Development of duplex real-time PCR for quick detection of cryptosporidiosis in goats.

Cryptosporidium spp. is the most important foodborne and waterborne pathogens and a leading cause of mortality from foodborne and waterborne gastrointestinal diseases. In neonates of domestic animals, it is associated with consistent diarrhea and dehydration. Cryptosporidium infection begins with the ingestion of sporulated oocytes disseminated by carrier animals that consistently contaminate the environment. Many diagnostic tests are available including microscopy and antigen trap-ELISA, but none of the diagnostic tests available currently cannot differentiate between active and passive infection in the host. In the current study, to address this challenge an mRNA-based duplex TaqMan® probe PCR was developed to target the Cryptosporidium oocyst wall protein gene and 18SSU rRNA gene in a single tube that can detect metabolically active cryptosporidial oocysts. The mRNA transcripts are the direct indicator of any actively replicating cell and they will help decipher the active stages of its lifecycle in a host. This diagnostic assay was standardized by computing transcript copy number-based limit of detection (LOD). For COWP and 18SSU rRNA genes, the LOD was 7.08 × 1004 and 5.95 × 1005 , respectively. During active infections, the oocyst wall protein will be active and so its COWP gene transcripts will act as a marker for active infection. While transcripts for 18SSU rRNA are constitutively expressed in cryptosporidial life cycle. This current diagnostic assay will be a quantitative marker that will help assess the active stages of Cryptosporidium infection in neonates. The disease dynamics will help better understand to formulate the control strategies and contain infection among healthy animals.

How have smallholder farmers used digital extension tools? Developer and user voices from Sub-Saharan Africa, South Asia and Southeast Asia.

Digital extension tools (DETs) include phone calls, WhatsApp groups and specialised smartphone applications used for agricultural knowledge brokering. We researched processes through which DETs have (and have not) been used by farmers and other extension actors in low- and middle-income countries. We interviewed 40 DET developers across 21 countries and 101 DET users in Bihar, India. We found DET use is commonly constrained by fifteen pitfalls (unawareness of DET, inaccessible device, inaccessible electricity, inaccessible mobile network, insensitive to digital illiteracy, insensitive to illiteracy, unfamiliar language, slow to access, hard to interpret, unengaging, insensitive to user's knowledge, insensitive to priorities, insensitive to socio-economic constraints, irrelevant to farm, distrust). These pitfalls partially explain why women, less educated and less wealthy farmers often use DETs less, as well as why user-driven DETs (e.g. phone calls and chat apps) are often used more than externally-driven DETs (e.g. specialised smartphone apps). Our second key finding was that users often made - not just found - DETs useful for themselves and others. This suggests the word 'appropriation' conceptualises DET use more accurately and helpfully than the word 'adoption'. Our final key finding was that developers and users advocated almost ubiquitously for involving desired users in DET provision. We synthesise these findings in a one-page framework to help funders and developers facilitate more useable, useful and positively impactful DETs. Overall, we conclude developers increase DET use by recognizing users as fellow developers - either through collaborative design or by designing adaptable DETs that create room for user innovation.

Plant Mediated Silver Nanoparticles and Mode of Action in Cancer Therapy: A Review.

BACKGROUND: Cancer is a widespread fatal disease associated with the abnormal growth of cells in the body. OBJECTIVES: This article represents applications of biologically synthesized silver nanoparticles and their mode of action in cancer therapy. METHODS: Nanomedicines have been proved to be an effective therapy in the treatment of the disease because of a wide range of applications. Due to the small shape and size, these nanoparticles are emerging to be of novel importance. They are abundantly found in various resources with easy extraction. RESULTS: Biologically synthesized silver nanoparticles are safe for humans as well as for the environment. They may replace the harmful therapies like chemotherapy, etc. used in cancer treatment due to the severe side effects associated with it. Sometimes patient may die because of the side effects. CONCLUSION: These green nanoparticles have great potential to treat and diagnose different cancers. This article laid a research opportunity for the diagnosis and treatment of cancer.

Lipidico Injection Protocol for Serial Crystallography Measurements at the Australian Synchrotron.

A facility for performing serial crystallography measurements has been developed at the Australian synchrotron. This facility incorporates a purpose built high viscous injector, Lipidico, as part of the macromolecular crystallography (MX2) beamline to measure large numbers of small crystals at room temperature. The goal of this technique is to enable crystals to be grown/transferred to glass syringes to be used directly in the injector for serial crystallography data collection. The advantages of this injector include the ability to respond rapidly to changes in the flow rate without interruption of the stream. Several limitations for this high viscosity injector (HVI) exist which include a restriction on the allowed sample viscosities to >10 Pa.s. Stream stability can also potentially be an issue depending on the specific properties of the sample. A detailed protocol for how to set up samples and operate the injector for serial crystallography measurements at the Australian synchrotron is presented here. The method demonstrates preparation of the sample, including the transfer of lysozyme crystals into a high viscous media (silicone grease), and the operation of the injector for data collection at MX2.

Antimicrobial and anti-adherence activity of various combinations of coffee-chicory solutions on Streptococcus mutans: An in-vitro study.

CONTEXT: Several studies have demonstrated the activity of natural plants on the dental biofilm and caries development. But few studies on the antimicrobial activity of coffee-based solutions were found in the literature. Further there was no study available to check the antimicrobial effect of coffee solutions with different percentages of chicory in it. AIMS: To evaluate the antimicrobial activity of different combinations of coffee-chicory solutions and their anti-adherence effect on Streptococcus mutans to glass surface. MATERIALS AND METHODS: Test solutions were prepared. For antimicrobial activity testing, tubes containing test solution and culture medium were inoculated with a suspension of S. mutans followed by plating on Brain Heart Infusion (BHI) agar. S. mutans adherence to glass in presence of the different test solutions was also tested. The number of adhered bacteria (CFU/mL) was determined by plating method. STATISTICAL ANALYSIS: Statistical significance was measured using one way ANOVA followed by Tukey's post hoc test. P value < 0.05 was considered statistically significant. RESULTS: Pure chicory had shown significantly less bacterial count compared to all other groups. Groups IV and V had shown significant reduction in bacterial counts over the period of 4 hrs. Regarding anti-adherence effect, group I-IV had shown significantly less adherence of bacteria to glass surface. CONCLUSIONS: Chicory exerted antibacterial effect against S. mutans while coffee reduced significantly the adherence of S. mutans to the glass surface.