Search

The continuing mutability of the SARS-CoV-2 virus can result in failures of diagnostic assays. To address this, we describe a generalizable bioinformatics-to-biology pipeline developed for the calibration and quality assurance of inactivated SARS-CoV-2 variant panels provided to Radical Acceleration of Diagnostics programs (RADx)-radical program awardees. A heuristic genetic analysis based on variant-defining mutations demonstrated the lowest genetic variance in the Nucleocapsid protein (Np)-C-terminal domain (CTD) across all SARS-CoV-2 variants. We then employed the Shannon entropy method on (Np) sequences collected from the major variants, verifying the CTD with lower entropy (less prone to mutations) than other Np regions. Polyclonal and monoclonal antibodies were raised against this target CTD antigen and used to develop an Enzyme-linked immunoassay (ELISA) test for SARS-CoV-2. Blinded Viral Quality Assurance (VQA) panels comprised of UV-inactivated SARS-CoV-2 variants (XBB.1.5, BF.7, BA.1, B.1.617.2, and WA1) and distractor respiratory viruses (CoV 229E, CoV OC43, RSV A2, RSV B, IAV H1N1, and IBV) were assembled by the RADx-rad Diagnostics core and tested using the ELISA described here. The assay tested positive for all variants with high sensitivity (limit of detection: 1.72–8.78 ng/mL) and negative for the distractor virus panel. Epitope mapping for the monoclonal antibodies identified a 20 amino acid antigenic peptide on the Np-CTD that an in-silico program also predicted for the highest antigenicity. This work provides a template for a bioinformatics pipeline to select genetic regions with a low propensity for mutation (low Shannon entropy) to develop robust ‘pan-variant’ antigen-based assays for viruses prone to high mutational rates.

ABSTRACT: Given the lack of research regarding the effect of microalgal supplementation in dairy cows on milk mineral concentrations, this study investigated the effect of feeding different protein supplements in dairy cow diets on milk, feces, and blood plasma mineral concentrations, associated milk and blood plasma transfer efficiencies, and apparent digestibility. Lactating Finnish Ayrshire cows (n = 8) were allocated at the start of the trial to 4 diets used in a replicated 4 × 4 Latin square design experiment: (1) control diet (CON), (2) a pelleted rapeseed supplement (RSS; 2,550 g/d), (3) a mixture of rapeseed and Spirulina platensis (RSAL; 1,280 g of RSS + 570 g of S. platensis per day), and (4) S. platensis (ALG; 1,130 g of S. platensis per day). In each of the 4 experimental periods, a 2-wk adaptation to the experimental diets was followed by a 7-d sampling and measurement period. Feed samples were composited per measurement period, milk, and feed samples (4 consecutive days; d 17–20), and blood plasma samples (d 21) were composited for each cow period (n = 32). Data were statistically analyzed using a linear mixed effects model with diet, period within square, square and their interaction as fixed factors, and cow within square as a random factor. Cows fed ALG were not significantly different in their milk or blood plasma mineral concentrations compared with CON, although feeding ALG increased fecal concentrations of macrominerals (Ca and Mg) and trace elements (Co, Cu, Fe, I, Mn, and Zn), and reduced their apparent digestibility, compared with CON. When compared with CON and ALG, milk from cows fed RSAL and RSS had lower milk I concentrations (−69.6 and −102.7 μg/kg of milk, respectively), but total plasma I concentrations were not affected significantly. Feeding S. platensis to dairy cows did not affect mineral concentrations in cows' blood or milk, but care should be taken when rapeseed is fed to avoid reducing milk I concentrations which may in turn reduce consumers' I intake from milk and dairy products.

Abstract Purpose There is a great need to reduce the toxicity of chemotherapy used in the management of pancreatic ductal adenocarcinoma (PDAC). Here we explore if regional pressurized delivery of oxaliplatin can minimize peripheral neuropathy in mice. Methods We used an orthotopic PDAC mouse model and delivered a single dose of oxaliplatin through the portal vein using a pressure-enabled system (pancreatic retrograde venous infusion, PRVI). We analyzed the effects of PRVI on tumor burden and peripheral neuropathy using histopathological and functional assays. Results Tumor weights in mice treated with 2 mg/kg oxaliplatin using PRVI were significantly lower than in mice treated with the same dose systemically. This resulted in reduced peripheral neuropathy signatures in PRVI mice compared to the 20 mg/kg systemic dose required to achieve similar tumor control. Conclusion Regional delivery of highly cytotoxic agents using PRVI can reduce the therapeutic dose of these drugs, thereby lowering toxic side effects.

Abstract Background Research is needed to delineate the relative and combined effects of different antibiotic administration and manure management practices in either amplifying or attenuating the potential for antibiotic resistance to spread. Here, we carried out a comprehensive parallel examination of the effects of small-scale (> 55 °C × 3 days) static and turned composting of manures from dairy and beef cattle collected during standard antibiotic administration (cephapirin/pirlimycin or sulfamethazine/chlortetracycline/tylosin, respectively), versus from untreated cattle, on “resistomes” (total antibiotic resistance genes (ARGs) determined via shotgun metagenomic sequencing), bacterial microbiota, and indicator ARGs enumerated via quantitative polymerase chain reaction. To gain insight into the role of the thermophilic phase, compost was also externally heated to > 55 °C × 15 days. Results Progression of composting with time and succession of the corresponding bacterial microbiota was the overarching driver of the resistome composition (ANOSIM; R = 0.424, p = 0.001, respectively) in all composts at the small-scale. Reduction in relative abundance (16S rRNA gene normalized) of total ARGs in finished compost (day 42) versus day 0 was noted across all conditions (ANOSIM; R = 0.728, p = 0.001), except when externally heated. Sul1, intI1, beta-lactam ARGs, and plasmid-associated genes increased in all finished composts as compared with the initial condition. External heating more effectively reduced certain clinically relevant ARGs (bla OXA, bla CARB), fecal coliforms, and resistome risk scores, which take into account putative pathogen annotations. When manure was collected during antibiotic administration, taxonomic composition of the compost was distinct according to nonmetric multidimensional analysis and tet(W) decayed faster in the dairy manure with antibiotic condition and slower in the beef manure with antibiotic condition. Conclusions This comprehensive, integrated study revealed that composting had a dominant effect on corresponding resistome composition, while little difference was noted as a function of collecting manure during antibiotic administration. Reduction in total ARGs, tet(W), and resistome risk suggested that composting reduced some potential for antibiotic resistance to spread, but the increase and persistence of other indicators of antibiotic resistance were concerning. Results indicate that composting guidelines intended for pathogen reduction do not necessarily provide a comprehensive barrier to ARGs or their mobility prior to land application and additional mitigation measures should be considered. Video Abstract.

Antimicrobial resistance (AMR) is a global health issue affecting humans and livestock. Reduction in antimicrobial use (AMU) and appropriate use of antimicrobials in livestock production systems have been encouraged. Lack of access to qualified veterinarians, policies regulating AMU and knowledge of AMU and AMR have been identified as drivers of inappropriate AMU behavior in developing countries. Hence, para-veterinarians take a lead role in providing veterinary services to livestock farmers in developing countries. Our previous work found Fijian farmers lack knowledge and understanding of AMU and AMR. However, the attitude, knowledge, and behavior of Fijian veterinary professionals toward AMU and AMR is currently unknown. Therefore, this qualitative study used face-to-face, semi-structured interviews to explore and understand Fijian veterinarian and para-veterinarians' attitude, knowledge, and behavior toward AMU and AMR. A sample of at least ten participants was targeted and recruited from the Central and Western divisions of Viti Levu, Fiji. The Theory of Planned Behavior (TPB) informed the development of the semi-structured interview guide. The interviews were audio-recorded and analyzed using reflexive thematic analysis and deductively using the TPB framework. Our analysis generated three key themes: 1) Antimicrobials prescribed and used based on availability and cost rather than clinical need, 2) Para-veterinarians awareness and knowledge of AMR influence treatment decisions, and 3) Limited resources impede effective consultation and veterinary service delivery. This study demonstrated para-veterinarians (not veterinarians) lacked knowledge and understanding of AMU and AMR. The availability and cost of antimicrobials rather than clinical justification drove antimicrobial prescribing amongst the para-veterinarians. Veterinarians did not visit farms to provide veterinary services; therefore, para-veterinarians provided the veterinary services to the livestock farmers. Lack of human resources, antimicrobials, and physical resources incapacitated veterinary service delivery, where services to farmers' were delayed or not provided at all. Terms of reference for veterinary service delivery and para-veterinarian training framework targeting prescribing, dispensing, use of antimicrobials and risks associated with inappropriate AMU are recommended as part of antimicrobial stewardship (AMS) programmes. Allocation of physical and human resources to Fijian veterinary services should be considered part of AMS programmes to improve veterinary service delivery to livestock farmers and optimize the AMU at the country level.

Antimicrobial resistance (AMR) is a global health issue affecting humans and livestock. To mitigate AMR risks, responsible use of antimicrobials in livestock production systems have been advocated. Studies have reported patterns of antimicrobial use (AMU) in livestock production systems; however, there is limited information on the drivers of AMU and AMR. Therefore, this study aimed to explore and understand the attitude and knowledge of Fijian livestock farmers on AMU and AMR. Livestock farmers and managers from the Central and Western divisions of Viti Levu, Fiji were recruited using purposive and snowball sampling methods. Face-to-face one-to-one semi-structured qualitative interviews were conducted. Interview questions were informed by the Theory of Planned Behavior (TPB). Interview transcripts were analyzed inductively using reflexive thematic analysis and deductively using the TPB framework. A total of 19 cattle and poultry farmers took part. Our analysis generated four themes: (1) Uninformed use of antimicrobials and unaware of AMR, (2) Safeguarding livestock and generating income source as primary motivators for using antimicrobials (3) Medicine shortage results in hoarding and self-prescribing, and (4) Farm decisions on AMU and livestock management influenced by foreign farmers and veterinarians. Livestock farmers used medicines in livestock production; however, they could not differentiate amongst different types of medicine, including antimicrobials. Antimicrobials were used to prevent diseases in livestock and promote production of food and financial security but without any awareness of the risks of AMR. Additionally, farmers hoarded and self-prescribed medicines. Farmers rationed antimicrobials by not completing the entire course of antibiotics to save them for future use. Based on past experiences, farmers expressed dissatisfaction with the veterinary services provided by the government. They sought help online and from foreign farmers and veterinarians. We propose the need for antimicrobial stewardship (AMS) programmes focused on promoting rational use of antimicrobials and awareness of AMR amongst farmers in the Fijian livestock production systems. These programmes need to consider the anthropological, socio-cultural, economic, and environmental factors driving AMU. Future studies are underway to explore the attitude and knowledge of Fijian veterinarians, para-veterinarians and pharmacists on AMU and AMR to gain a broader systems knowledge to inform the design of AMS programmes.

The importance of beef production for economy of Brazil and the growing demand for animal protein across the globe warrant an improvement in the beef production system. Although most attention has been on modulation of the rumen microbiome to improve ruminant production, the role of the lower gut microbiome in host health and nutrition remains relatively unexplored. This work aimed to investigate the taxonomy and functional variations in the fecal microbiome of Brazilian beef cattle reared in two different production systems using a metagenomic approach. Sixty male beef cattle from six farms representing semi-intensive (I, n = 2) and traditional (T, n = 4) Brazilian beef production systems were enrolled in the study. Shotgun sequencing was used to characterize taxonomic and functional composition and diversity of the microbiome in fecal samples collected from each animal. Fecal samples were analyzed for copper (Cu), lead (Pb), nitrogen (N), phosphorous (P), selenium (Se), and zinc (Zn) and stable isotopes of carbon (13C) and nitrogen (15N). The fecal microbiome was influenced by the beef production systems with greater functional and lower taxonomic diversity in beef cattle feces from I systems compared with that from T systems. The concentration of N, P, and Zn was higher in beef cattle feces from I systems compared with that from T systems and was associated with taxonomic and functional profile of fecal microbiome in I system, suggesting the role of fecal nutrients in shaping system-specific microbiome. Semi-intensive management practices led to a more complex but less connected fecal microbiome in beef cattle. The microbial community in beef cattle feces from I systems was characterized by greater abundance of beneficial bacteria (phylum Firmicutes and butyrate-producing bacteria family Lachnospiraceae and genera Anaerostipes, Blautia, Butyrivibrio, Eubacterium, Roseburia, and Ruminococcus). In addition, the fecal abundance of microbial genes related to immune system, nutrient metabolism, and energy production was greater in beef cattle raised under I systems compared with that under T systems. Findings of the current study suggest that semi-intensive management practices could facilitate the development of a healthier and more efficient fecal microbiome in beef cattle by driving an increase in the abundance of beneficial bacteria and functional genes.

The importance of neurorehabilitation services for people with disabilities is getting well-recognized in low- and middle-income countries (LMICs) recently. However, accessibility to the same has remained the most significant challenge, in these contexts. This is especially because of the non-availability of trained specialists and the availability of neurorehabilitation centers only in urban cities owned predominantly by private healthcare organizations. In the current COVID-19 pandemic, the members of the Task Force for research at the Indian Federation of Neurorehabilitation (IFNR) reviewed the context for tele-neurorehabilitation (TNR) and have provided the contemporary implications for practicing TNR during COVID-19 for people with neurological disabilities (PWNDs) in LMICs. Neurorehabilitation is a science that is driven by rigorous research-based evidence. The current pandemic implies the need for systematically developed TNR interventions that is evaluated for its feasibility and acceptability and that is informed by available evidence from LMICs. Given the lack of organized systems in place for the provision of neurorehabilitation services in general, there needs to be sufficient budgetary allocations and a sector-wide approach to developing policies and systems for the provision of TNR services for PWNDs. The pandemic situation provides an opportunity to optimize the technological innovations in health and scale up these innovations to meet the growing burden of neurological disability in LMICs. Thus, this immense opportunity must be tapped to build capacity for safe and effective TNR services provision for PWNDs in these settings.

We present supplementary data for the published article, “Hitting the diagnostic sweet spot: Point-of-care SARS-CoV-2 salivary antigen testing with an off-the-shelf glucometer” [1]. The assay described is designed to be performed at home or in a clinic without expensive instrumentation or professional training. SARS-CoV-2 is detected by an aptamer-based assay that targets the Nucleocapsid (N) or Spike (S) antigens. Binding of the N or S protein to their respective aptamer results in the competitive release of a complementary antisense-invertase enzyme complex. The released enzyme then catalyzes the conversion of sucrose to glucose that is measured by an off-the-shelf glucometer. The data presented here describe the optimization of the assay parameters and their contribution to developing this aptamer-based assay to detect SARS-CoV-2. The assay performance was checked in a standard buffer, contrived samples, and patient samples validated with well-established scientific methods. The resulting dataset can be used to further develop glucometer-based assays for diagnosing other communicable and non-communicable diseases.

The recent development and mass administration of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) vaccines allowed for disease control, reducing hospitalizations and mortality. Most of these vaccines target the SARS-CoV-2 Spike (S) protein antigens, culminating with the production of neutralizing antibodies (NAbs) that disrupt the attachment of the virus to ACE2 receptors on the host cells. However, several studies demonstrated that the NAbs typically rise within a few weeks after vaccination but quickly reduce months later. Thus, multiple booster administration is recommended, leading to vaccination hesitancy in many populations. Detecting serum anti-SARS-CoV-2 NAbs can instruct patients and healthcare providers on correct booster strategies. Several in vitro diagnostics kits are available; however, their high cost impairs the mass NAbs diagnostic testing. Recently, we engineered an ACE2 mimetic that interacts with the Receptor Binding Domain (RBD) of the SARS-2 S protein. Here we present the use of this engineered mini-protein (p-deface2 mut) to develop a detection assay to measure NAbs in patient sera using a competitive ELISA assay. Serum samples from twenty-one patients were tested. Nine samples (42.8%) tested positive, and twelve (57.1%) tested negative for neutralizing sera. The data correlated with the result from the standard commercial assay that uses human ACE2 protein. This confirmed that p-deface2 mut could replace human ACE2 in ELISA assays. Using bacterially expressed p-deface2 mut protein is cost-effective and may allow mass SARS-CoV-2 NAbs detection, especially in low-income countries where economical diagnostic testing is crucial. Such information will help providers decide when a booster is required, reducing risks of reinfection and preventing the administration before it is medically necessary.

Methane (CH4) emission from enteric fermentation of ruminant livestock is a source of greenhouse gases (GHG) and has become a significant concern for global warming. Enteric methane emission is also associated with poor feed efficiency. Therefore, research has focused on identifying dietary mitigation strategies to decrease CH4 emissions from ruminants. In recent years, plant-derived bioactive compounds have been investigated for their potential to reduce CH4 emissions from ruminant livestock. The organosulphur compounds of garlic have been observed to decrease CH4 emission and increase propionate concentration in anaerobic fermentations (in vitro) and in the rumen (in vivo). However, the mode of action of CH4 reduction is not completely clear, and the response in vivo is inconsistent. It might be affected by variations in the concentration and effect of individual substances in garlic. The composition of the diet that is being fed to the animal may also contribute to these differences. This review provides a summary of the effect of garlic and its bioactive compounds on CH4 emissions by ruminants. Additionally, this review aims to provide insight into garlic and its bioactive compounds in terms of enteric CH4 mitigation efficacy, consistency in afficacy, possible mode of action, and safety deriving data from both in vivo and in vitro studies.

An integrated understanding of factors influencing the occurrence, distribution, and fate of antibiotic resistance genes (ARGs) in vegetable production systems is needed to inform the design and development of strategies for mitigating the potential for antibiotic resistance propagation in the food chain. The goal of the present study was to holistically track antibiotic resistance and associated microbiomes at three distinct pre-harvest control points in an agroecosystem in order to identify the potential impacts of key agricultural management strategies. Samples were collected over the course of a single growing season (67 days) from field-scale plots amended with various organic and inorganic amendments at agronomic rates. Dairy-derived manure and compost amendment samples (n = 14), soil samples (n = 27), and lettuce samples (n = 12) were analyzed via shotgun metagenomics to assess multiple pre-harvest factors as hypothetical control points that shape lettuce resistomes. Pre-harvest factors of interest included manure collection during/post antibiotic use, manure composting, and soil amended with organic (stockpiled manure/compost) versus chemical fertilizer. Microbial community resistome and taxonomic compositions were unique from amendment to soil to lettuce surface according to dissimilarity analysis. The highest resistome alpha diversity (i.e., unique ARGs, n = 642) was detected in amendment samples prior to soil application, while the composted manure had the lowest total ARG relative abundance (i.e., 16S rRNA gene-normalized). Regardless of amendment type, soils acted as an apparent ecological buffer, i.e., soil resistome and taxonomic profiles returned to background conditions 67 d-post amendment application. Effects of amendment conditions surprisingly re-emerged in lettuce phyllosphere resistomes, with the highest total ARG relative abundances recovered on the surface of lettuce plants grown in organically-fertilized soils (i.e., compost- and manure-amended soils). Co-occurrence analysis identified 55 unique ARGs found both in the soil amendments and on lettuce surfaces. Among these, arnA and pmrF were the most abundant ARGs co-occurring with mobile genetic elements (MGE). Other prominent ARG-MGE co-occurrences throughout this pre-harvest lettuce production chain included: TetM to transposon (Clostridiodies difficile) in the manure amendment and TriC to plasmid (Ralstonia solanacearum) on the lettuce surfaces. This suggests that, even with imposing manure management and post-amendment wait periods in agricultural systems, ARGs originating from manure can still be found on crop surfaces. This study demonstrates a comprehensive approach to identifying key control points for the propagation of ARGs in vegetable production systems, identifying potential ARG-MGE combinations that could inform future surveillance. The findings suggest that additional pre-harvest and potentially post-harvest interventions may be warranted to minimize risk of propagating antibiotic resistance in the food chain.

Antimicrobial stewardship (AMS) programmes in human health and livestock production are vital to tackling antimicrobial resistance (AMR). Data on antimicrobial use (AMU), resistance, and drivers for AMU in livestock are needed to inform AMS efforts. However, such data are limited in Fiji. Therefore, this study aimed to evaluate the association between farmer (socio-economic, demographic) and livestock production and management factors with AMU. Information was collected using purposive and snowball sampling from 236 livestock farmers and managers located in Central and Western divisions, Viti Levu, Fiji. Multinomial logistic regression was used to determine the factors associated with AMU in farms using an aggregated livestock farm model. Farms that raised cattle only for dairy (farm factor) were more likely to use antibiotics and anthelmintics (p = 0.018, OR = 22.97, CI 1.713, 308.075) compared to mixed cattle and poultry farms. Farms that maintained AMU records were more likely to use antibiotics (p = 0.045, OR = 2.65, CI 1.024, 6.877) compared to farms that did not. Other livestock production and management factors had no influence on AMU on the livestock farms. AMU in livestock farms was not influenced by the socio-economic and demographic characteristics of the farmer. There were differences between livestock enterprises regarding their management. The lack of association between management system and AMU could be because there was so much variation in management system, levels of farmer knowledge and awareness of AMU, and in management of farm biosecurity. Future studies exploring farmers’ knowledge and awareness of AMU and livestock management are required to design AMS programmes promoting prudent AMU in all livestock farms locally.

Antimicrobial resistance (AMR) is a major global threat to human and animal health. The use of antimicrobials in the livestock sector is considered to contribute to AMR. Therefore, a reduction in and prudent use of antimicrobials in livestock production systems have been advocated. This cross-sectional survey aimed to investigate the extent of imprudent antimicrobial use (AMU) and to determine whether the AMU practice was affected by either the farming system or species of farmed livestock in the largest island (Viti Levu) of Fiji. A total of 276 livestock enterprises were surveyed and antimicrobials were used on 309 occasions over 90 days. Overall, in 298 of 309 (96%) incidents, antimicrobials were used imprudently, comprising antibiotics, 160 of 170 (94%) and anthelmintics, 138 of 139 (99%). Prudent use of antibiotics was associated with commercial farming systems (X2 = 13, p = 0.001), but no association was observed with anthelmintic use (p > 0.05). Imprudent antibiotic use was associated with dairy (OR = 7.6, CI = 1.41, 41.57, p = 0.018) followed by layer and beef (p > 0.05) compared to broiler enterprises. Imprudent AMU was more common in the backyard and semi-commercial enterprises compared to commercial broiler enterprises. Policies promoting the prudent use of antimicrobials in Fiji should focus on smaller livestock production systems and enterprises.

In this paper, we first consider a multi-objective Portfolio Selection model and then we add another entropy objective function and next we generalized the model. We solve the problems using Neutrosophic optimization technique. The models are illustrated with numerical examples.

Context: Anatomical and visual outcomes of phacoemulsification and 23-gauge (23G) pars plana vitrectomy (PPV) assisted with perfluorocarbon liquid (PFCL) and 27-gauge twin-chandelier endoillumination for primary rhegmatogenous retinal detachment (RRD). Aims: This study aims to determine the anatomical and visual outcomes of phacoemulsification and 23G PPV for primary RRD assisted with 27-gauge (27G) transconjunctival twin-chandelier endoillumination and bimanual vitreous base shaving under PFCL. Settings and Design: A prospective single tertiary center cohort study of 65 consecutive eyes by a single surgeon between August 2013 and June 2014. Subjects and Methods: The primary outcome measure was the primary retinal reattachment rate. The secondary outcome measure was best-corrected visual acuity (BCVA). All patients were reviewed postoperatively up to 4 months. Statistical Analysis Used: Chi-squared, Student's t-test, and one-way ANOVA test were used in this study. Results: At 4 months, overall retinal reattachment was 83.1% with a single operation and 87.7% with a second surgery. After exclusion of the proliferative vitreoretinopathy (PVR) cases, the primary retinal reattachment rate improved to 93.9% and secondary retinal reattachment rate was 98.0%. The mean BCVA improved from logMAR 1.06–0.83 (P = 0.002). PVR (P = 0.0001), silicone oil tamponade (P = 0.002), macula-off RD (P = 0.12), location of RRD (P = 0.09), and delay of more than 7 days for surgery (P = 0.09) were associated with a higher redetachment rate. The main postoperative complications were small bubbles of PFCL in the posterior segment (18.5%) and subretinal PFCL (3.1%). Conclusions: Chandelier endoillumination and PFCL-assisted 23G PPV with phacoemulsification has reattachment rates and visual outcomes similar to the current standard of RRD repair of different complexities. This surgical technique can be considered an alternative method of repairing RRD, with some advantages.

This study investigated the effect of seaweed supplementation in dairy cow diets on milk yield, basic composition, and mineral concentrations. Thirty-seven Icelandic cows were split into three diet treatments: control (CON, no seaweed), low seaweed (LSW, 0.75% concentrate dry matter (DM), 13–40 g/cow/day), and high seaweed (HSW, 1.5% concentrate DM, 26–158 g/cow/day). Cows were fed the same basal diet of grass silage and concentrate for a week, and then were introduced to the assigned experimental diets for 6 weeks. The seaweed mix of 91% Ascophyllum nodosum: 9% Laminaria digitata (DM basis), feed, and milk samples were collected weekly. Data were analyzed using a linear mixed effects model, with diet, week, and their interaction as fixed factors, cow ID as random factor, and the pre-treatment week data as a covariate. When compared with CON milk, LSW and HSW milk had, respectively, less Se (−1.4 and −3.1 μg/kg milk) and more I (+744 and +1649 μg/kg milk), while HSW milk also had less Cu (−11.6 μg/kg milk) and more As (+0.17 μg/kg milk) than CON milk. The minimal changes or concentrations in milk for Se, Cu, and As cannot be associated with any effects on consumer nutrition, but care should be taken when I-rich seaweed is fed to cows to avoid excessive animal I supply and milk I concentrations.

Growth in open-source hardware designs combined with the low-cost of high performance optoelectronic and robotics components has supported a resurgence of in-house custom lab equipment development. We describe a low cost (below $700), open-source, fully customizable high-throughput imaging system for analytical microbiology applications. The system comprises a Raspberry Pi camera mounted on an aluminium extrusion frame with 3D-printed joints controlled by an Arduino microcontroller running open-source Repetier Host Firmware. The camera position is controlled by simple G-code scripts supplied from a Raspberry Pi singleboard computer and allow customized time-lapse imaging of microdevices over a large imaging area. Open-source OctoPrint software allows remote access and control. This simple yet effective design allows high-throughput microbiology testing in multiple formats including formats for bacterial motility, colony growth, microtitre plates and microfluidic devices termed 'lab-on-a-comb' to screen the effects of different culture media components and antibiotics on bacterial growth. The open-source robot design allows customization of the size of the imaging area; the current design has an imaging area of ~420 × 300mm, which allows 29 'lab-on-a-comb' devices to be imaged which is equivalent 3480 individual 1μl samples. The system can also be modified for fluorescence detection using LED and emission filters embedded on the PiCam for more sensitive detection of bacterial growth using fluorescent dyes.

Cell-SELEX is performed to select for cell binding aptamers. We employed an additional selection pressure by using RNAse to remove surface-binding aptamers and select for cell-internalizing aptamers. A common RNA sequence was identified from independent cell-SELEX procedures against two different pancreatic cancer cell lines, indicating a strong selection pressure towards this sequence from the large pool of other available sequences present in the aptamer library. The aptamer is not specific for the pancreatic cancer cell lines, and a similar sequence motif is present in previously published internalizing aptamers. The identified sequence forms a structural motif that binds to a surface protein, which either is highly abundant or has strong affinity for the selected aptamer sequence. Deselecting (removing) this sequence during cell-SELEX may increase the probability of identifying aptamers against cell type-specific targets on the cell surface.

The ability to selectively target disease-related tissues with molecules is critical to the design of effective therapeutic and diagnostic reagents. Recognizing the differences between the in vivo environment and in vitro conditions, we employed an in vivo selection strategy to identify RNA aptamers (targeting motifs) that could localize to tumor in situ. One of the selected molecules is an aptamer that binds to the protein DHX9, an RNA helicase that is known to be upregulated in colorectal cancer. Upon systemic administration, the aptamer preferentially localized to the nucleus of cancer cells in vivo and thus has the potential to be used for targeted delivery.

Ataxia with isolated vitamin E deficiency (AVED) is a rare autosomal recessive condition that is caused by a mutation in the alpha tocopherol transfer protein gene. It is almost indistinguishable clinically from Friedreich’s ataxia but with appropriate treatment its devastating neurological features can be prevented. Patients can present with a progressive cerebellar ataxia, pyramidal spasticity, and evidence of a neuropathy with absent deep tendon reflexes. It is important to screen for this condition on initial evaluation of a young patient presenting with progressive ataxia and it should be considered in patients with a long standing ataxia without any diagnosis in view of the potential therapeutics and genetic counselling. In this case report we present a patient who was initially diagnosed with Friedreich’s ataxia but was later found to have AVED.

Aptamers are a class of therapeutic oligonucleotides that form specific three-dimensional structures that are dictated by their sequences. They are typically generated by an iterative screening process of complex nucleic acid libraries employing a process termed Systemic Evolution of Ligands by Exponential Enrichment (SELEX). SELEX has traditionally been performed using purified proteins, and cell surface receptors may be challenging to purify in their properly folded and modified conformations. Therefore, relatively few aptamers have been generated that bind cell surface receptors. However, improvements in recombinant fusion protein technology have increased the availability of receptor extracellular domains as purified protein targets, and the development of cell-based selection techniques has allowed selection against surface proteins in their native configuration on the cell surface. With cell-based selection, a specific protein target is not always chosen, but selection is performed against a target cell type with the goal of letting the aptamer choose the target. Several studies have demonstrated that aptamers that bind cell surface receptors may have functions other than just blocking receptor-ligand interactions. All cell surface proteins cycle intracellularly to some extent, and many surface receptors are actively internalized in response to ligand binding. Therefore, aptamers that bind cell surface receptors have been exploited for the delivery of a variety of cargoes into cells. This review focuses on recent progress and current challenges in the field of aptamer-mediated delivery.

Although dairy manure is widely applied to land, it is relatively understudied compared to other livestock as a potential source of antibiotic resistance genes (ARGs) to the environment and ultimately to human pathogens. Ceftiofur, the most widely used antibiotic used in U.S. dairy cows, is a 3rd generation cephalosporin, a critically important class of antibiotics to human health. The objective of this study was to evaluate the effect of typical ceftiofur antibiotic treatment on the prevalence of ARGs in the fecal microbiome of dairy cows using a metagenomics approach. β-lactam ARGs were found to be elevated in feces from Holstein cows administered ceftiofur (n = 3) relative to control cows (n = 3). However, total numbers of ARGs across all classes were not measurably affected by ceftiofur treatment, likely because of dominance of unaffected tetracycline ARGs in the metagenomics libraries. Functional analysis via MG-RAST further revealed that ceftiofur treatment resulted in increases in gene sequences associated with "phages, prophages, transposable elements, and plasmids", suggesting that this treatment also enriched the ability to horizontally transfer ARGs. Additional functional shifts were noted with ceftiofur treatment (e.g., increase in genes associated with stress, chemotaxis, and resistance to toxic compounds; decrease in genes associated with metabolism of aromatic compounds and cell division and cell cycle), along with measureable taxonomic shifts (increase in Bacterioidia and decrease in Actinobacteria). This study demonstrates that ceftiofur has a broad, measureable and immediate effect on the cow fecal metagenome. Given the importance of 3rd generation cephalospirins to human medicine, their continued use in dairy cattle should be carefully considered and waste treatment strategies to slow ARG dissemination from dairy cattle manure should be explored.

Cephapirin, a cephalosporin antibiotic, is used by the majority of dairy farms in the US. Fecal and urinary excretion of cephapirin could introduce this compound into the environment when manure is land applied as fertilizer, and may cause development of bacterial resistance to antibiotics critical for human health. The environmental loading of cephapirin by the livestock industry remains un-assessed, largely due to a lack of appropriate analytical methods. Therefore, this study aimed to develop and validate a cephapirin quantification method to capture the temporal pattern of cephapirin excretion in dairy cows following intramammary infusion. The method includes an extraction with phosphate buffer and methanol, solid-phase extraction (SPE) clean-up, and quantification using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The LOQ values of the developed method were 4.02 µg kg(-1) and 0.96 µg L(-1) for feces and urine, respectively. This robust method recovered >60% and >80% cephapirin from spiked blank fecal and urine samples, respectively, with acceptable intra- and inter-day variation (