Established Laboratory Methods Research Articles

Comparative Sero-Analysis between Copper Levels and the Risk of Acute Myocardial Infarction in District Nowshera

There is an increasing evidence pointing to a possible correlation between AMI and copper levels. Serum copper levels serve as a valuable biomarker for assessing the body's copper status. Objective: To find the changes in serum copper level in patients with Acute Myocardial Infarction (AMI). Methods: This observational study was conducted in District Headquarter Hospital, Nowshera in duration of two years. A total of 240 patients were recruited in this study. Upon admission, demographic information, medical history, and clinical characteristics were collected. Blood samples were obtained for measurement of serum copper levels using established laboratory methods. All data were analyzed through SPSS version 29.0. Results: Among total, 65% males and 35% females and the prevalence of cardiovascular risk factors was notable, with 70% of patients having hypertension, 40% with diabetes mellitus, 60% with dyslipidemia, and 45% identified as smokers. Baseline serum copper levels averaged 110 ± 5.2 µg/dL, within 24 hours of admission, the peak serum copper level significantly increased to 125 ± 18.6 µg/dL. Subsequently, levels decreased gradually over the following days, with mean values of 120 ± 17.1 µg/dL on Day 2, 115 ± 16.4 µg/dL on Day 3, 112 ± 15.9 µg/dL on Day 4, and returning to baseline levels by Day 5, 110 ± 15.2 µg/dL. Conclusions: Serum copper levels exhibit significant changes during the acute phase of AMI, with a peak observed within 24 hours of admission. While patients with anterior infractions showed higher peak copper levels, no significant associations were found between copper levels and traditional cardiovascular risk factors or inflammatory markers

Methodologies for quantitatively comparing the effectiveness of chemical retardants for direct and indirect wildfire suppression using a combustion wind tunnel

Retardant chemicals are often used during wildfire fighting operations where they are primarily dropped from an aircraft onto unburned fuels in a fire's path. While there are established laboratory methods used to determine retarding effectiveness capacity relative to a standard, there are no specific methods for comparing the effectiveness of different retardant formulations in different operational applications. This paper develops four methodologies to assess and compare retardant effectiveness in a variety of tactical uses including retarding durability and effectiveness at different application depths. The resultant assessment methodologies use the consistent controlled environment of a combustion wind tunnel in which conditions reasonably represent field conditions and have low variability enabling fair comparisons between treatments. Evaluations of the assessment methodologies for two commercially available retardants and water are presented and demonstrate low variability in evaluation conditions and consistent, reliable results. These methodologies provide valuable and cost-effective insights into chemical performance and will help wildfire suppression agencies to make informed procurement decisions.

A novel laboratory method for simulating pollinator exposure to agrochemical-laden particulate matter.

Environmental transport and deposition of particulate matter (PM) associated with toxic chemicals has begun to receive attention as a source of risk to pollinators. For example, dust arising from manipulations of insecticide-treated seed has potential to exert toxic effects among non-target insects. Similarly, synthetic steroid growth promoters, antibiotics and multiple insecticides and parasiticides detected in fugitive beef cattle feedyard PM may also negatively impact pollinators since many of these chemicals have been detected on wildflowers and pollinators collected near beef cattle feedyards. Therefore, there is a need to assess risk to pollinators posed by deposition of agrochemical-laden PM, both in the field and the laboratory. Unfortunately, established laboratory methods for simulating PM exposure or toxicity associated with contaminated PM are few and highly situation-specific. Herein we describe development and use of a PM circulation system that can be employed to evaluate toxicity of agrochemical-contaminated PM in the laboratory under controlled conditions. Two model organisms (honeybees (Apis mellifera) and mason bees (Osmia lignaria)) were exposed to agrochemical-free PM in the circulator system, and post-exposure mortality was compared with controls. No significant differences in mortality between exposed and control bees were observed. Next, honeybees and mason bees were exposed to PM spiked with an insecticide known to exert toxic effects to pollinators (thiamethoxam). Bees experienced significantly higher mortality when exposed to thiamethoxam-laden PM at environmentally relevant concentrations as compared to bees exposed to agrochemical-free PM. These results confirm the validity of these methods for use in controlled laboratory PM toxicity tests and offer a source of positive and negative control groups for laboratory and field experiments examining exposure of pollinators to potentially toxic agrochemical-laden PM. This method facilitates generation of more realistic toxicity data than standard contact toxicity tests when pollinator exposure scenarios involve particulate-based agrochemicals or other toxic chemicals.

MagnEtophoretic Slider Assay (MeSA): A simple platform for point-of-care diagnostics

Infectious diseases account for millions of deaths each year. To reduce the number of infectious disease related deaths, diagnostic testing needs to be more accessible to patients in low-income countries as well as developed countries. Current diagnostic methods involve centralized laboratories, trained personnel, and are time-intensive, limiting translation to the point-of-care (POC). Microfluidic devices are a popular alternative for diagnostics due to reduced assay times, reduced sample volume, and lower cost. Microfluidic devices are small (<10 cm) and can perform complex assays. Microfluidic paper-based analytical devices (µPADs) are a popular approach to help translate diagnostics to the POC but historically suffer from poor sensitivity when compared to established laboratory methods. Magnetically labeling analytes allows samples to be sorted resulting in improved sensitivity and specificity. Microfluidic magnetophoresis is the process of manipulating magnetic particles in a magnetic field and offers the ability to wash and concentrate a sample during flow. However, until recently, magnetophoresis has not been used in conjunction with µPADs because magnetophoresis requires complex and expensive instrumentation to control flow. Coupling magnetophoresis with µPADs enables pump-free flow control, simple operation, and low cost. Early magnetophoresis µPADs showed detection limits similar to traditional methods but higher than targets for clinical use. In this work, we demonstrate a novel, simple MagnEtophoretic Slider Assay (MeSA) that is free of any external instrumentation and offers a new platform for POC diagnostics. We demonstrate the assay’s capability through biotin competitive assays and a sandwich immunoassay for E. coli detection. The calculated limit of detection for E. coli was 1.62 × 103 Colony Forming Units per mL (CFU/ml). The work described is a novel and simple microfluidic platform that has potential for a wide range of future applications.

The influence of Trichinella spp. infection on hematological parameters of hens

The purpose of the research is to study dynamics of the peripheral blood cell composition during experimental infection of hens with Trichinella pseudospiralis.Materials and methods. . Two groups were formed according to the principle of analogues, experimental and control, of 10 and 6 laying hens, respectively. The experimental birds were weighed, and the isolate of T. pseudospiralis larvae was administered intragastrically at a dose of 2 sp./g of body weight (4000 thousand larvae per bird). Blood samples were taken from the axillary vein before and at days 4, 7, 11, 14, 18, 28, 35, 43, 53, and 61 after the infection. Hematological parameters (total number of erythrocytes and leukocytes, and hemoglobin concentration) were determined by established laboratory methods with subsequent calculation of the leukogram.Results and discussion. It has been established that trichinellosis in the hens caused by parasitism of non-encapsulated Trichinella spp. species does not affect the number of erythrocytes and hemoglobin in infected birds. Indicators of the Trichinella-associated process are cells in the granulocytic series, eosinophils. The onset of the body's reaction to the infection as expressed by eosinopenia was detected at day 7 after the infection. An increase in the relative content of basophilic and eosinophilic granulocytes was recorded in the hens’ blood from day 18; such parameters reached their maximum values on days 25 and 35, that corresponds to the migratory and muscular stages of the Trichinella-associated process. A relative and absolute quantitative decrease in these groups of cells was subsequently detected in the hens’ blood; however, these values were higher than the control non-infected hens during the entire follow-up (61 days). The dynamics of eosinophils and basophils circulating in the hens’ blood was due to the Trichinella-associated process staging and confirms the predominance of an allergic reaction in the nature of such disease.

Evaluation of the Corrosion Rate at the Bottom of the above – Ground Tank for Storage of Petroleum Products

A study of corrosion in steel fuel storage tanks, which failed after several years of operation highlighted intense pitting and microbiological corrosion. Significantly higher corrosion rates were found than would be expected with normal seawater corrosion. Therefore, the established methodology for predicting the remaining service life of tanks is also questionable. The aim of the study was to evaluate the actual corrosion rates at the bottom of the petroleum product tank. For this purpose, we adapted the established laboratory methods for evaluating the corrosion rate to perform field measurements. The observed corrosion rates in the tank are higher than the currently known values.

Models and methods to characterise levonorgestrel release from intradermally administered contraceptives

Microneedle (MN)-based technologies have been proposed as a means to facilitate minimally invasive sustained delivery of long-acting hormonal contraceptives into the skin. Intradermal administration is a new route of delivery for these contraceptives and therefore no established laboratory methods or experimental models are available to predict dermal drug release and pharmacokinetics from candidate MN formulations. This study evaluates an in vitro release (IVR) medium and a medium supplemented with ex vivo human skin homogenate (SH) as potential laboratory models to investigate the dermal release characteristics of one such hormonal contraceptive that is being tested for MN delivery, levonorgestrel (LNG), and provides details of an accompanying novel two-step liquid–liquid drug extraction procedure and sensitive reversed-phase HPLC–UV assay. The extraction efficiency of LNG was 91.7 ± 3.06% from IVR medium and 84.6 ± 1.6% from the medium supplemented with SH. The HPLC–UV methodology had a limit of quantification of 0.005 µg/mL and linearity between 0.005 and 25 µg/mL. Extraction and detection methods for LNG were exemplified in both models using the well-characterised, commercially available sustained-release implant (Jadelle®). Sustained LNG release from the implant was detected in both media over 28 days. This study reports for the first time the use of biologically relevant release models and a rapid, reliable and sensitive methodology to determine release characteristics of LNG from intradermally administered long-acting drug delivery systems.Graphical abstract

Effect of leaf extracts of Carica papaya (Paw paw) and Pakia biglobosa (African locust bean) on some biochemical parameters of albino rats

In spite of the advances made in orthodox medicine, there has been an increasing interest in herbal medicine. The leaves of Carica papaya and Pakia biglobosa have been reported to contain lots of beneficial medicinal compounds, hence their use in the traditional prevention, management and treatment of ailments/diseases. In this study, the effect of varied concentrations of the ethanol leaf extract of the plants on some biochemical parameters of albino rats was assessed. The phytochemical compositions of the leaves were determined using established standard laboratory methods. Fifty four male Albino rats weighing between 150g-200g were randomly distributed into nine groups of six animals each. A daily single dose of 500mg/kg, 1000mg/kg, 1500mg/kg and 2000mg/kg body weight of either of the extracts was respectively, administered to the eight test groups for fourteen days. The control group was given only feed and water. Biochemical parameters such as the serum activities of Aspartate Aminotransferase, Alanine Aminotranferase and Alkaline phosphatase as well as the serum concentrations of Bilirubin, Albumin, HCO3-, Urea, Creatinine and Na+ were assessed. Result of the analyses showed that the administration of the extracts did not significantly raise the serum activity of ALT. The decreases in serum concentrations of Urea and increase in the concentration of HCO3- were directly proportional to the concentration of the extracts. It can be deduced from this study, that 500mg, 1000mg 1500mg and 2000mg/kg bw of either C. papaya or P. biglobosa did not elicit any marked hepatotoxicological or renotoxicological effect on the experimental animals

Combining Vis–NIR spectroscopy and advanced statistical analysis for estimation of soil chemical properties relevant for forest road construction

AbstractA thorough quantification of soil chemical properties is essential for assessing the engineering properties of forest soils for road design, construction, and maintenance. Here, we investigate the applicability of visible–near‐infrared (Vis–NIR) spectroscopy in conjunction with advanced statistical analysis for estimation of soil chemical properties. Sixty forest soil samples were collected and analyzed for pH, electrical conductivity (EC), CaCO3, organic matter (OM), and cation exchange capacity (CEC) with established laboratory methods. The spectral measurements were performed with a Vis–NIR spectrometer within a range of 350–2,500 nm. To estimate abovementioned soil properties from reflectance spectra, advanced statistical techniques including partial least squares regression (PLSR), hybrid partial least squares and artificial neural networks (PLS–DI–ANN) models, hybrid partial least squares and adaptive neural fuzzy inference system (PLS–DI–ANFIS) models, as well as narrow band spectral indices were applied. The obtained results ​​indicate that the PLS–DI–ANFIS models show great potential for the estimation of pH, EC, OM, and CEC from reflectance spectra and their first derivatives, exhibiting higher R2 values and lower RMSE than the other investigated models. The estimation accuracy for CaCO3, however, was low for all applied methods. The results confirm that Vis–NIR spectroscopy may be applied as a rapid and cost‐efficient alternative to standard chemical soil analysis techniques, aiding forest road design, construction, and maintenance.

Gram positive uropathogens and their antibiogram: Data analysis at a tertiary care hospital in Karnataka

Gram positive uropathogens and their antibiogram: Data analysis at a tertiary care hospital in Karnataka - IJMR- Print ISSN No: - 2394-546X Online ISSN No:- 2394-5478 Article DOI No:- 10.18231/2394-5478.2018.0014, Indian Journal of Microbiology Research-Indian J Microbiol Res

Field analyzers for lead quantification in drinking water samples

Field analyzers for the measurement of lead in drinking water samples are gaining interest from states, water utilities and building managers as rapid, inexpensive and simple tools to quantify lead concentrations. This literature review compares data quality by field analyzers to established laboratory methods and provides practical information (e.g. costs, ease-of-use) on commercial lead analyzers that are based on: (1) Electrochemistry, (2) Colorimetry and (3) Fluorescence. Between and within these three general field analyzer categories, manufacturers specify a variety of protocols to prepare the samples, which differ from the standard acidification in laboratory methods. Review of the literature raised concerns that without adequate sample preparation, field analyzers may not always fully quantify the total lead concentration, including particulate lead, thereby resulting in underestimations. Nonetheless, field analyzers have been used to quickly obtain experimental results in the laboratory, or in the field when access to laboratory equipment was limited, expensive or otherwise impractical. Field analyzers were also successfully used to detect lead from service lines, by a water utility where lead was mostly in the dissolved form. Overall, intrinsic strengths and weaknesses of field analyzers are discussed, to better balance practical convenience and adequate data quality depending on the objective.

A New State of Health Diagnosis Method Utilizing Self-Supervised Deep Learning Via Virtual Tests

For state of the art State of Health (SoH) diagnosis methods a lot of domain-specific expertise and experimental data is needed. Utilizing parameter estimation methods like Kalman filters, the internal resistance and capacity may be estimated. Those methods require a battery model that fits to the battery and the desired application. Established Machine learning methods on the other hand exchange that need of domain knowledge against the need of extensive laboratory experimentation data. This work proposes a new machine learning method for SoH diagnosis that minimizes the amount of required laboratory data while taking no assumptions about the battery chemistry and parameters. The method solely depends on the during operation directly measurable quantities voltage and current with no need of estimating State of Charge. This self-supervised learning approach is enabled by the concept of virtual tests. Virtual tests apply standardized laboratory methods like current pulse tests for internal resistance measurement to a machine learning model. That machine learning model imitates the behavior of the battery by estimating the terminal voltage in dependency of the current, therefore mitigating the need for labeled SoH data. This self-supervised learning method allows an online adaptation of the model to changes of behavior of the aging battery while being used in the working environment. For example for estimating the internal resistance of a traction battery in a battery electric vehicle the current and voltage profiles may be measured while everyday driving.[Figure 1] A machine learning model maps the measured current profile to the voltage profile, therefore memorizing the battery dynamics. Because the model needs to track internal states that represent this dynamic and state of charge a recurrent neural network is a good fit for that application. When the model is trained on the data, a rectangular current profile will be applied to the model, representing a virtual pulse test. The internal resistance can be calculated out of the applied current and the voltage drop of the estimated voltage profile.[Figure 2] In the same way a constant current discharge may be applied for capacity estimation purposes. Later, when the state of health degenerated further, the existing model may be trained on more recent data in order to update the SoH. Figure 3 visualizes the described process. This new method allows the estimation of the SoH with neither specific knowledge about the battery nor elaborate experimental aging data of many instances of the same battery. Instead it utilizes established laboratory methods as is described, for instance, in ISO 12405-4:2018. That way it also does not rely on labeled SoH values for training of the model. Also the accuracy of the method does not suffer from a drift error that increases with the age of the battery. These advantages make it particularly suitable for low quantities and prototypes where extensive laboratory experiments are too expensive or demand too much time. This work was funded by the German Federal Ministry of Education and Research (BMBF, Funding number: 16EMO0262). Figure 1

Comparison of the biofire filmarray meningitis/encephalitis panel to established laboratory methods for the diagnosis of meningoencephalitis

Comparison of the biofire filmarray meningitis/encephalitis panel to established laboratory methods for the diagnosis of meningoencephalitis

A Proposed New Laboratory Protocol for Dispersant Effectiveness Testing Adapted for Subsea Dispersant Injection

The main objective of this study is to develop a new protocol for bench-scale dispersant effectiveness testing adapted for subsea dispersants injection (SSDI). The new approach includes turbulence conditions, dispersant injection techniques and quantification of effectiveness, more representative for a SSDI operation. Results from the new system are compared to dispersant effectiveness measured with established laboratory methods, used for screening dispersants for surface application. The most significant result is that the dispersant ranking obtained with the new test relevant for subsea releases was very different compared to the ranking obtained by screening methods used for surface application of dispersants. This strongly indicates that existing standard methods for dispersant effectiveness testing designed for simulating conditions relevant for surface application of dispersants, are less relevant for SSDI effectiveness testing and justify the need for a more relevant method. Such a Dispersant Injection Effectiveness Test (DIET) is suggested and documented in this study.

Simultaneous determination of aflatoxin B1, fumonisin B1 and deoxynivalenol in beer samples with a label-free monolithically integrated optoelectronic biosensor.

A label-free optical biosensor for the fast simultaneous determination of three mycotoxins, aflatoxin B1 (AFB1), fumonisin B1 (FB1) and deoxynivalenol (DON), in beer samples is presented. The biosensor is based on an array of ten Mach-Zehnder interferometers (MZIs) monolithically integrated along with their respective broad-band silicon light sources onto a single chip. Multi-analyte determination is accomplished by functionalizing the sensing arms of individual MZIs with mycotoxin-protein conjugates. Assay is performed by pumping over the chip mixtures of calibrators or samples with a mixture of specific monoclonal antibodies, followed by reaction with a secondary anti-mouse IgG antibody. Reactions are monitored in real-time by continuously recording the MZI output spectra, which are then subjected to Discrete Fourier Transform to convert spectrum shifts to phase shifts. The detection limits achieved for AFB1, FB1 and DON were 0.8, 5.6 and 24 ng/ml, respectively, while the assay duration was 12 min. Recovery values ranging from 85 to 115% were determined in beer samples spiked with known concentrations of the three mycotoxins. In addition, beers of different types and origin were analysed with the biosensor developed and the results were compared with those provided by established laboratory methods, further supporting the accuracy of the proposed device.

Abstract P4-06-08: An interlaboratory study of complex mutation detection in genes associated with hereditary breast and ovarian cancer highlights both successes and current challenges

Abstract Background: Established laboratory methods, including next-generation sequencing (NGS), can accurately and inexpensively detect certain classes of clinically important mutations in readily accessible protein coding regions of a patient's DNA. These events include single nucleotide variants (SNVs), small insertions/deletions (indels), and sometimes medium sized copy-number deletions or duplications (del/dups). However, other mutation types with clinical relevance are known, and indeed data on over 80,000 patients show that pathogenic, medically important variants of other, more technically challenging types are prevalent in genes related to hereditary breast and ovarian cancer (see associated abstract by Lincoln et al.). We sought to develop and evaluate a reference standard by which laboratories may be able to assess and improve their performance on representative examples of complex, technically “hard” mutations in medically important genes associated with breast, ovarian, and other cancers. Methods: We selected a diverse set of 23 challenging alterations, uncovered by the aforementioned clinical study, all of which are considered pathogenic and potentially actionable under ACMG and NCCN guidelines when uncovered in the germline DNA of a patient. These mutations are in the BRCA1, BRCA2, MLH1, MSH2, MSH6, and PMS2 genes. We generated a single synthetic DNA specimen with all 23 mutations introduced into a known genomic background. Following presentation of this general strategy at a 2016 meeting, 7 laboratories volunteered to collaboratively evaluate this approach. This DNA was created, validated, blinded and provided to these laboratories who sequenced it using a total of 9 different NGS based workflows, including 5 validated clinical tests with customized methods and two sequencing technology vendor (Illumina, Ion Torrent) default pipelines. Multiple target capture biochemical methods were used, as was whole genome sequencing. Results: Twelve of the 23 variants were detected by all 9 laboratory workflows, but just 2 workflows detected all 23. Many, but not all, of these test limitations were previously known. Importantly, evidence of each variant was present in the raw data, suggesting that this strategy is compatible with the diverse biochemical methods used in NGS laboratories today. Raw data for the synthetic variants mimicked that of the endogenous ones (including presenting similar technical artifacts which make these variants “hard”) demonstrating that controls such as these may be useful in the development of methods with improved sensitivity. The vendor-supplied bioinformatics pipelines fared the worst, reinforcing the importance of carefully selecting bioinformatics algorithms and parameters in any laboratory developed test. Discussion: Medically important but technically challenging mutations are prevalent in genes involved in hereditary breast, ovarian, and other cancers. Although patient specimens are also critical, synthetic controls may help efficiently assess the analytic range of any clinical test, highlighting certain strengths and limitations, and can help laboratories develop new methods to improve sensitivity for these challenging variants. Citation Format: Lincoln S, Zook J, Truty R, Chowdhury S, Fellowes A, Mahamdallie S, Ferber M, Cleveland M, Huang C, Tomson F, Klee E, DeSilva W, Seal S, Aradhya S, Nussbaum R, Garlick R, Kingsmore S, Rahman N, Salit M, Shirts B. An interlaboratory study of complex mutation detection in genes associated with hereditary breast and ovarian cancer highlights both successes and current challenges [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P4-06-08.

How Do Advanced Molecular Tests Compare to Routine Clinical Laboratory Evaluation of CSF in Meningoencephalitis? A Study in 10 Urban Emergency Departments Across the USA

BackgroundThe EMERGEncy ID Net Study Group is investigating whether advanced molecular tests (AMT) increase the detection of causative agents in the CSF of patients presenting with meningoencephalitis (ME). We report findings from a pilot study using AMT on 18 CSF samples from 10 US Urban Emergency Departments. The purpose of the pilot was to compare the performance of these four AMT to established clinical laboratory methods.MethodsWe investigated four AMT: (1) BioFire FilmArray ME Panel targeting 14 causative agents; (2) an in-house target-directed next generation sequencing assay targeting 25 agents; (3) a microarray capable of detecting >2,500 agents; and (4) deep metagenomic next generation sequencing. For targeted sequencing, loci from 12 DNA-based and 13 RNA-based pathogens were amplified from the extracts by multiplex PCR. All sequencing was performed on an Illumina MiSeq using 500 cycle v2 Reagent Kits. Reads from the targeted sequencing were aligned to the 25 specific reference target sequences using Bowtie2 while metagenomics reads were processed with the taxonomic sequence classifying software Kraken. For microarray analysis, Lawrence Livermore Microbial Detection Array v2 arrays were hybridized with Cy3-labeled DNA or cDNA. Scanned images of arrays were analyzed by CLiMax 3.1.ResultsEight CSF samples had results positive for well-established causes of ME from prior testing (Table). The pilot study demonstrated none of the four AMT detected all causative agents in the eight CSF samples known to have well-established causes of ME. BioFire and targeted sequencing performed best, both detecting 6/8, metagenomics deep sequencing detected 3/8, and microarray detected 1/8.ConclusionDespite the sophistication of AMT, they cannot detect pathogens they do not target, that are present in small numbers, or that have been eliminated from the CSF by the immune response. Despite the theoretical potential for microarray and metagenomic sequencing to detect thousands of different agents, the agents probably must be present at high levels for detection.Clinical laboratory evaluationBioFire Film ArrayTargeted sequencingMicroarrayMetagenomic sequencing Cryptococcus spp.43302 Streptococcus pneumoniae 11111Enterovirus2220West Nile Virus1N/Aa000 aWest Nile Virus is not on the BioFire Panel.Disclosures All authors: No reported disclosures.

Effects of supplementation of different levels of garlic (Allium sativum) on selected blood profile and immunity of white leghorn chicken

The study was conducted to evaluate the effect of feeding different levels of garlic powder inclusion on selected blood profile and immunity of white leghorn chicken. A total of 180 chickens (156 layers and 24 cocks) were randomly distributed in to 12 pens and assigned to 4 treatments. Treatments were rations containing 0, 1, 2, and 3% garlic powder for T1, T2, T3 and T4, respectively. The CP and ME content of treatment rations were 16-16.6% and 3021.31-3244.4 kcal/kg DM, respectively. Blood profile parameters were determined using established laboratory methods. The value of hemoglobin (Hb) increased insignificantly due to supplementation of different levels of garlic powder. Total white blood cell count (TWBC), basophile, lymphocytes, heterophils and monocytes were not affected (P > 0.05) by treatments. But, slight rise in lymphocyte and heterophil counts were observed in garlic supplemented groups which may be due to immuno-stimulatory effects of garlic. Packed cell volume (PCV) and eosinophils were affected (P < 0.05) by treatments, PCV (38.1, 45,2,41.5 and 39. 2), eosinophils (4.9, 3.2, 3 and 2.8), for T1, T2, T3, and T4, respectively. Mean values of total protein (g/dl) was not affected (P > 0.05) by treatment. The mean values of total immunoglobulin (gm/dl) (3.53, 4.09, 5.58, 3.04) was significantly (P<0.05) higher in T3 compared to other treatments. Generally, inclusion of 2% garlic powder has significantly improved total immunoglobulin. But it significantly lowered eosinophils compared to control group. The present study revealed that mixing layer diets with 1-3% garlic powder could be used in practical layer diets to improve some haematological and immunoglobulin values which might consequently improve blood circulation and immunity of White Leghorns Chickens.

Application of portable XRF to the direct analysis of till samples from various deposit types in Canada

In this study, results by direct portable XRF (‘pXRF’) on unsieved till samples were compared with those by established laboratory methods (aqua regia or fusion ICP-MS and ICP-ES) on the <0.063-mm fraction to determine if the application of direct pXRF in the field would serve as an acceptable guide for immediate follow-up work. Four test sites in Canada were chosen: the Halfmile Lake Cu-Pb-Zn VMS deposit; the intrusion-hosted W-Mo Sisson deposit; a Pb-Zn Mississippi Valley–type (MVT) deposit in the Pine Point district; and the Triple B kimberlite. Unsieved till samples from the GSC archive collection were used for this study and included samples from background areas, immediately overlying, and at various distances down-ice of each deposit. Ziploc® and Whirl-Pak® bags that were used to contain the samples in the field were tested for their properties of X-ray attenuation and contamination. In general, the performance of pXRF in the four test areas was very good where concentrations of elements of interest (indicator or pathfinder elements) were substantially above detection limits by this technique (in the low ppm range for many elements). The following elements, shown to be useful indicator elements (important constituents of the ore/commodity) or pathfinder elements (those associated with the commodity elements) by the established methodology, showed similar patterns by pXRF on the unsieved material: Zn, Cu, Pb, and As at Halfmile Lake; W, Mo, Cu, Zn, Pb, and As at the Sisson deposit; Zn, Pb, and Fe at Pine Point; and Ca, Sr, Cr, and Ni at Triple B. Pathfinder elements whose concentrations were too low for determination by pXRF include: Ag and Sb at Halfmile Lake; Ag and Cd at Sisson; Cd, S, and Se at Pine Point; and Co, Mg, P, U, and Th at Triple B. The high background for Bi by pXRF, equivalent to c . 50 ppm, and its noisy signal precluded its use at Halfmile Lake and Sisson. Elements which tended to show poor precision (three analyses each sample) by pXRF in some samples due to sample heterogeneity include Sn, V, and W. Mercury was erroneously reported for the majority of samples in the low ppm range by pXRF whereas its concentration in fact was in the low ppb range. Several Pb-, Zn- ( c . 1% Pb, Zn) and Fe-rich (up to 16% Fe) samples demonstrated spectral interferences by: Pb on As, Th and Se; Zn on Cu; and Fe on Co. Results for six till samples analysed in Ziploc® and Whirl-Pak® bags showed that Ziploc® absorbs fewer low-energy photons and hence is preferable for determining light elements such as Si, K and Ca. Supplementary material: Four data-sets are available at http://www.geolsoc.org.uk/SUP18897

Identification of common horsetail (Equisetum arvense L.; Equisetaceae) using Thin Layer Chromatography versus DNA barcoding.

The global herbal products market has grown in recent years, making regulation of these products paramount for public healthcare. For instance, the common horsetail (Equisetum arvense L.) is used in numerous herbal products, but it can be adulterated with closely related species, especially E. palustre L. that can produce toxic alkaloids. As morphology-based identification is often difficult or impossible, the identification of processed material can be aided by molecular techniques. In this study, we explore two molecular identification techniques as methods of testing the purity of these products: a Thin Layer Chromatography approach (TLC-test) included in the European Pharmacopoeia and a DNA barcoding approach, used in recent years to identify material in herbal products. We test the potential of these methods for distinguishing and identifying these species using material from herbarium collections and commercial herbal products. We find that both methods can discriminate between the two species and positively identify E. arvense. The TLC-test is more cost- and time-efficient, but DNA barcoding is more powerful in determining the identity of adulterant species. Our study shows that, although DNA barcoding presents certain advantages, other established laboratory methods can perform as well or even better in confirming species’ identity in herbal products.