The Genetic Relationship between Three Trichoderma Species and Inhibitory Effects of T. Harzianum (Rifai) On Ganoderma Boninense

Trichoderma is a genus of soil-borne fungus with abundant reports on its success as biological control agents of a variety of plant pathogens. Antagonistic assessment by dual culture technique showed that 18 out of 48 selected T. harzianum isolates successfully inhibited the mycelial growth of the pathogen Ganoderma boninense (isolate: PER71) at 47.86 to 72.06% with the strongest inhibitor exhibited by strain FA30. Eight samples produced effective volatile antifungal compounds which suppressed the growth of PER71 at 24.528 to 58.70 % over 6 days. When the 10 samples were assayed for the production of non-volatile antifungal compounds, whereby showed the inhibitory effects of 18.35 to 40.16% over 6 days. Strain FA30 was the best inhibitor isolate not only by dual culture inhibition technique, but was also the best producer of volatile and non-voltile inhibitor compounds, at 58.70 and 40.16% respectively. The identifications of species of Trichoderma worldwide are currently deduced from micro-morphological descriptions which is tedious and prone to error. This study undertook a molecular approach, using isozyme electrophoresis, random amplified microsatellite (RAMS) analysis and gene sequence of the internal transcribed spacer-1 (ITS 1) region of the ribosomal DNA of selected Trichoderma isolates. Electrophoretic variation of nine isozyme systems of 47 isolates from 3 species of Trichoderma namely, T. harzianum, T. aureoviride and T. longibrachiatum were studied. The UPGMA cluster analysis of the isozyme data showed the putatively identified T. harzianum to be distinctly separated from the outgroup sample of T. longibrachiatum, whereas T. aureoviride showed a closer genetic relationship to the T. harzianum populations. No distinct conclusion could be drawn from the dendrogram as the level of separation between T. harzianum and T. aureoviride and may not necessarily indicate a difference at the species level. A second molecular approach used was to extract DNA and characterise the sample for their Random Amplified Microsatelite DNA (RAMS) profile. The RAMS generated dendrogram showed that besides the distinct T. longibrachiatum, 2 other lineages were evident by UPGMA analysis. Again the level of taxonomic difference could not be determined. However, no clear separation was obtained by the dendrogram generated by the neighbor-joining (NJ). The third approach was to putatively sequence the samples using the internal transcribed spacer 1 (ITS 1) region of the rDNA. The nucleotide sequences were multiple aligned and compared against the ex-type strains sequences from the NCBI and TrichoBLAST Genbank database. Results showed that 25 out of the 26 putatively identified T. harzianum were in agreement with the genome of the T. harzianum ex-type strain while the single exception belonged to T. virens instead. The 9 putative T. aureoviride were misidentifications where 7 were T. harzianum and 2 were T. virens based on the Genbank database. The single strain of T. longibrachiatum (IMI: 375055) was in agreement with the ex-type strain. This study showed that conventional identification of T. harzianum, despite being done under the best possible care and condition, can still lead to incorrect identifications. Molecular studies by isozyme analysis did not give confident level of separation at the species level. The dendrogram based on UPGMA from RAMS analysis supported the ITS 1 gene sequence analysis but it could not confirm the specific species level. The ITS 1 region study showed that the gene sequences of Trichoderma samples were the most accurate technique for identification, with a bootstrap stability at 100% and a homology of 98-100%

Sensitive determination of Tartrazine (E 102) based on Chitosan/Nanoparticles/MWCNTs Modified Gold Electrode in food and beverage products

Food dyes can be categorized into natural and synthetic color. Tartrazine (E 102) which belong to the family of azo dyes and commonly used in food industry. Tartrazine imparts positive and negative benefits as well, by giving attractive physical appearance and consumer acceptance for over centuries. However, excessively intake of food Tartrazine can cause toxicity and pathogenicity to human. Due to arising of the health issues to mankind, researchers gave attentions for determination of Tartrazine by using analytical and advance methods. Currently, there are several analytical methods available, however, these methods are required skilled persons, time consuming and high cost. Herein, an electrochemical sensor was developed based on the combination of nanomaterials (chitosan, calcium nanoparticles and multiwall carbon nanotubes) for detection of Tartrazine. Electrochemical behavior of the modified gold electrode in the presence of Tartrazine was studied by using cyclic voltammetry and differential pulse voltammetry. Under optimal conditions, the DPV was detected with different concentrations of Tartrazine in the range of 0.1 to 10 ppm, with low detection limit (3.3s/s)

Extraction, analytical and advanced methods for detection of allura red AC (E129) in food and beverages products

Allura Red AC (E129) is an azo dye that widely used in drinks, juices, bakery, meat, and sweets products. High consumption of Allura Red has claimed an adverse effects of human health including allergies, food intolerance, cancer, multiple sclerosis, attention deficit hyperactivity disorder, brain damage, nausea, cardiac disease and asthma due to the reaction of aromatic azo compounds (R = R′ = aromatic). Several countries have banned and strictly controlled the uses of Allura Red in food and beverage products. This review paper is critically summarized on the available analytical and advanced methods for determination of Allura Red and also concisely discussed on the acceptable daily intake, toxicology and extraction methods

Optimization assay of Enzymatic Biosensors for determination of Carbaryl Pesticides

Pesticides are chemicals used worldwide to destroy or control insects, fungi, and other pests. In agriculture, farmers use numerous pesticides to protect seeds and crops. Application of pesticides compounds has indeed significantly increased the yield of agricultural products such as vegetables and fruits. The excessive use of pesticides somehow negatively affects both human and environment. The bioaccumulation characteristic has allowed them to accumulate and remain persistent in the environment for a long period. The presence of pesticides in the environment is particularly hazardous, and prolonged exposure may leads to several health problems like asthma attacks, skin rashes and neurological diseases. Carbaryl is one of the most widely used pesticides due to its powerful effect and low cost. At present, pesticides are detected through conventional analytical techniques. However, such techniques requires high skills personnel, expensive instruments and time-consuming. A demand for simple, fast and effective method is necessary for pesticide detection. This lead to the development of enzymatic biosensor which the objective is to immobilize butyrycholinesterase enzyme based on chitosan onto the glassy carbon electrode via cross-linking with glutaraldehyde. Optimization of the experimental parameters for the biosensor performance was conducted using cyclic voltammetry which includes pH, time, scan rate and the effect of methylene blue. Upon the optimizations, it found that pH7 of electrolyte solution, 40s of response time and 50mVs-1 was identified to provide the optimum conditions for the proposed biosensor that potentially can be used as a tool for pesticide detection. The optimized parameters will be employed for further experiments for designation of sensitive enzymatic biosensor for detection of pesticides from the vegetables

Highly sensitive determination of sunset yellow FCF (E110) in food products based on Chitosan/Nanoparticles/MWCNTs with modified gold electrode

Sunset Yellow belongs to the family of azo dyes, commonly used in food industry. High consumption of Sunset Yellow can cause health problem to human. Due to arising of the health issues, there are several analytical methods available for determination of Sunset Yellow. However, these methods are required skilled manpower, complicated procedures, time consuming and high cost. Herein, an electrochemical sensor was developed based on the combination of chitosan (CHIT), calcium oxide nanoparticles (CaONPs) and multiwall carbon nanotubes (MWCNTs) sensing film for detection of Sunset Yellow in food products. Electrochemical behavior of the modified gold electrode in the presence of Sunset Yellow was studied by using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The morphological characteristics of CHIT/CaONPs/MWCNTs were observed under scanning electron microscope and transmission electron microscope. Under optimal conditions, the DPV was detected with different concentrations of Sunset Yellow in the range of 0.9 to 10 ppm, with detection limit of 0.8 ppm. The developed method has successfully applied for monitoring the presence of Sunset Yellow with different food products including candy, royal jelly, ice cream and soft drink with satisfactory results

Augmented Retting Effect on Kenaf Fibers Using Alkalophilic Pectinase-Producing Bacteria in Combination with Water Solvents

A degumming approach is used in this paper with alkalophilic pectinase-producing bacteria (APPB) and two sources of water solvents to address the existing conventional water retting complexities of kenaf. The incorporation of APPB was confirmed based on their retting feasibilities and multiple cell-wall-degrading enzymatic delicacy. The combinations of APPB with seawater offered retting achievements within six-day retting in non-sterile conditions. These retting niches showed maximum (14.67 U/mL) pectinase activity with fiber separation feasibilities of 4.75 Fried test score. The yielded fiber composition analysis showed a higher cellulose composition (84.65%) and the least amount of hemicellulose, pectin, and ligneous gummy substances. The transmission electron microscopy scan of the yielded fibers showed smooth fiber surfaces, 84.20 µm fiber diameter, and 7.65 g/tex fine fiber compared with uninoculated and combinations of freshwater treatments. The FTIR spectra revealed the cellulosic discrepancies of the retting treatments by monitoring O-H and C=O stretching at ~3300 cm−1 and ~1730 cm−1 wavenumbers. These findings are compelling to yield kenaf fibers of quality considering the existing retting difficulties

Methods for the analysis of Sunset Yellow FCF (E110) in food and beverage products-a review

Food colorants are categorized into natural and synthetic dyes. One of the famous synthetic food dyes is Sunset Yellow FCF (E110) which belongs to the family of azo dyes and widely used in food industry. However, Sunset Yellow has positive and negative effects as well, by giving attractive physical appearance and consumer acceptance. At the same time, it can cause as attention deficit hyperactivity disorder (ADHD), is a group of behavioural symptoms that include inattentiveness, hyperactivity and impulsiveness, cancer and some other health effects with an excess consumption. Due to the arising of the health issues for mankind, researchers should give more priority to develop advance techniques for determination of Sunset Yellow in food and beverage products. The main aim of this review paper is critically discussed on the acceptable daily intake (ADI), toxicology, extraction methods, and analytical and electrochemical sensor methods for determination of Sunset Yellow

Allozyme variations of Trichoderma harzianum and its taxonomic implications

Electrophoretic variation of nine allozyme systems encoded by 14 gene loci were studied on 47 isolates from 3 species of Trichoderma namely, T. harzianum, T. aureoviride and T. longibrachiatum. Polyacrylamide gel electrophoresis was used to investigate the taxonomic circumscription of T. harzianum populations and to evaluate the levels of genetic variations and the population structure. The Level of genetic variations in T. harzianum populations were moderately high (P= 57.10%, A= 0.7857, Ap= 0.60714 and H = 0.1542) compared to E T. aureoviride and T. longibrachiatum. The genetic variation attributable to differences among populations was 7.857%. The mean gene flow among populations was Nm = 1.3351. Genetic identities (I) ranged from 0.9397 to 0.9642 with a mean of 0.94846. Outcrossing rates based on fixation indices average (t) was 0.2334. Nevertheless, the alleles for α-EST-b showed a very low frequency of 0.0400. The polymorphic locus of MDH1 was of the fast allele of T. harzianum, MD1-a, was prevalent in T. aureoviride and T. longibrachiatum populations. Using a UPGMA cluster analysis, T. harzianum and T. longibrachiatum populations were totally separated in these cluster except T. aureoviride populations. T. harzianum presents high levels of genetic diversity compared with other Trichoderma species

Infectious Bronchitis Virus (Gamma coronavirus) in Poultry: Genomic Architecture, Post-Translational Modifications, and Structural Motifs

Infectious bronchitis virus (IBV) is an avian coronavirus (CoV) that belongs to the genus Gamma coronavirus and has been listed as an important disease by the World Organization for Animal Health (WOAH). It causes highly contagious respiratory, reproductive, and renal diseases in commercial poultry farms. Multiple IBV serotypes and genotypes have been identified in many countries and many detected variants do not provide cross-protection against infection, resulting in repeated outbreaks and significant economic losses worldwide. In addition, the high genetic mutations and recombination events in the prominent genomic regions of IBV, particularly in the spike glycoprotein (S) and nucleocapsid (N) proteins, are directly involved in the evolutionary processes of IBV and lead to increased pathogenicity and tissue tropism. The characterization of the different genotypes and the relationship between the structure, function, post-translational modifications (PTMs), and structural motifs will elucidate the mechanisms that promote replication and pathogenicity and affect the host’s immune response during infection. In this review, we discuss the molecular features of various IBV genes and proteins that contribute to the infection process. We also highlight the common PTMs and structural motifs that occur during protein synthesis and are essential components of IBV ecology

Electrochemical DNA biosensor for the detection of Trichoderma harzianum based on ionic liquid/ZnO nanoparticles/chitosan modified electrode using acridine orange (AO) as a redox indicator

An electrochemical DNA biosensor was developed that is based on a gold electrode modified with a nanocomposite membrane made from an ionic liquid, ZnO nanoparticles and chitosan. A single-stranded DNA probe was immobilized on this electrode. Acridine orange was used as the hybridization probe for monitoring the hybridization of the target DNA. The biosensor was capable of detecting target DNA in the concentration range from 1.0 × 10–14 to 1.8 × 10–4 mol L-1, with a detection limit of 1.0 × 10–15 mol L-1. The approach towards constructing a DNA biosensor allows studies on the hybridization even with crude DNA fragments and also to analyze sample obtained from real samples. The results show that the DNA biosensor has the potential for sensitive detection of a specific sequence of the Trichoderma harzianum gene and provides a quick, sensitive and convenient method for the study of microorganisms