Fusarium oxysporum; its enhanced entomopathogenic activity with acidic silver nanoparticles against Rhipicephalus microplus ticks.

Fusarium oxysporum is an entomopathogenic fungus, and it has anti-biological activity against arthropods. Ticks are blood sucking arthropods which are responsible for transmitting different diseases in humans and animals. The use of chemical insecticides against ticks is not eco-friendly option and results in the development of acaricide resistance. Previously, we had cultured a local isolate of Fusarium oxysporum from soil samples which were identified through microscopy and confirmed through molecular technique. In our previous experiments, we have prepared Silver nanoparticles (AgNP) at pH 7 and they had been characterized through X-Ray Diffraction (XRD), UV-visible and zeta-potential. In our current study, the AgNP were prepared at different pH conditions and characterized through Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The protein molecules of F. oxysporum were charged with Ag ions. F. oxysporum NP were observed to enhance anti-biological activity by killing Rhipicephalus microplus and they caused 100% mortality at pH 4 and pH 5 in 24 h in anti-tick biological assay. Our study is the first report to do biological assay against Rhipicehalus ticks by using Fusarium AgNP at acidic pH. Biological control using entomopathogenic fungi can be the best alternative of the chemical method to control the tick population.

Detection of Yersinia pestis by pesticin fluorogenic probe-coupled PCR.

The <> assay (PE Applied Biosystems) combines PCR with concomitant release of fluorogenic nucleotides for immediate product detection by fluorometry. Yersinia pestis, the etiological agent of bubonic plague, expresses species-specific genes known to be located on two unique plasmids (9.6-kb pPCP and 100.9-kb pMT). Pesticin (pst) is a unique gene located on pPCP which encodes for a bacteriocin. Using fluorogenic probe coupled PCR as few as three copies of pst targets were detected from total Y. pestis genomic DNA. The pst probe used in this report was positive only for pesticinogenic isolates and did not show complementarity with Yersiniae nor with other bacteria targeted in this study suggesting, that the pst probe is very specific for Y. pestis. Under optimal conditions of Mg(2+)concentration and thermal cycle number, addition of extraneous DNA to respective assay mixtures had no effect on detection.

A review of molecular recognition technologies for detection of biological threat agents.

The present review summarizes the state of the art in molecular recognition of biowarfare agents and other pathogens and emphasizes the advantages of using particular types of reagents for a given target (e.g. detection of bacteria using antibodies versus nucleic acid probes). It is difficult to draw firm conclusions as to type of biorecognition molecule to use for a given analyte. However, the detection method and reagents are generally target-driven and the user must decide on what level (genetic versus phenotypic) the detection should be performed. In general, nucleic acid-based detection is more specific and sensitive than immunological-based detection, while the latter is faster and more robust. This review also points out the challenges faced by military and civilian defense components in the rapid and accurate detection and identification of harmful agents in the field. Although new and improved sensors will continue to be developed, the more crucial need in any biosensor may be the molecular recognition component (e.g. antibody, aptamer, enzyme, nucleic acid, receptor, etc.). Improvements in the affinity, specificity and mass production of the molecular recognition components may ultimately dictate the success or failure of detection technologies in both a technical and commercial sense. Achieving the ultimate goal of giving the individual soldier on the battlefield or civilian responders to an urban biological attack or epidemic, a miniature, sensitive and accurate biosensor may depend as much on molecular biology and molecular engineering as on hardware engineering. Fortunately, as this review illustrates, a great deal of scientific attention has and is currently being given to the area of molecular recognition components. Highly sensitive and specific detection of pathogenic bacteria and viruses has increased with the proliferation of nucleic acid and immuno-based detection technologies. If recent scientific progress is a fair indicator, the future promises remarkable new developments in molecular recognition elements for use in biosensors with a vast array of applications.

Detection of Yersinia pestis using branched DNA.

In contrast to target amplification methods, e.g. polymerase chain reaction, the branched DNA (bDNA) signal amplification method quantitates target nucleic acid at physiological levels, involving a series of hybridization reactions without thermal cycling. In this report, we describe a modification of the bDNA assay in which a <> preamplifier oligonucleotide (206 mer) is used in concert with ELISA and light addressable potentiometric sensor (LAPS) formats to detect the plasminogen activator (pla) gene of Yersinia pestis, the etiological agent of plague. Pla is encoded by a 9.6-kb plasmid pPCP, which is essential for virulence. The detection limit of the bDNA-ELISA and LAPS assays is less than 10 000 and 1000 molecules of Y. pestis plasmid DNA, respectively.