This study used histology, transmission electron microscopy (TEM), immunohistochemistry and transcript expression to relate AGD-associated pathological changes to changes in the morphology and distribution of chloride cells (CCs) in the gills of Atlantic salmon (Salmo salar L.) showing the progression of an AGD infection. A marked reduction in numbers of immunolabelled CCs was detected, and a changing pattern in distribution and morphology was closely linked with the level of basal epithelial hyperplasia in the gill. In addition, acute degenerative ultrastructural changes to CCs at the lesion site were observed with TEM. These findings were supported by the early-onset downregulation of Na+ /K+ -ATPase transcript expression. This study provides supportive evidence that histological AGD lesion assessment was a good qualitative tool for AGD scoring and corresponded well with qPCR genomic Paramoeba perurans quantification. Ultrastructural changes induced in salmon CCs as a result of AGD are reported here for the first time.

Impacts of breed type and vaccination on Teladorsagia circumcincta infection in native sheep in Gran Canaria.

Vaccines and genetic resistance offer potential future alternatives to the exclusive use of anthelmintics to control gastrointestinal nematodes (GIN). Here, a Teladorsagia circumcincta prototype vaccine was administered to two sheep breeds which differ in their relative levels of resistance to infection with GIN. Vaccination of the more susceptible Canaria Sheep (CS) breed induced significant reductions in worm length and numbers of worm eggs in utero (EIU) when compared to control CS sheep. In the more resistant Canaria Hair Breed (CHB), although vaccination induced a reduction in all parasitological parameters analysed, differences between vaccinated and control sheep were not statistically significant. Such interactions between sheep breed and vaccination may allow better integrated control of GIN in future.

A novel, high-welfare methodology for evaluating poultry red mite interventions in vivo.

Optimisation and use of a device for the on-hen in vivo feeding of all hematophagous stages of Dermanyssus gallinae is described. The sealed mesh device contains the mites and is applied to the skin of the hen's thigh where mites can feed on the bird through a mesh which has apertures large enough to allow the mites' mouth-parts to access to the bird but small enough to contain the mites. By optimising the depth and width of the mesh aperture size we have produced a device which will lead to both reduction and refinement in the use of animals in research, allowing the pre-screening of new vaccines and systemic acaricides/insecticides which have been developed for the control of these blood-feeding parasites before progressing to large field trials. For optimal use, the device should be constructed from 105 µm aperture width, 63 µm depth, polyester mesh and the mites (irrespective of life stage) should be conditioned with no access to food for 3 weeks at 4 °C for optimal feeding and post-feeding survival.

Serum and acute phase protein changes in laying hens, infested with poultry red mite.

The poultry red mite (PRM) is one of the most economically important ectoparasites of laying hens globally. This mite can have significant deleterious effects on its fowl host including distress, anemia, reduced egg production, and reduced egg quality. This study was conducted to evaluate the influence of PRM on the serum protein profile in laying hens and its effect on the acute phase proteins (APPs) to assess their potential as biomarkers for mite infestation. Three APPs: alpha-1 acid glycoprotein (AGP), serum amyloid-A (SAA), and ceruloplasmin (CP) were measured in serum samples collected from laying hens at 12 and 17 wk of age, and then for up to 4 mo after a challenge with PRM (starting at 18.5 wk of age). The serum protein profile (SDS-PAGE/nanoflow HPLC electrospray tandem mass spectrometry) and concentration of individual serum proteins (SDS-PAGE-band densitometry) were also compared. Post challenge there was a positive correlation (r = 0.489; P < 0.004) between the levels of SAA and the PRM numbers. The levels of SAA steadily increased after the PRM challenge and were significantly different than the pre-challenge levels at 28, 32, and 36 wk of age (P < 0.01). The PRM numbers also peaked around 31-33 wk of age. The results for AGP and CP in comparison were inconsistent. Proteomics revealed the presence of 2 high molecular weight proteins in the serum between 12 and 17 wk of age. These were identified as Apolipoprotein-B and Vitellogenin-2, and their increase was commensurate with the onset of lay. No other major differences were detected in the protein profiles of blood sera collected pre and post challenge. We conclude that SAA could be used as a useful biomarker to monitor PRM infestation in commercial poultry flocks and that PRM infestation does not disrupt the production of the major proteins in the serum that are associated with egg formation.

Draft Genome Assembly of the Poultry Red Mite, Dermanyssus gallinae.

The poultry red mite, Dermanyssus gallinae, is a major worldwide concern in the egg-laying industry. Here, we report the first draft genome assembly and gene prediction of Dermanyssus gallinae, based on combined PacBio and MinION long-read de novo sequencing. The ∼959-Mb genome is predicted to encode 14,608 protein-coding genes.

Gene silencing by RNA interference in the ectoparasitic mite, Psoroptes ovis.

The presence of components of the RNA interference (RNAi) pathway in Psoroptes ovis, an ectoparasitic mite responsible for psoroptic mange, was investigated through interrogation of the P. ovis genome. Homologues of transcripts representing critical elements for achieving effective RNAi in the mite, Tetranychus urticae and the model organisms Caenorhabditis elegans and Drosophila melanogaster were identified and, following the development of a non-invasive immersion method of double stranded RNA delivery, gene silencing by RNAi was successfully demonstrated in P. ovis. Significant reductions in transcript levels were achieved for three target genes which encode the Group 2 allergen (Pso o 2), mu-class glutathione S-transferase (PoGST-mu1) and beta-tubulin (Poßtub). This is the first demonstration of RNAi in P. ovis and provides a mechanism for mining transcriptomic and genomic datasets for novel control targets against this economically important ectoparasite.

Draft Genome Assembly of the Sheep Scab Mite, Psoroptes ovis.

Sheep scab, caused by infestation with Psoroptes ovis, is highly contagious, results in intense pruritus, and represents a major welfare and economic concern. Here, we report the first draft genome assembly and gene prediction of P. ovis based on PacBio de novo sequencing. The ∼63.2-Mb genome encodes 12,041 protein-coding genes.

Field evaluation of poultry red mite (Dermanyssus gallinae) native and recombinant prototype vaccines.

Vaccination is a desirable emerging strategy to combat poultry red mite (PRM), Dermanyssus gallinae. We performed trials, in laying hens in a commercial-style cage facility, to test the vaccine efficacy of a native preparation of soluble mite extract (SME) and of a recombinant antigen cocktail vaccine containing bacterially-expressed versions of the immunogenic SME proteins Deg-SRP-1, Deg-VIT-1 and Deg-PUF-1. Hens (n=384 per group) were injected with either vaccine or adjuvant only (control group) at 12 and 17 weeks of age and then challenged with PRM 10days later. PRM counts were monitored and, at the termination of the challenge period (17 weeks post challenge), average PRM counts in cages containing birds vaccinated with SME were reduced by 78% (p<0.001), compared with those in the adjuvant-only control group. When the trial was repeated using the recombinant antigen cocktail vaccine, no statistically significant differences in mean PRM numbers were observed in cages containing vaccinated or adjuvant-only immunised birds. The roles of antigen-specific antibody levels and duration in providing vaccine-induced and exposure-related protective immunity are discussed.