<div style="text-align: justify;">Most of the common viral diseases can be diagnosed by serological assays and these assays mostly depend on the purity and quality of antibody used. &nbsp;Such specific antibodies can be generated by hybridoma technology. Alternatively, the virus-specific antibodies can be purified from polyclonal serum by immunoaffinity purification technique. Based on this immune affinity purification technique we have purified group-specific antibody against Bluetongue virus (BTV) using recombinant protein VP7 of BTV bound to polystyrene wells. Elution buffer consisting of 100 mM Glycine-HCl, pH 3.0 was found optimum for dissociation of the antibody from recombinant antigen and also to maintain the integrity of antigen. The reactivity of eluted antibody was tested in enzyme-linked immunosorbent assay (ELISA). The purified antibody will be useful in other serological assays like ELISA, fluorescent assay, and agar gel immunodiffusion (AGID) for Bluetongue disease diagnosis.</div>

Immunoaffinity purification of bluetongue virus group specific antibody using recombinant protein adsorbed to polystyrene wells

<div>Background: Effective animal vaccines against Rabies that provide a considerable duration of immunity and mass parenteral vaccination programmes remain the mainstay of canine rabies control. Animal birth control (ABC) programmes coupled with pre-exposure anti-rabies vaccination with booster dose at regular interval have been advocated as a method to control urban street male and female dog populations and ultimately human rabies in Asia. The present study is aimed to determine the persistence of antirabies neutralizing antibodies to pre-exposure vaccination in dogs and the effect of booster vaccination on pre-exposure antirabies vaccinal neutralizing antibody levels. 
 
Methods: The study was conducted in 30 non-descript dogs which were divided into two groups containing 15 animals each. Dogs in group I were given pre exposure prophylaxis with booster dose after 21 days and animals in group II were given primary vaccination without any booster dose. Serum samples were collected at 0th, 7th, 14th, 21st, 28th, 60th, 90th and 120th days post vaccination. Seromonitoring of antirabies vaccinal antibodies was done in both groups by RFFIT. 
 
Result: 80 per cent of the animals in group I were able to maintain the mean antibody titre above protective level upto 4 months successfully, whereas, 40 per cent of the animals in group II were able to maintain mean antibody titer above protective level only upto 2 months. With the inactivated vaccine, booster dose given three weeks after the primary vaccination has ensured levels of neutralizing antibodies &gt; 0.5 IU/ml for a period of at least four months.</div>

Sequential Monitoring of Pre-exposure Antirabies Vaccinal Neutralizing Antibodies in Dogs

<div style="text-align: justify;">Occurrence of Newcastle disease (ND) and characterization of the Avian Paramyxovirus-1 (APMV-1) amongst backyard fowls (Gallus gallus domesticus) and pigeon (Columba livia domestica) was studied. ND was suspected in two fowl flocks (100% morbidity, 86% mortality) and six pigeon flocks (21.68% morbidity, 14.16% mortality) with respiratory and/or enteric clinical signs in fowls and predominantly neurological signs in pigeons was observed. Cloacal swabs, tissue lysates or infected allantoic fluid could haemagglutinate (HA) chicken erythrocytes and pigeon convalescent serum inhibited haemagglutination (antibody titres &gt;1/16). A 534 bp product was amplified from clinical samples and infected allantoic fluids by Reverse Transcriptase PCR (RT-PCR) targeting a partial Fusion protein gene including its cleavage site. The fowl and pigeon isolate was found to be velogenic and lentogenic respectively by virulence characterization. Fowl isolate grown in chicken embryonated eggs (CEE) showed 10-8.68 Embryo Infectious Dose (EID50) with a Mean Death Time (MDT) of 51.43 h and the Intra-Cerebral Pathogenicity Index (ICPI) in day-old chicks was 1.51. Pigeon isolate had corresponding values of 10-7.20 EID50, MDT of 92.00 h and ICPI of 0.43. Deduced amino acid sequence at Fusion protein cleavage site showed 112R-R-R-K-R*F117 velogenic motif for the fowl isolate and 112G-R-Q-G-R*L117 lentogenic motif for the pigeon isolate. Both isolates were found to be 84.1% related phylogenetically, while fowl isolate clustered with genotype XIII, the pigeon isolate clustered with genotype II and vaccine strains of APMV-1. This is the first instance of APMV-1 characterization in the region with the possibility of a vaccine spill over in pigeons.</div>

<div style="text-align: justify;">Molecular and Virulence Characterization of Newcastle Disease Virus in Fowls and Pigeons From Jammu, India</div>

<div>Background: The present study was undertaken to diagnose and characterize canine distemper virus (CDV) isolated from dogs of southern Gujarat, India. CDV is lethal disease of canines and felines. Total of 40 different samples were collected from 18 suspected stray dogs having different clinical signs which were processed for diagnosis and characterization of CDV. 
 
Methods: All samples were processed by employing different methods like, Immunochromatography based lateral flow test (LFA), IgG based indirect enzyme linked immunosorbent assay (i-ELISA), one step RT-PCR, nested one step RT-PCR and virus isolation in MDCK cell line. Restriction endonuclease (RE) analysis was used to characterize CDV Nucleocapsid (N) gene.&nbsp; 
 
Conclusion: Only 04 samples (02 nasal and 02 ocular swabs) of 02 dogs found positive for LFA, while 14 serum samples out of 17 samples of 18 dogs found positive for IgG antibody. As all dogs were unvaccinated, serum samples found positive in IgG based ELISA considered for confirmative positive for CDV infection. Whereas 13 samples of 10 dogs found positive for one step RT-PCR and nested one step RT-PCR. In RE digestion, characteristic two bands were found. All representative CDV positive samples of 10 dogs showed characteristic cytopathic effect in MDCK cell line. On age group wise percent positivity was found 71.42 % (05/07) in 0 - &le;6 months, while 77.77% (07/09) in 6- &le;12 months of age group, whereas, both samples were found positive in 12 months and above group. Overall 77.77% (14/18) dogs found positive for CDV infection. To the best of our knowledge, this is the first report on study of CDV infection in dogs from Gujarat state, India.</div>

Serological and Nucleocapsid Gene Based Molecular Characterization of Canine Distemper Virus (CDV) Isolated from Dogs of Southern Gujarat, India

<div>Background: Kachchhi-Sindhi horses are famous for their &lsquo;Rewal chal&rsquo; (a unique style of running). These horses dominantly exist in the western-northern border of India adjoining Pakistan. The breeding tract of these horses is Surat, Navsari, Kachchh district of Gujarat and Jaisalmer-Barmer districts of Rajasthan in India. Most familiar colors in the Kachchhi-Sindhi horses are bay and chestnut. Roman nose, ears curved at tips but not touching each other, short back, short pastern length, broader hoof for better grip and docile temperament are major characteristics of these horses. 
 
Methods: The present study is an effort to phenotypically characterize Kachchhi-Sindhi horse breed based on their different biometric indices for selection and conservation of stallions and mares for breeding purposes by private breeders. Fourteen biometric indices were recorded for phenotypic characterization of each breed viz., height at wither (HW), body length (BL), heart girth (HG), neck length (NL), face length (FL), face width (FW), pole, ear length (EL), ear width (EW), foreleg length (FLL), canon, pastern, hoof length (HoL) and hoof width (HoW). On average, these horses stand 148 cm height, have a body length of ~140 cm, a heart girth of ~165 cm, an ear length of ~15 cm and a face length of ~61 cm. 
 
Result: Horse keepers sustain horses in intensive as well as an extensive system of rearing. It has also been ascertained from various sources that horse number is declining rapidly, however, breed population statistics are not available. There is, therefore, an urgent need to conserve this breed. Proper managemental practices and conservation efforts will pave the way for the multiplication of this valuable equine genetic resource of India.</div>

<div>Phenotypic Characterization of Kachchhi-Sindhi Horses of India</div>

<div>Background: The objective of this study was to find out the association of lactation numbers of crossbred cow with their serum antisperm antibody titre, if any.&nbsp; 
 
Methods: Total 163 numbers of normal breeding crossbred jersey crossbred cows belong to various lactational groups maintained in the Instructional Livestock Farm, College of Veterinary Science, Assam Agricultural University, Khanapara and various private cattle farms in Kamrup district of Assam, India were selected for the work. ELISA was carried out for detection of antisperm antibody (asp-abs) in the test sera. Data were analysed using Chi-square test. 
 
Result: In the present study, out of 163 females 94 (57.67%) animal were found to be positive reactors. The percentage of positive reactors could be detected in 1st, 2nd, 3rd, 4th, 5th and above (5th, 6th, 7th and 9th) lactational group of cows were 16 (72.73%),15 (50.00%), 23 (63.89%), 14 (43.75%), 26 (60.47%) respectively. The highest titre up to the extent of 1:320 was recorded in the sera of cows with 1st, 3rd and 4th lactations. A negative correlation (r = -0.42NS) between the numbers of lactation and 
positive reactors for antisperm antibody (asp-abs) is found in crossbred cows.</div>

Association of Lactation Numbers of Crossbred Cows with Their Serum Antisperm Antibody Titre

Background: For the development of an immunological sperm sorting technology detection of a target protein in either X or Y chromosome-bearing sperm is mandatory. Detailed study of such proteins and finding antibodies against those sex-specific and differentially expressed proteins is the most essential step to looking forward to the immunological sexing of sperm. 
 
Methods: The specific and differentially expressed plasma membrane proteins of bovine sex-sorted semen were selected after reviewing previous research publications related to the proteomic profile of bovine-sexed semen. The in-silico characteristics such as physicochemical properties, primary, secondary, tertiary structures, membrane topology, sub-cellular localisation, domain analysis, protein interactions and epitope profiling, both linear and discontinuous peptides were done for selected plasma membrane proteins.&nbsp; 
 
Result: The results showed the localisation of proteins, their structure, polarity, stability, domains, function and probable epitopes of various selected X and Y bovine sperm plasma membrane-specific and differentially expressed proteins. For X-specific protein CLRN3, 6 and 4 linear and discontinuous epitopes and for Y chromosome-specific protein, SCAMP1, 6 and 8 linear and discontinuous epitopes were predicted, respectively. The results of in-silico characterization and epitope prediction of various selected X and Y bovine sperm plasma membrane-specific and differentially expressed proteins would be helpful in the selection of some efficient antigenic proteins to be used for further X and Y sperm-specific antibody production or synthesis of antibodies artificially.

In-silico Characterization and Epitope Profiling of Differentially Expressed Plasma Membrane Proteins of &lsquo;X&rsquo; and &lsquo;Y&rsquo; Chromosome Bearing Bovine Sperm

<div style="text-align:justify">Background: Chicken represents a valuable genetic resource and protein source, it remains a potential threat to human health as they serve as a reservoir for diseases and food borne pathogens. Diversification of the poultry production is one of the viable options to enhance the food production with less susceptibility to the threats from the emerging diseases and changing climatic conditions. Ducks are promising species suitable for the diversification and will be complimentary with an adaption potential from small to commercial farming. Majority of ducks in the country are desi type with a meagre proportion of improved duck strains.&nbsp; genetic characterization by developing duck specific microsatellite markers and designing suitable improvement program are required to be undertaken at the earliest. The microsatellite markers are extensively used for assessing genetic structure, diversity, and relationships. Information about genetic diversity of indigenous duck breeds is important to design effective improvement and conservation strategies. Therefore, the current studies aim at genetic characterization and evaluation of growth, production and reproduction traits of the crosses of local duck population besides undertaking a suitable duck improvement program. 
 
Methods:&nbsp;Spectrophotometric evaluation of DNA extraction,&nbsp; Electrophoretic evaluation of DNA extraction , microsatellite primers- Presently, a panel of 10 informative duck microsatellite markers were identified from database as reported by <a href="#alyethodi_2010">Alyethodi and Kumar (2010)</a> and <a href="#huang_2006">Huang et al., (2006)</a> and used for the studies, Polymerase chain reaction (PCR) , Documentation of PCR products by Agarose Gel Electrophoresis, Metaphor agarose gel electrophoresis(MAGE)of microsatellites alleles, Determination of molecular size of microsatellite alleles and genotypes, Statistical analysis of population genetics data, Data on genotype of all experimental birds at ten microsatellites were compiled and analyzed using POPGENE&reg; 3.1 software (<a href="#yeh_1999">Yeh et al., 1999</a>) for their population genetics parameters. 
 
Results: The study was conducted at College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, Assam, India.&nbsp; In this study, the genomic DNA was isolated and characterized microsatellite loci in crossbred&nbsp; duck (Pati x White Pekin). Ten&nbsp; microsatellites were used to detect polymorphisms in 50 cross bred ducks. A total of 28 nos of alleles were observed and all loci were polymorphic. The number of alleles ranged from 2 to 5 with an average of 2.6 &plusmn;0.08 per microsatellite locus. The observed and expected heterozygosity of these polymorphic makers ranged from 0.00 to 0.42 &nbsp;with an average number of 0.047 &nbsp;and 0.41 to 0.82 with an average number of 0.57, respectively. &nbsp;The Polymorphic markers consist of observed heterozygosities of loci were less than 0.50. The polymorphism information content (PIC) of 10 loci ranged from 0.32 to 0.78 with an average of 0.477. Microsatellite markers will be useful tool for constructing the genetic linkage map of the duck as well as comparative mapping with the chicken.&nbsp; &nbsp;</div>

Characterization of Microsatellite Markers in Crossbred Ducks

<div>Background: Porcine brucellosis is a contagious and emerging zoonosis but neglected in most of the endemic countries including India. The disease in pigs is rarely reported due to non-availability of diagnostics or major focus is on bovine brucellosis. Hence, the necessity was felt to diagnose porcine brucellosis by Monoclonal antibody based ELISA, RBPT and STATS for the detection of anti-Brucella antibodies and to record spatial seroprevalence of porcine brucellosis in the country. 
 
Methods: The Rose Bengal Plate Test (RBPT), Standard tube agglutination test (STAT) and Bru Alert monoclonal antibody based c-ELISA was employed to screen 148 sera samples for the presence of Brucella antibody.&nbsp; 
 
Result: A total 148 samples were collected and tested,&nbsp; the overall prevalence of Brucella infection in pig was 43.2% per cent (64/148) by C-ELISA followed by RBPT 40% (60/148) and STAT 25% (40/148) in the study area comparatively high seropositivity was found in female animals. The agreement between the two test was excellent (Kappa = 0.167) and also, chisquare test indicated an evidence of strong diagnostic weapon for detection of swine brucellosis.</div>

Monoclonal Antibody based Blocking ELISA for Diagnosis of Brucellosis

Majority of the Kargil sheep populations are indigenous. Very scanty information is available for morphological characterization of Purky sheep population. Therefore, the present study was undertaken with the objectives to study the morpho-metric characteristics and body measurements for phenotypic characterization of the local sheep. A stratified random sample survey was conducted on various parameters (horn size, tail size, chest girth, punch girth, body length, height at wither and body weight) in seven different age groups. Data were collected randomly from 510 sheep of either sex. The local sheep population of Kargil which was selected for the present investigation is named as &lsquo;Purky&rsquo; by the name of the community that reared this breed. In Purky sheep white body colour was predominant in both the sexes. The sheep breed was of low to medium body weight. The chest girth, punch girth, height at withers of Purky sheep showed it to be medium sized. Sex had significant effect (P&lt;0.05) on height at wither only. For males the least squares means were higher for all the traits of all age groups baring few exceptions. The phenotypic correlations of age groups with different morpho-metric traits were very high. The phenotypic correlations of &nbsp;body weight with other body measurements like tail size, chest girth, paunch girth, body length and height at wither were very high. Further studies should be carried out to fully exploit their genetic potential and to register the breed and in-situ conservation of this population. 
&nbsp;

Immunoaffinity purification of bluetongue virus group specific antibody using recombinant protein adsorbed to polystyrene wells

Sequential Monitoring of Pre-exposure Antirabies Vaccinal Neutralizing Antibodies in Dogs

Molecular and Virulence Characterization of Newcastle Disease Virus in Fowls and Pigeons From Jammu, India

Serological and Nucleocapsid Gene Based Molecular Characterization of Canine Distemper Virus (CDV) Isolated from Dogs of Southern Gujarat, India

Phenotypic Characterization of Kachchhi-Sindhi Horses of India

Association of Lactation Numbers of Crossbred Cows with Their Serum Antisperm Antibody Titre

In-silico Characterization and Epitope Profiling of Differentially Expressed Plasma Membrane Proteins of ‘X’ and ‘Y’ Chromosome Bearing Bovine Sperm

Characterization of Microsatellite Markers in Crossbred Ducks

Monoclonal Antibody based Blocking ELISA for Diagnosis of Brucellosis

Phenotypic characterization of Purky sheep population of Kargil district