First Report on Molecular Identification of Moniezia expansa in Sheep from Mannavanur, Palani Hills, Tamil Nadu, India.

PURPOSE: Tape worm infection is common among sheep at SRRC, Mannavanur, Palani hills, Tamil Nadu, India. The aim of the present study is to find out the cestode species infecting the sheep being maintained at SRRC, Mannavanur, by means of molecular method. METHODS: During the second week of June 2021, the hogget flock of sheep (comprising both Bharat Merino and Avikalin sheep breeds) was drenched on empty stomach with commercial preparation of anthelmintic drug containing Niclosamide plus Albendazole, as per the standard dose specified by the manufacturer (Niclozole™: each 5 ml contains 500 mg of Niclosamide and 150 mg of Albendazole: dose for sheep-10 ml/15 kg body weight). The tapeworms expelled in dung by the drug-treated sheep were collected, washed in PBS (pH 7.2), and fixed in between two glass slides using 10% formalin. Furthermore, cytochrome c oxidase subunit I (Cox-I) gene-based PCR was carried out. Only partial sequence (1593 bp) of Cox-I gene of Moniezia expansa from Sheep at SRRC, Mannavanur, Tamil Nadu, India was obtained by PCR. The PCR amplified fragment was cloned into pGEM-T vector and the recombinant plasmid was sequenced. The obtained nucleotide sequences of Cox-I gene of the M. expansa from Indian sheep were analysed with that of 27 more cestode species from different mammalian species (available in GenBank) using bioinformatics tools. RESULTS: The species of the tapeworm was identified as Moniezia species by the Department of Veterinary Parasitology, VC& RI, Orathanadu, TANUVAS by the standard Acidic alum carmine staining method. Due to the ambiguity in the conventional method, Cox-I gene-based PCR and subsequent gene sequencing protocols were used for the identification of the species of cestode infecting sheep at SRRC, Mannavanur, and it was confirmed as M. expansa upon BLAST analysis. Moniezia expansa from SRRC, Mannavanur is having 100% sequence identity at nucleotide level with that of M. expansa from Sengal/Ethiopia. M. benedeni shared 87-88% nucleotide identity with Indian M. expansa. With taenids, the share of percent nucleotide identity of Indian M. expansa ranged from 79 to 81%. M. expansa from Indian sheep was clustering with other anaplocephalids from various mammalian species in the analysis of phylogenetic tree based on Cox-I nucleotide sequences. CONCLUSION: From the present study, it is concluded that M. expansa is the anoplocephalid cestode infecting the sheep at Mannavanur, Tamil Nadu, India. To our knowledge, this is the first report on partial nucleotide sequences of Cox-I gene of M. expansa from Sheep of Indian peninsula. An investigation on the involvement of oribatid mites as the vector in the transmission of M. expansa among sheep at SRRC, Mannavanur needs to be carried out.

Co-thermal liquefaction of Prosopis juliflora biomass with paint sludge for liquid hydrocarbons production.

In this study, Prosopis juliflora biomass was co-liquefied with hydrocarbons rich paint waste for next generation fuel (bio-oil) production. Co-liquefaction (HTL) was performed at varying biomass to paint waste ratios (1:0, 0:1, 1:1, 2:1 and 1:2) at different temperatures from 340 to 440 °C for a holding time of 60 min. Bentonite catalyst was added from 1 to 5% wt. to the HTL reactor. Gas Chromatography-Mass Spectroscopy (GC-MS) and Fourier Transform Infrared Spectroscopy (FTIR) analysis were carried out for bio-oil and HTL aqueous phase. Maximum bio-oil yield was around 49.26% wt. at 420 °C, 2:1 blend and 4% wt. of bentonite catalyst. Energy and carbon recovery of bio-oil was around 70% and 96% respectively. Additionally, HTL aqueous phase was analysed and it showed presence of acids molecules in it. The gas from HTL process contained Carbon dioxide (46.25%), Carbon monoxide (6.38%), Methane (9.35%) and hydrogen (24.53%).

Pharmacological activities of antroquinonol- Mini review.

Antrodia camphorata is an expensive mushroom that grows on the inner cavity of an endangered native tree of Taiwan namely Cinnamomum kanehirai Hayata. It is used as a traditional medicine in Taiwan and has several health benefits including free radical scavenging, anti-inflammatory, antimicrobial, hepatoprotective, neuroprotective, antidiabetic, and free radical-induced DNA damage protecting activities. Antroquinonol is a tetrahydro ubiquinone derivative found predominately in the mycelium of Antrodia camphorata, and is characterized by numerous biological and pharmacological activities. Several studies have revealed potential anticancer effects of antroquinonol in various carcinogenic models. Moreover, a phase II clinical trial is ongoing in the US and Taiwan to treat the lung cancer patients with this active compound. The present review aims at depicting a detailed view of the synthetic procedures of antroquinonol as well as deciphering its potential health benefits with a special emphasis on anticancer properties.

Neuropeptide Arginine Vasotocin Positively Affects Neurosteroidogenesis in the Early Brain of Grouper, Epinephelus coioides.

The neuropeptide arginine vasotocin (AVT) has versatile physiological functions in non-mammalian vertebrates. However, the functional association between AVT and neurosteroidogenesis in the early brain of teleosts remains elusive. We thus studied the developmental expression patterns of the avt gene and their V1 type receptor (avt-rv1 ) at various stages of development [90-150 days after hatching (dah)] in relation to neurosteroidogenesis and oestrogen signalling in the early brain of the orange-spotted grouper (Epinephelus coioides). avt and avt-rv1 mRNAs displayed a significantly increase in expression at 110 dah in the telencephalon and diencephalon. Further, avt mRNAs were localised in three magnocellular neuronal populations of the preoptic area, such as parvocellular, magnocellular and gigantocellular preoptic neurones. Intriguingly, the avt transcripts in those neurones were more abundant in 110 dah compared to other ages. Subsequently, dual fluorescence in situ hybridisation analysis showed that the avt and avt-rv1 genes were highly coexpressed with cyp11a1, hsd3b1, cyp17a1, erα, erß and gpr30, which indicates their potential for functional association. Cyp19a1b-immunoreactive positive fibres were found in close proximity to avt-expressing neurones. Moreover, our results showed that exogenous Avt caused a significant increase in the cellular and gene levels of steroidogenic enzymes and oestrogen receptors (ers), whereas the administration of an Avt-rv1 antagonist caused a decrease in the expression of both steroidogenic enzymes and ers genes in the brain. Furthermore, exogenous oestradiol (E2 ) strongly up-regulated avt mRNAs in the grouper brain. Taken together, the present studies suggest that avt and steroidogenesis may positively work together to increase both E2 biosynthesis and early brain development.

Characterization of GM-CSF-inhibitory factor and Uracil DNA glycosylase encoding genes from camel pseudocowpoxvirus.

The present study describes the PCR amplification of GM-CSF-inhibitory factor (GIF) and Uracil DNA glycosylase (UDG) encoding genes of pseudocowpoxvirus (PCPV) from the Indian Dromedaries (Camelus dromedarius) infected with contagious ecthyma using the primers based on the corresponding gene sequences of human PCPV and reindeer PCPV, respectively. The length of GIF gene of PCPV obtained from camel is 795 bp and due to the addition of one cytosine residue at position 374 and one adenine residue at position 516, the open reading frame (ORF) got altered, resulting in the production of truncated polypeptide. The ORF of UDG encoding gene of camel PCPV is 696 bp encoding a polypeptide of 26.0 kDa. Comparison of amino acid sequence homologies of GIF and UDG of camel PCPV revealed that the camel PCPV is closer to ORFV and PCPV (reference stains of both human and reindeer), respectively.

Comparison of virokine from camel pseudocowpoxvirus (PCPV) with interleukin 10 of the Dromedary camel (Camelus dromedarius).

Cellular interleukin-10 (IL-10) gene from the peripheral blood mononuclear cells of the healthy Dromedary camel (Camelus dromedarius) and viral IL-10 (vIL-10) from the skin scabs of the Dromedary camels infected with contagious ecthyma (a parapoxviral infection in the camels) were amplified by polymerase chain reaction, cloned and characterized. Sequence analysis revealed that the open reading frame (ORF) of dromedarian camel IL-10 is 537 bp in length, encoding 178 amino acid polypeptide while open reading frame of vIL-10 from camel is 561 bp, encoding 187 amino acid polypeptide. The Dromedary camel IL-10 exhibited 62.6% and 68.5% sequence identity at the nucleotide and amino acid level, respectively, with vIL-10 from camel. Sequence analysis also revealed that the Dromedary camel IL-10 shared 99.4% and 98.3% identity at the nucleotide and amino acid level, respectively, with the Bactrian camel (Camelus bactrianus). But vIL-10 from camel shared 84.7% and 83.4% sequence identity at the nucleotide and amino acid level, respectively, with vIL-10 from reindeer (Rangifer tarandus), which is a ruminant species belonging to the order Artiodactyla. The present study was conducted to evaluate the evolutionary origin of the camel parapoxvirus with parapoxviruses of cattle and sheep and the resultant sequence analysis revealed that camel parapoxvirus is closely related to cattle parapoxvirus than sheep parapoxvirus (Orf virus).

Cloning and phylogenetic analysis of Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) from Indian dromedaries (Camelus dromedarius).

The cDNAs of two proinflammatory cytokines viz., IL-6 and TNF-α from dromedarian camels were amplified by PCR using bactrian camel sequences and subsequently cloned for sequence analysis. Relationship based on amino acid revealed that dromedarian camel IL-6 shared 99.5% identity both at nucleotide and amino acid level with bactrian camel IL-6 and in case of TNF-α, the identity of dromedarian camel was 99.4% and 99.1% at nucleotide and amino acid level, respectively with that of bactrian camel. Phylogenetic analysis based on their amino acid sequences indicated the close relationship in these cytokine genes between dromedarian camel and other members of camelids.

Self Replicating Gene Vaccine Carrying P1-2A Gene of FMDV Serotype O and its Effects on the Immune Responses of Cattle.

DNA vaccines are considered as alternatives to live attenuated ones for those diseases like foot-and-mouth disease (FMD) where the production and application of live vaccines have been found unsuccessful. However, stability of DNA and the quantity of antigen expressed are the major limitation with naked DNA vaccines. To address these issues self replicating gene vaccine construct was made for foot-and-mouth disease virus (FMDV) type 'O' and studied. The vector for vaccine construct, designated as pSinCMVVac carried CMV promoter and Poly(A) signal sequences at 5' and 3' end of Sindbis replicase gene respectively. Gene for structural protein precursor (P1-2A) of FMDV serotype 'O' was inserted into pSinCMVVac under subgenomic promoter. 5'UTR (untranslated region) of FMDV was introduced upstream of P1-2A to enhance the level of expression of cloned gene. Functionality of the vaccine construct was confirmed in vitro and in vivo. The self-replicating gene vaccine construct was tested in cattle in comparison with naked DNA vaccine carrying P1-2A and 3CD (pUP3CD). Humoral immune response by ELISA and SNT and cellular response by lymphoproliferation assay using MTT were studied. The default approach of using self replicating gene vaccine in high dose and multiple injection in cattle as followed in our studies might result in immunosuppression as this was observed in our subsequent experiments in guinea pigs. Hence based on dose response studies, vaccine strategy needs to be decided. However, the approach of using Sindbis polymerase gene and UTR in FMDV vaccine is the first report and shows future scope of developing such vaccines.

Processing of multimer FMD virus VP1-2A protein expressed in E. coli into monomers.

Expressions of several genes in bacteria were carried out by independent promoter. However, in case of eukaryotes ribosome skipping and introduction of IRES are employed as alternative to multiple translation initiation. Foot and mouth disease virus (FMDV) 2A peptide has been widely used for co-expression of multiple genes in eukaryotic, plant and mammalian systems. The 18 amino acid 2A peptide of FMDV facilitates efficient co-translational dissociation of the polyprotein into discrete protein products. To study the role of 2A in multimeric protein production a construct consisting of tandem repeat of 4 units of C- terminal VP1 linked through 2A sequence was made and expressed in E. coli. Along with tetramer protein, trimer, dimer and monomer proteins were produced. Stability studies showed that the tetramer protein was cleaved to smaller monomer on storage. The results provide scope for using FMDV 2A for expressing multiple genes under a single promoter in prokaryotes.

The use of tyre pyrolysis oil in diesel engines.

Tests have been carried out to evaluate the performance, emission, and combustion characteristics of a single cylinder direct injection diesel engine fueled with 10%, 30%, and 50% of tyre pyrolysis oil (TPO) blended with diesel fuel (DF). The TPO was derived from waste automobile tyres through vacuum pyrolysis. The combustion parameters such as heat release rate, cylinder peak pressure, and maximum rate of pressure rise also analysed. Results showed that the brake thermal efficiency of the engine fueled with TPO-DF blends increased with an increase in blend concentration and reduction of DF concentration. NO(x), HC, CO, and smoke emissions were found to be higher at higher loads due to the high aromatic content and longer ignition delay. The cylinder peak pressure increased from 71 bars to 74 bars. The ignition delays were longer than with DF. It is concluded that it is possible to use tyre pyrolysis oil in diesel engines as an alternate fuel in the future.

Subcutaneous rupture of hepatic hydatid cyst.

Hepatic hydatid cysts are common entities. We present an unusual case of a hepatic hydatid cyst which had ruptured into the subcutaneous tissues of the anterior abdominal wall. The possible causes, imaging features and recent literature are reviewed.