Spineless cactus (Opuntia ficus-indica) and saltbush (Atriplex halimus L.) as feed supplements for fattening Awassi male lambs: effect on digestibility, water consumption, blood metabolites, and growth performance

The effect of replacing 13.6% and 20.3% of a total ration of fattening Awassi lambs by two combinations of fresh saltbush (Atriplex halimus) and fresh spineless cactus (Opuntia ficus-indica) cladodes at a ratio of 1.9:1 (TRT1) and 1.7:1 (TRT2) on water intake, digestibility, blood metabolites, and fattening performance was evaluated. Thirty-six lambs with average initial live weight 34.5 ± 4.18 kg were randomly assigned to three diets (control, TRT1, and TRT2). The control received a diet containing 166 g/kg barley straw and 834 g/kg of commercial concentrate mixture; TRT1 comprised 126 g barley straw, 739 g/kg concentrate mixture, 47 g/kg spineless cactus, and 89 g saltbush; TRT2 comprised 67 g/kg barley straw, 704 g/kg commercial concentrate mixture, 86 g/kg spineless cactus, and 144 g saltbush. A growth trial of 100 days (10 days of adaptation and 90 days of collection) followed by a metabolism trial of 17 days (10 days of adaptation and 7 days of a total feces and urine collection) was carried out. Daily dry matter intake, digestibility of crude protein, ether extract and nutrient detergent fiber, nitrogen balance, average daily gain, feed conversion ratio, and blood metabolites were not significantly affected by the treatment. Water consumption in TRT2 was significantly 16% less compared with the control. A combination of saltbush and spineless cactus at a ratio of 1.7:1 (TRT2) replaced 60% of barley straw and 16% of concentrate mixture without adverse effects on health and growth performance of Awassi male lambs. This represents a potential reduction in feed costs for smallholder farmers

Metode Cepat Ekstraksi Dna Corynebacterium Diphtheriae Untuk Pemeriksaan Pcr

Diagnosis of diphtheria caused byCorynebacterium diphtheriaeshould be done immediately since delay of therapy may cause 20-fold increase rate of death. One method of rapid diagnostic to identify diphtheria is by using polymerase chain reaction (PCR). The fundamental issue of this method depends on the DNA, either its quality or quantity. The simple DNA extraction method, which is using mechanical/physical principles with a little of chemical reagents (such as boiling method and the use of sodium hydroxide (NAOH)), will have some benefits, such as easy to be performed, low cost, fast, and environmentally friendly. This study aimed to evaluate effectivity and efficiency of boiling method with NaOH to extract DNA of C. diphtheriae compared to the use of a commercial diagnostic kit for PCR assay. We used C. diphtheriae toxygenic(NCTC 10648) isolates, which are grown in blood agar plates. We then prepared the suspensions of cell/colony in aquadest with several dilutions. Each dilution was extracted using boiling method, NaOH and controlled with the use of a commercial diagnostic kit (QiAmp DNA Minikit). The results were evaluated quantitatively with spectrophotometer and qualitatively with gel electrophoresis. The results showed that the extracted DNA from boiling method with NaOH has an adequate quality and quantity for PCR assay (up to 9 CFU/uL cell/reaction). Therefore, it can be summarized that boiling method with NaOH is effective and efficient to be applied in PCR assay forC. diphtheriae.Key words: boiling extraction method, NaOH, C.diphtheriae, PCRAbstrakKematian kasus difteri yang disebabkan oleh Corynebacterium diphtheriaedapat meningkat 20 kali lipat karena keterlambatan pengobatan sehingga penegakan diagnosis harus dilakukan sesegera mungkin. Salah satu metode diagnostik yang cukup cepat untuk mendeteksi penyakit difteri adalah pemeriksaan polymerase chain reaction(PCR). Keberhasilan pemeriksaan PCR dipengaruhi oleh kualitas dan kuantitas DNA. Metode ekstraksi DNA sederhana menggunakan prinsip mekanik/fisika dengan reagen kimia minimalis, seperti metode pemanasan (boiling)dan penggunaan (NAOH) memiliki beberapa kelebihan, diantaranya mudah dilakukan, biaya murah, waktu yang dibutuhkan singkat dan lebih ramah lingkungan. Penelitian ini bertujuan untuk menilai efektifitas dan efisiensi metode boilingdan NAOH untuk ekstraksi DNA C.diphtheriae dibandingkan dengan penggunaan kit komersial dalam pemeriksaan PCR. Sampel berupa isolat C.diphtheriae toksigenik (NCTC 10648), ditumbuhkan dalam blood agar, kemudian dibuat suspensi sel/koloni dalam aquadest dengan beberapa kali pengenceran. Masing-masing pengenceran diekstraksi dengan metode boilingdan NaOH yang terkontrol dengan kit komersial (QiAmp DNA Minikit). Hasil ekstraksi dinilai secara kuantitatif dengan spektrofotometer dan secara kualitatif dengan gel elektroforesis. Hasil penelitian menunjukkan bahwa DNA hasil ekstraksi dengan metode boilingdan NaOH mempunyai kualitas dan kuantitas yang cukup untuk pemeriksaan PCR hingga 9 CFU/uL sel bakteri/reaksi sehingga dapat disimpulkan bahwa metode boiling dan NaOH cukup efektif dan efisien untuk diaplikasikan pada metode PCR untuk C.diphtheriae.Kata Kunci: boiling, NaOH, C.diphtheriae, PC

Extinction Maps Toward The Milky Way Bulge: Two-Dimensional And Three-Dimensional Tests With APOGEE

Galactic interstellar extinction maps are powerful and necessary tools for Milky Way structure and stellar population analyses, particularly toward the heavily reddened bulge and in the midplane. However, due to the difficulty of obtaining reliable extinction measures and distances for a large number of stars that are independent of these maps, tests of their accuracy and systematics have been limited. Our goal is to assess a variety of photometric stellar extinction estimates, including both two-dimensional and three-dimensional extinction maps, using independent extinction measures based on a large spectroscopic sample of stars toward the Milky Way bulge. We employ stellar atmospheric parameters derived from high-resolution H-band Apache Point Observatory Galactic Evolution Experiment (APOGEE) spectra, combined with theoretical stellar isochrones, to calculate line-of-sight extinction and distances for a sample of more than 2400 giants toward the Milky Way bulge. We compare these extinction values to those predicted by individual near-IR and near+mid-IR stellar colors, two-dimensional bulge extinction maps, and three-dimensional extinction maps. The long baseline, near+mid-IR stellar colors are, on average, the most accurate predictors of the APOGEE extinction estimates, and the two-dimensional and three-dimensional extinction maps derived from different stellar populations along different sightlines show varying degrees of reliability. We present the results of all of the comparisons and discuss reasons for the observed discrepancies. We also demonstrate how the particular stellar atmospheric models adopted can have a strong impact on this type of analysis, and discuss related caveats.NSF Astronomy & Astrophysics Postdoctoral Fellowship AST-1203017Physics Frontier Center/Joint Institute for Nuclear Astrophysics (JINA) PHY 08-22648U.S. National Science FoundationAlfred P. Sloan FoundationParticipating InstitutionsU.S. Department of Energy Office of Science ANR-12-BS05-0015-01Astronom

The role of Wilm’s Tumor1 immunohistochemical marker in surface epithelial ovarian tumors

Background: Wilms’ tumor 1 is a tumor suppressor gene. The gene is located in chromosome 11p13. And its expression was found in many solid tumors (including ovarian tumor) and also expressed in hematologic malignancies, Recent studies found that WT1 to be involved in angiogenesis. Objectives: To evaluate the expression of WT1 in surface epithelial ovarian tumorand study the possibility of using WT1 as replacement of both;ovarian tumor marker CA125 and a endothelial cell phenotypic marker CD34. Patients and methods: This is a study of a retrospective ( cross sectional ) of sixty cases with total abdominal hysterectomy and bilateral salpingo - oopherectomy collected from department of Histopathology – Teaching Laboratories / Medical City Teaching Hospital , as well as Al alwya hospital and Al Habibia hospital in Baghdad during the period of study from December 2007 to December 2012. Thirty cases diagnosed as surface epithelial ovarian tumors and thirty cases of histologically normal ovarian tissue which were included as a control group. Formalin - fixed, paraffin - embedded ovarian tissue blocks from 60 cases were used . Three section of 4 micron for each taken and stained with WT1, CD34, and CA125 immunohistochemical marker on positively charged slides. Results: there were a significant correlation between expression of WT1 and histological types of surface epithelial ovarian tumor with a higher expression in serous tumors among other cancer types (P-value < 0.001).There was a significant positive correlation between the expression of WT1 and CA125 scores ( p-value < 0.001).There was a significant correlation between WT1microvessel density (MVD) expression and CD34- microvessel density (MVD) expression in ovarian tumors (P-value = 0.05).On the other hand, there were no significant correlation of WT1 with the age of cases (P-value = 0.9) and with the grade of ovarian tumors ( P-value = 0.23) . Conclusions: The present study demonstrates high expression of WT1 in both tumor and endothelial cells in surface epithelial ovarian tumors, and it had dual usages in evaluation of both ovarian tumor cells and the vascular density. That was proved by demonstrating a significant correlation between WT1 and CA125 expression, and between WT1-MVD and CD34- MVD . There was no statistically significant association between WT1 expression and different tumor grades. There was significance differences in WT1expression among different histological subtypes of primary ovarian carcinomas, with serous carcinoma as the most frequent type

The impact of sequencing depth on the inferred taxonomic composition and AMR gene content of metagenomic samples

Shotgun metagenomics is increasingly used to characterise microbial communities, particularly for the investigation of antimicrobial resistance (AMR) in different animal and environmental contexts. There are many different approaches for inferring the taxonomic composition and AMR gene content of complex community samples from shotgun metagenomic data, but there has been little work establishing the optimum sequencing depth, data processing and analysis methods for these samples. In this study we used shotgun metagenomics and sequencing of cultured isolates from the same samples to address these issues. We sampled three potential environmental AMR gene reservoirs (pig caeca, river sediment, effluent) and sequenced samples with shotgun metagenomics at high depth (~ 200 million reads per sample). Alongside this, we cultured single-colony isolates of Enterobacteriaceae from the same samples and used hybrid sequencing (short- and long-reads) to create high- quality assemblies for comparison to the metagenomic data. To automate data processing, we developed an open- source software pipeline, ‘ResPipe’

Factors associated with plasmid antibiotic resistance gene carriage revealed using large-scale multivariable analysis

Plasmids are major vectors of bacterial antibiotic resistance, but understanding of factors associated with plasmid antibiotic resistance gene (ARG) carriage is limited. We curated > 14,000 publicly available plasmid genomes and associated metadata. Duplicate and replicate plasmids were excluded; where possible, sample metadata was validated externally (BacDive database). Using Generalised Additive Models (GAMs) we assessed the influence of 12 biotic/abiotic factors (e.g. plasmid genetic factors, isolation source, collection date) on ARG carriage, modelled as a binary outcome. Separate GAMs were built for 10 major ARG types. Multivariable analysis indicated that plasmid ARG carriage patterns across time (collection years), isolation sources (human/livestock) and host bacterial taxa were consistent with antibiotic selection pressure as a driver of plasmid-mediated antibiotic resistance. Only 0.42% livestock plasmids carried carbapenem resistance (compared with 12% human plasmids); conversely, tetracycline resistance was enriched in livestock vs human plasmids, reflecting known prescribing practices. Interpreting results using a timeline of ARG type acquisition (determined by literature review) yielded additional novel insights. More recently acquired ARG types (e.g. colistin and carbapenem) showed increases in plasmid carriage during the date range analysed (1994–2019), potentially reflecting recent onset of selection pressure; they also co-occurred less commonly with ARGs of other types, and virulence genes. Overall, this suggests that following acquisition, plasmid ARGs tend to accumulate under antibiotic selection pressure and co-associate with other adaptive genes (other ARG types, virulence genes), potentially re-enforcing plasmid ARG carriage through co-selection

The ZEPLIN-III dark matter detector: instrument design, manufacture and commissioning

We present details of the technical design and manufacture of the ZEPLIN-III dark matter experiment. ZEPLIN-III is a two-phase xenon detector which measures both the scintillation light and the ionisation charge generated in the liquid by interacting particles and radiation. The instrument design is driven by both the physics requirements and by the technology requirements surrounding the use of liquid xenon. These include considerations of key performance parameters, such as the efficiency of scintillation light collection, restrictions placed on the use of materials to control the inherent radioactivity levels, attainment of high vacuum levels and chemical contamination control. The successful solution has involved a number of novel design and manufacturing features which will be of specific use to future generations of direct dark matter search experiments as they struggle with similar and progressively more demanding requirements.Comment: 25 pages, 19 figures. Submitted to Astropart. Phys. Some figures down sampled to reduce siz

The SDSS-III APOGEE Radial Velocity Survey Of M Dwarfs. I. Description Of The Survey And Science Goals

We are carrying out a large ancillary program with the Sloan Digital Sky Survey, SDSS-III, using the fiber-fed multi-object near-infrared APOGEE spectrograph, to obtain high-resolution H-band spectra of more than 1200 M dwarfs. These observations will be used to measure spectroscopic rotational velocities, radial velocities, physical stellar parameters, and variability of the target stars. Here, we describe the target selection for this survey, as well as results from the first year of scientific observations based on spectra that will be publicly available in the SDSS-III DR 10 data release. As part of this paper we present radial velocities and rotational velocities of over 200 M dwarfs, with a v sin i precision of similar to 2 km s(-1) a measurement floor at v sin i = 4 km s(-1). This survey significantly increases the number of M dwarfs studied for rotational velocities and radial velocity variability (at similar to 100-200 m s(-1)), and will inform and advance the target selection for planned radial velocity and photometric searches for low-mass exoplanets around M dwarfs, such as the Habitable Zone Planet Finder, CARMENES, and TESS. Multiple epochs of radial velocity observations enable us to identify short period binaries, and adaptive optics imaging of a subset of stars enables the detection of possible stellar companions at larger separations. The high-resolution APOGEE spectra, covering the entire H band, provide the opportunity to measure physical stellar parameters such as effective temperatures and metallicities for many of these stars. At the culmination of this survey, we will have obtained multi-epoch spectra and radial velocities for over 1400 stars spanning the spectral range M0-L0, providing the largest set of near-infrared M dwarf spectra at high resolution, and more than doubling the number of known spectroscopic a sin i values for M dwarfs. Furthermore, by modeling telluric lines to correct for small instrumental radial velocity shifts, we hope to achieve a relative velocity precision floor of 50 m s(-1) for bright M dwarfs. With three or more epochs, this precision is adequate to detect substellar companions, including giant planets with short orbital periods, and flag them for higher-cadence followup. We present preliminary, and promising, results of this telluric modeling technique in this paper.Center for Exoplanets and Habitable WorldsPennsylvania State UniversityEberly College of SciencePennsylvania Space Grant ConsortiumNSF AST 1006676, AST 1126413National Science FoundationNational Aeronautics and Space Administration NNX-08AE38A, NNX13AB03GAlfred P. Sloan FoundationU.S. Department of Energy Oce of ScienceUniversity of ArizonaBrazilian Participation GroupBrookhaven National LaboratoryUniversity of CambridgeCarnegie Mellon UniversityUniversity of FloridaFrench Participation GroupGerman Participation GroupHarvard UniversityInstituto de Astrosica de CanariasMichigan State/Notre Dame/JINA Participation GroupJohns Hopkins UniversityLawrence Berkeley National LaboratoryMax Planck Institute for AstrophysicsMax Planck Institute for Extraterrestrial PhysicsNew Mexico State UniversityNew York UniversityOhio State UniversityUniversity of PortsmouthPrinceton UniversitySpanish Participation GroupUniversity of TokyoUniversity of UtahVanderbilt UniversityUniversity of VirginiaUniversity of WashingtonYale UniversityMcDonald Observator

Molecular characterization of extended spectrum cephalosporin resistant Escherichia coli isolated from livestock and in-contact humans in Southeast Nigeria

The rise in antimicrobial resistance (AMR) in bacteria is reducing therapeutic options for livestock and human health, with a paucity of information globally. To fill this gap, a One-Health approach was taken by sampling livestock on farms (n = 52), abattoir (n = 8), and animal markets (n = 10), and in-contact humans in Southeast Nigeria. Extended spectrum cephalosporin (ESC)-resistant (ESC-R) Escherichia coli was selectively cultured from 975 healthy livestock faecal swabs, and hand swabs from in-contact humans. Antimicrobial susceptibility testing (AST) was performed on all ESC-R E. coli. For isolates showing a multi-drug resistance (MDR) phenotype (n = 196), quantitative real-time PCR (qPCR) was performed for confirmation of extended-spectrum β-lactamase (ESBL) and carbapenemase genes. Whole-genome sequencing (WGS) was performed on a subset (n = 157) for detailed molecular characterisation. The results showed ESC-R E. coli was present in 41.2% of samples, with AST results indicating 48.8% of isolates were phenotypically MDR. qPCR confirmed presence of ESBL genes, with bla(CTX-M) present in all but others in a subset [bla(TEM) (62.8%) and bla(SHV) (0.5%)] of isolates; none harboured transferable carbapenemase genes. Multi-locus sequence typing identified 34 Sequence Types (ST) distributed among different sampling levels; ST196 carrying bla(CTX-M-55) was predominant in chickens. Large numbers of single nucleotide polymorphisms (SNPs) in the core genome of isolates, even within the same clade by phylogenetic analysis, indicated high genetic diversity. AMR genotyping indicated the predominant bla(CTX-M) variant was bla(CTX-M-15) (87.9%), although bla(CTX-M-55), bla(CTX-M-64,) and bla(CTX-M-65) were present; it was notable that bla(CTX-M-1), common in livestock, was absent. Other predominant AMR genes included: sul2, qnrS1, strB, bla(TEM-1b), tetA-v2, and dfrA14, with prevalence varying according to host livestock species. A bla(CTX-M-15) harbouring plasmid from livestock isolates in Ebonyi showed high sequence identity to one from river/sewage water in India, indicating this ESBL plasmid to be globally disseminated, being present beyond the river environment. In conclusion, ESC-R E. coli was widespread in livestock and in-contact humans from Southeast Nigeria. WGS data indicated the isolates were genetically highly diverse, probably representing true diversity of wild type E. coli; they were likely to be MDR with several harbouring bla(CTX-M-15.) Surprisingly, human isolates had highest numbers of AMR genes and pigs the least