Experimental investigation of homogeneous charge compression ignition combustion of biodiesel fuel with external mixture formation in a CI engine.

In parallel to the interest in renewable fuels, there has also been increased interest in homogeneous charge compression ignition (HCCI) combustion. HCCI engines are being actively developed because they have the potential to be highly efficient and to produce low emissions. Even though HCCI has been researched extensively, few challenges still exist. These include controlling the combustion at higher loads and the formation of a homogeneous mixture. To obtain better homogeneity, in the present investigation external mixture formation method was adopted, in which the fuel vaporiser was used to achieve excellent HCCI combustion in a single cylinder air-cooled direct injection diesel engine. In continuation of our previous works, in the current study a vaporised jatropha methyl ester (JME) was mixed with air to form a homogeneous mixture and inducted into the cylinder during the intake stroke to analyze the combustion, emission and performance characteristics. To control the early ignition of JME vapor-air mixture, cooled (30 °C) Exhaust gas recirculation (EGR) technique was adopted. The experimental result shows 81% reduction in NOx and 72% reduction in smoke emission.

First report on molecular characterization of Oestrus ovis in sheep from India.

The objective of the present study is to characterize the dipteran larvae species infesting the sheep being maintained at SRRC, Mannavanur, by means of COI gene based PCR. During the last week of May 2021, post mortem examination of the skull of an Avikalin male sheep (20 months old) revealed the presence of larvae in its nasal sinuses. The larvae were washed in PBS (pH 7.2) and preserved in 70% alcohol. Total genomic DNA was isolated from the larvae using an initial step of grinding with liquid Nitrogen in a sterile mortar and pestle. Using the isolated genomic DNA from the larvae as a template, Cytochrome c oxidase subunit I (COI) gene based PCR was employed using the primers designed based on the COI gene of reference isolate of Oestrus ovis available in the GenBank. Full length COI gene (1534 bp) gene of Oestrus ovis in sheep from South India was targeted in the PCR experiment. The pTZ57R/T vector was used for the cloning of the PCR amplified fragment and the confirmed recombinant plasmid was subjected to sequencing experiments. In addition to morphological examination, based on COI gene based PCR, eventual sequencing experiments and BLAST analysis, it was confirmed that the larvae in the nasal sinuses of sheep from South India were Oestrus ovis. The South Indian isolate of Oestrus ovis is sharing 100% sequence identity both at nucleotide and amino acid levels with that of O. ovis from Spain. The North Indian isolate of O. ovis (from Jammu) exhibited 92% and 99% identity at respective nucleotide and amino acid levels with South Indian isolate. With other members of the subfamily Oestrinae, the share of per cent nucleotide and amino acid identities of South Indian O. ovis ranged from 85-86% to 95-96%, respectively. O. ovis from South India was grouped with the other members of Oestrinae from different geographical areas of the globe in the analysis of phylogenetic tree based on COI amino acid sequences. Based on the research findings, it is concluded that Oestrus ovis is the dipteran species infesting the sheep at Mannavanur, Tamil Nadu, India. To our knowledge, this is the first report on full length nucleotide sequences of COI gene of O. ovis in sheep from Indian subcontinent. Supplementary Information: The online version contains supplementary material available at 10.1007/s12639-024-01666-2.

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).

Empirical evidence of effects of stringency amid Covid-19 pandemic spread.

The objective of this paper is to provide an insight on effect of stringency in Covid-19 spread in India especially in Chennai, a city were more lockdown, and restrictions was imposed to control the infection. Even though the restriction was imposed in the country by the end of March 2020, the growth reduction was seen in the mid of June as the awareness was increased. The average Covid-19 case growth was got reduce from 3.43 to 2.62% by July mid. To analysis the impact of stringency, a detailed analysis was done on Chennai city which was imposed with more repeated lockdowns to flatten the curve. We tried to fit a regression line with three difference scenario of data. The results show a promising R-squared and p value, with a right skewed distribution normal probability plot. The impact of lockdown in people's lives in different sectors were also discussed in this paper.

Sindbis virus replicase-based DNA vaccine construct encoding FMDV-specific multivalent epitope gene: studies on its immune responses in guinea pigs.

Foot-and-mouth disease (FMD) is still a perennial global menace affecting livestock health and production. It is imperative to figure out new ways to curb this disease. In this study, a sindbis virus replicase-based DNA vaccine, pSinCMV-Vac-MEG990, encoding a multivalent epitope gene (representing tandemly linked VP1 C-terminal halves of three foot-and-mouth disease virus (FMDV) serotypes) was constructed. In vitro transfection studies in BHK-21 cells revealed that the construct was able to express FMDV-specific antigen but does not overproduce the antigen. Immunization of guinea pigs with the construct at dose rate of 10, 5, 2 and 1 µg per animal through intramuscular route showed significant neutralizing antibody induction at all doses against all serotype tested as compared to non-immunized controls. On viral challenge of guinea pigs 4 week post-immunization with 1000 GPID(50) of FMDV serotype A, it was observed that the immunization not only delayed the appearance and reduced the severity of FMD lesions significantly (P < 0.05) but also provided complete protection in several guinea pigs. In fact, two of six and one of six guinea pigs were completely protected in 10 and 5 µg immunized groups, respectively. These results suggest that the development of the replicase-based DNA vaccine may provide a promising approach as an alternative vaccine strategy for controlling FMD.

A Statistical Non-Parametric data analysis for COVID-19 incidence data.

BACKGROUND: The impact of COVID-19 on the Global scale is tremendously drastic. There are several types of research going on across the world simultaneously to understand and overcome this dire pandemic outbreak. This paper is purely a statistical study on a distinct set of datasets regarding COVID-19 in India. The motivation of this study is to provide an insight into the rapid growth of confirmed COVID-19 cases in India. METHODS: The rapid growth of COVID-19 cases in India started in March 2020. The main objective of this paper is to provide a solid statistical model for the policymaker to handle this kind of pandemic situation in the near future with nonlinear data. In this paper, the data was got from 1st April to 29th November 2020. To come up with a solid statistical model, various nonlinear data such as confirmed COVID-19 cases, maximum temperature, minimum temperature, the total population (state-wise), the total area in km2 (state-wise), and the total rural and urban population count (state-wise) have been analyzed. In this paper, six different Generalized Additive Models (GAM) was identified after a thorough analysis of other researchers' (Xie and Zhu, 2020; Prata et al., 2020) findings. RESULTS: In all perspectives, the results were identified and analyzed. The GAM model regarding total COVID-19 confirmed cases, total population, and the total rural population provides the best average fit of R2 value of 0.934. As the population value is quite high, the author has concise it using logarithm to provide the best p-value of 0.000542 and 0.001407 for a relation between the total number of COVID-19 cases regarding the total population and total rural population respectively.

A Novel Edge-Based Trust Management System for the Smart City Environment Using Eigenvector Analysis.

The proposed Edge-based Trust Management System (E-TMS) uses an Eigenvector-based approach for eliminating the security threats present in the Internet of Things (IoT) enabled smart city environment. In most existing trust management systems, the trust aggregation process completely depends on the direct trust ratings obtained from both legitimate and malicious neighboring IoT devices. E-TMS possesses an edge-assisted two-level trust computation approach for ensuring the malicious free trust evaluation of IoT devices. The E-TMS aims at removing the false contribution on aggregated trust data. It utilizes the properties of the Eigenvector for identifying compromised IoT devices. The Eigenvector Analysis also helps to avoid false detection. The analysis involves a comparison of all the contributed trust data about every single connected device. A spectral matrix will be generated corresponding to the contributions and the received trust will be scaled based on the obtained spectral values. The absolute sum of obtained values will contain only true contributions. The accurate identification of false data will remove the effect of malicious contributions from the final trust value of a connected IoT device. Since the final trust value calculated by the edge node contains only the trustworthy data, the prediction about the malicious nodes will be accurate. Eventually, the performance of E-TMS has been validated. Throughput and network resilience are higher than the existing system.

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.

Molecular characterization of Orf virus isolates from Kodai hills, Tamil Nadu, India.

AIM: The present study was carried out to find out the causative agent of exanthematous skin lesions in sheep maintained by Southern Regional Research Centre, Mannavanur, Kodai hills, Tamil Nadu. MATERIALS AND METHODS: Polymerase chain reaction (PCR) with Orf virus (ORFV) B2L gene-specific primers was carried out by employing the total genomic DNA isolated from the scabs as the template. The ORFV isolates from Kodai hills were characterized by the use of bioinformatics tools. RESULTS: The amino acid identity of ORFV isolate 1 from Kodai hills is having 98.14%, 96.29%, and 83.59% identity with reference strains of ORFV, Pseudocowpox virus, and bovine papular stomatitis virus, respectively. Phylogenetic analysis revealed that ORFV isolates from Kodai hills clustered with the other ORFV isolates from different geographical areas of India. CONCLUSION: The etiological agent of exanthematous skin lesion among sheep of Kodai hills is ORFV.

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.

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.

Anatomical and functional implications of corticotrophin-releasing hormone neurones in a septal nucleus of the avian brain: an emphasis on glial-neuronal interaction via V1a receptors in vitro.

Previously, we showed that corticotrophin-releasing hormone immunoreactive (CRH-IR) neurones in a septal structure are associated with stress and the hypothalamic-pituitary-adrenal axis in birds. In the present study, we focused upon CRH-IR neurones located within the septal structure called the nucleus of the hippocampal commissure (NHpC). Immunocytochemical and gene expression analyses were used to identify the anatomical and functional characteristics of cells within the NHpC. A comparative morphometry analysis showed that CRH-IR neurones in the NHpC were significantly larger than CRH-IR parvocellular neurones in the paraventricular nucleus of the hypothalamus (PVN) and lateral bed nucleus of the stria terminalis. Furthermore, these large neurones in the NHpC usually have more than two processes, showing characteristics of multipolar neurones. Utilisation of an organotypic slice culture method enabled testing of how CRH-IR neurones could be regulated within the NHpC. Similar to the PVN, CRH mRNA levels in the NHpC were increased following forskolin treatment. However, dexamethasone decreased forskolin-induced CRH gene expression only in the PVN and not in the NHpC, indicating differential inhibitory mechanisms in the PVN and the NHpC of the avian brain. Moreover, immunocytochemical evidence also showed that CRH-IR neurones reside in the NHpC along with the vasotocinergic system, comprising arginine vasotocin (AVT) nerve terminals and immunoreactive vasotocin V1a receptors (V1aR) in glia. Hence, we hypothesised that AVT acts as a neuromodulator within the NHpC to modulate activity of CRH neurones via glial V1aR. Gene expression analysis of cultured slices revealed that AVT treatment increased CRH mRNA levels, whereas a combination of AVT and a V1aR antagonist treatment decreased CRH mRNA expression. Furthermore, an attempt to identify an intercellular mechanism in glial-neuronal communication in the NHpC revealed that brain-derived neurotrophic factor (BDNF) and its receptor (TrkB) could be involved in the signalling mechanism. Immunocytochemical results further showed that both BDNF and TrkB receptors were found in glia of the NHpC. Interestingly, in cultured brain slices containing the NHpC, the use of a selective TrkB antagonist decreased the AVT-induced increase in CRH gene expression levels. The results from the present study collectively suggest that CRH neuronal activity is modulated by AVT via V1aR involving BDNF and TrkB glia in the NHpC.

Genetic trend for growth and wool performance in a closed flock of Bharat Merino sheep at sub temperate region of Kodai hills, Tamil Nadu.

AIM: The study was conducted at Southern Regional Research Center, ICAR-Central Sheep and Wool Research Institute (CSWRI), Mannavanur, Kodaikanal, Tamil Nadu to estimate genetic trends for birth weight (BWT), weaning weight (3WT), 6 months weight (6WT), and greasy fleece weight (GFY) in a Bharat Merino (BM) flock, where selection was practiced for 6WT and GFY. MATERIALS AND METHODS: The data for this study represents a total of 1652 BM lambs; progeny of 144 sires spread over 15 years starting from 2000 to 2014, obtained from the BM flock of ICAR-SRRC (CSWRI), Mannavanur, Kodaikanal, Tamil Nadu, India. The genetic trends were calculated by regression of average predicted breeding values using software WOMBAT for the traits BWT, 3WT, 6WT and GFY versus the animal's birth year. RESULTS: The least square means were 3.28±0.02 kg, 19.08±0.23 kg, 25.00±0.35 kg and 2.13±0.07 kg for BWT, 3WT, 6WT and GFY, respectively. Genetic trends were positive and highly significant (p<0.01) for BWT, while the values for 3WT, 6WT and GFY though positive, were not significant. The estimates of genetic trends in BWT, 3WT, 6WT and GFY were 5 g, 0.8 g, 7 g and 0.3 g/year gain and the fit of the regression shows 55%, 22%, 42% and 12% coefficient of determination with the regressed value, respectively. In this study, estimated mean predicted breeding value (kg) in BWT and 3WT, 6WT and GFY were 0.067, 0.008, 0.036 and -0.003, respectively. CONCLUSION: Estimates of genetic trends indicated that there was a positive genetic improvement in all studied traits and selection would be effective for the improvement of body weight traits and GFY of BM sheep.

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.

Comparative sequence analysis of double stranded RNA binding protein encoding gene of parapoxviruses from Indian camels.

The dsRNA binding protein (RBP) encoding gene of parapoxviruses (PPVs) from the Dromedary camels, inhabitating different geographical region of Rajasthan, India were amplified by polymerase chain reaction using the primers of pseudocowpoxvirus (PCPV) from Finnish reindeer and cloned into pGEM-T for sequence analysis. Analysis of RBP encoding gene revealed that PPV DNA from Bikaner shared 98.3% and 76.6% sequence identity at the amino acid level, with Pali and Udaipur PPV DNA, respectively. Reference strains of Bovine papular stomatitis virus (BPSV) and PCPV (reindeer PCPV and human PCPV) shared 52.8% and 86.9% amino acid identity with RBP gene of camel PPVs from Bikaner, respectively. But different strains of orf virus (ORFV) from different geographical areas of the world shared 69.5-71.7% amino acid identity with RBP gene of camel PPVs from Bikaner. These findings indicate that the camel PPVs described are closely related to bovine PPV (PCPV) in comparison to caprine and ovine PPV (ORFV).