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Articles by K. Dhama
Total Records ( 41 ) for K. Dhama
  M. Habtamu Taddele , R. Rathore , K. Dhama and R.K. Agarwal
  The present study was conducted to evaluate the discriminating power of RAPD-PCR and PCR-RFLP to characterize Salmonella gallinarum (S. gallinarum) isolates. Fifteen S. gallinarum isolates of poultry origin recovered from various sources in different regions of India including three reference strains were characterized using RAPD-PCR and PCR-RFLP. RAPD-PCR, using 5 primers (NSC I, NSC II, NSC III, URP-6 and primer 1290), grouped S. gallinarum isolates into 4-6 RAPD types. The discrimination power (D) of NSC II, URP-6 and primer 1290 was 0.848, 0.81 and 0.695, respectively and 0.62 for NSC I and NSC III. The phylogenetic dendrogram classified the isolates based on their geographic location. However, most S. gallinarum isolates were clustered closely together. In comparative analysis NSC II primer was placed in a separate cluster. The PCR amplification of the 16s rRNA and fimH genes produced approximately 572 bp and 1008 bp product, respectively, for all S. gallinarum isolates. Restriction Endonuclease (RE) enzymes viz., EcoRI, SmaI, AluI, BgiII, MspI and HaeIII and SalI, AluI, MspI and HaeIII were used to digest PCR products of 16s rRNA and fimH genes, respectively. These restriction enzymes generated 2-4 bands of varying molecular size ranging from 40 to 410 bp and 50 to 620 bp for 16s rRNA and fimH genes, respectively. AluI targeted against the fimH gene was the only enzyme which classified S. gallinarum isolates into three groups. In conclusion, the RAPD-PCR profile proved to be used as an epidemiological typing tool since it classified the S. gallinarum isolates according to their geographic origin.
  M.H. Taddele , R. Rathore and K. Dhama
  The study was conducted from November 2008 to May 2009 to assess the antimicrobial sensitivity pattern of 22 Salmonella isolates viz., 15 Salmonella Gallinarum strains recovered from various sources of different regions of India including 3 reference strains and 5 other Salmonella enterica serovars (7 isolates) by testing with 16 different antimicrobials. All the isolates including the standard reference Salmonella strains were subjected to antimicrobial sensitivity test using 16 different antimicrobial agents by disc diffusion method. All the Salmonella Gallinarum isolates showed resistance to erythromycin and 86.7% of them were resistant to nalidixic acid. More than 53% of the Gallinarum isolates were either 100% resistant or less sensitive to the commonly used antimicrobials, kanamycin and tetracycline whereas about 93.3% of them were sensitive to gentamicin and amoxicillin/clavulanic acid. Antimicrobial sensitivity pattern for ciprofloxacin, ofloxacin, colistin and sulfa-trimethoprim was around 88.8 and 82% of the isolates were sensitive to enrofloxacin and chloramphenicol. Among other serotypes included in the study S. typhimurium showed maximum resistance against 6 antimicrobials followed by S. kastrup which was resistant to 5 antimicrobials. S. typhimurium was 100% sensitive only to ciprofloxacin. In overall, out of a total of 22 isolates tested for different antimicrobials 4/22 (18.2%) were resistant to at least one antimicrobial and the remaining 81.8% were resistant to at least two or more antimicrobials, supporting the fact for the emergence and widespread presence of multidrug resistant Salmonella species and the importance of the implementation of suitable measures to avoid indiscriminate use of antibiotics in food animals.
  A. Hansa , R.B. Rai , M. Yaqoob Wani and K. Dhama
  Bovine coronavirus (BCoV) is widespread in cattle population, resulting in heavy economic losses to both dairy and beef industry throughout the world. The syndromes associated with BCoV include winter dysentery in adult dairy cattle and respiratory and intestinal tract infections in young calves. The virus has specific tropism for intestinal and pulmonary epithelial cells. Reports regarding prevalence and molecular detection of BCoV from India are scarce. In this study, 101 fecal samples were collected from clinical cases of diarrheic calves from North Indian region covering three dairy farms of Uttar Pradesh and dead calves of post mortem facility of Indian Veterinary Research Institute. Fecal samples from all the cases were screened for the presence of BCoV by commercially available ELISA kit. Furthermore, all samples were subjected to RT-PCR for detection and confirmation BCoV. RT-PCR was carried out using two different sets of primers to amplify the conserved nucleocapsid (N) gene of the virus targeting a 407 and 730 bp fragments. An incidence rate of more than 14% (15/101) was observed with ELISA and about 20% (20/101) by RT-PCR. The present report is first in its nature regarding the detection of BCoV at molecular level in India. In conclusion, RT-PCR was found more sensitive than commercial ELISA kit for detecting BCoV in fecal samples. Further extensive epidemiological studies are suggested for the virus in the country to know the magnitude of BCoV infection in dairy calves along with isolation of viral strains and to investigate their antigenic and genetic properties.
  K. Dhama , R.V.S. Pawaiya , S. Chakraborty , R. Tiwari and A.K. Verma
  Toroviruses are responsible for causing gastroenteritis in animals and humans. These are enveloped viruses with non-segmented and positive-sense (single stranded) RNA genome of 20 to 25 kilobases, pleomorphic and are associated with diarrhea in cattle, sheep, goat, pig and other animals and also in human beings. Morphological appearance of viruses is spherical/oval, elongated or kidney shaped. These show Torovirus-like (tubular and torus nucleocapsid in the cytoplasm of infected cells) appearance under the electron microscope and are approximately 100-140 nm in diameter, surrounded by club-shaped projections of 15-20 nm in length. Clinical signs of the disease are pyrexia, diarrhoea, dehydration, lethargy and depression in calves as well adults. In calves, the virus may lead to anorexia, mucoid faeces and neurological signs like generalised weakness, paralysis, inability to stand along with trembling and sudden death. In faecal samples, these can be identified by electron microscopy. Immunological tests Include Immuno-electron Microscopy (IEM), Haemagglutination Inhibition (HI), Enzyme Linked Immunosorbent Assay (ELISA) and southern blot. The molecular assays are reverse transcription-polymerase chain reaction (RT-PCR), nested-RT-PCR and SYBR Green real-time RT-PCR. Combined use of ELISA and RT-PCR are considered is a practical approach for epidemiological studies of bovine torovirus. At present, no vaccine is available for torovirus. The only control measures available are good hygiene and sanitary conditions along with isolation of infected animals. The present review highlights the salient features of the torovirus, their epidemiology, clinical signs, diagnosis, treatment and suitable prevention and control measures to be adopted.
  K. Dhama , S. Chakraborty , R.V.S. Pawaiya , R. Tiwari and S. Kapoor
  Foamy viruses (FVs) are complex retroviruses under the genus Spumavirus of family Retroviridae. They cause induction of multinucleated giant cell formation which presents numerous vacuoles, giving the monolayer culture a foamy appearance. FVs can infect animals as well as humans. In case of the Human foamy virus (HFV), a defective variant (named ΔHFV or HFVΔTas) negatively interferes with replication of parental counterpart. Some species, such as rhesus macaques, African green monkeys, chimpanzees and cats harbor closely related yet serologically distinct FV subtypes. Unanticipated FV pathogenicity may warrant appropriate attention to biosafety practices to prevent occupational infections and the importance of additional studies to better define clinical outcome of these zoonotic infections. During cross-species infection and subsequent passages a rapid and fatal disease can occur, with changes from nonpathogenic to pathogenic potentials. In persons occupationally exposed to non-human primates, Simian foamy virus (SFV) infection occurs persistently showing that simian retroviruses cross into humans more frequently. Simian Immunodeficiency Viruses (SIV), mostly are nonpathogenic in their natural hosts but during cross-species infection a rapid and fatal disease can occur. Enzyme Immuno Assay (EIA), Western blot analysis and Polymerase Chain Reaction (PCR) amplification are the important diagnostic tests for FVs. FVs are also being exploited as potential vectors that can be used for gene therapy which is gaining much attention of the researchers worldwide. Strengthening sero-epidemiological as well as molecular investigations and public health surveillance programme along with extra precautions while transferring xenograft are some of the approaches to prevent these viral infections.
  K. Dhama , R.V.S. Pawaiya , K. Karthik , S. Chakraborty , R. Tiwari and A.K. Verma
  Equine Encephalosis (EE) is an arthropod borne febrile non contagious disease of equines. The causative virus, Equine encephalosis virus (EEV), has several serotypes (EEV1-EEV7) and the virus has been reported from southern Africa including Kenya, Botswana and South Africa. EEV was first isolated in 1967 from horses in the Republic of South Africa. Like the African horse sickness virus (AHSV) EEV is also endemic in southern Africa. In most of the country, EE virus in comparison to AHSV has a higher transmission rate. Two species in the Culicoides imicola species complex, C. imicola (senso stricto) and C. bolitinos are known to transmit EEV. Zebra and elephants can act as maintenance host of the virus, making the elimination of the virus difficult. Outbreaks of EEV infection is reported to be associated with equine foetus abortion during the first 5-6 months of gestation. 32P-labelled genomic probes of EEV are used for detection of viral Ribonucleic Acid (RNA). Sero-epidemiological tools for the detection of antibodies against EEV include Serum Neutralization Test (SNT) and Enzyme Linked Immunosorbent Assay (ELISA). A novel real time Polymerase Chain Reaction (PCR) assay has also been developed for the detection of EEV by targeting the gene Viral Protein (VP)-7. There is no specific treatment or vaccine available for this virus. Supportive treatment can only be provided. Management of horses in the stable is the key to control the spread of EEV in equines along with follow up of good biosecurity measures. The present review deals with all these aspects of the infection caused by this virus to enrich knowledge of researchers and equine/stud farm owners and the industry.
  P.K. Nanda , P. Swain , S.K. Nayak , T. Behera and K. Dhama
  Enzymatic method using trypsin to establish primary culture from various cells from heart, liver, kidney, brain, testis, ovary, fin and embryo of Indian Major Carp, Cirrhinus mrigala was investigated and compared with that of explant culture method. The dissociation time to obtain cells from individual tissues and the viability of cells for primary culture varied at 0.125 and 0.25% trypsin concentration. Single cell culture of trypsinised cells from testis, ovary and embryo showed good attachment irrespective of trypsin concentration. Cells from liver and heart were found to be sensitive to trypsin only at higher concentration (0.25%); whereas, no attachment/or proliferation was observed from cells of kidney, brain and fin due to sensitivity to trypsin at both the concentrations. In contrast to this method to obtain primary culture, explant culture from most of the tissues showed better attachment, resulting in subsequent growth and proliferation of cells forming monolayer. Overall, the explant culture of most of the tissues of C. mrigala was found to be suitable and survived more passages as compared to single cell culture obtained through trypsinisation. For obtaining primary culture from single cells, further investigations are desired to identify tissue specific enzymes, standardization of dose, duration and temperature of enzymatic treatment.
  S. Aravind , N.M. Kamble , S.S. Gaikwad , S.A. Khulape , Sohini Dey , K. Dhama and C. Madhan Mohan
  The aim of the study was to identify the codon usage bias between the newly identified Duck Enteritis Virus (DEV) gD gene (GenBank accession No. KC915041) and the gD like gene of 23 other reference herpesviruses. The codon usage bias analysis of gD gene of DEV may improve our understanding of the evolution and pathogenesis of DEV and provide a basis for understanding the relevant mechanism for biased usage of synonymous codons; and for selecting appropriate heterologous expression systems to improve the expression of target genes. The results showed that codon of gD gene of DEV was having strong bias towards the synonymous codons with A and T at the third codon position. A high level of diversity in codon usage bias existed; and the effective number of codons used in a gene plot revealed that the genetic heterogeneity in gD gene of herpesviruses was constrained by the G+C content. The phylogenetic analysis suggested that DEV was evolutionarily closer to Alphaherpesvirinae, there was no significant deviation in codon usage in different virus strains. There were 17 codons showing distinct usage differences between DEV and Escherichia coli, 22 between DEV and Homo sapiens but only 15 codons between DEV and yeast. Therefore, the yeast expression system may be more suitable for the expression of DEV genes. The results are encouraging regarding bioinformatics data of an economically important poultry pathogen and could be highly useful for development of new generation vaccines, diagnostics and studying the evolution of the duck enteritis virus.
  K. Dhama , R.P. Singh , K. Karthik , S. Chakraborty , R. Tiwari , M.Y. Wani and J. Mohan
  Spallanzani’s thought of Artificial Insemination (AI) has revolutionized the animal husbandry field, both in developing and developed countries, by improving the genetic potential of livestock and poultry; minimizing the managemental costs and holding the service of genetically superior males even after their death. AI in domesticated birds especially in turkey shows promising results unlike other domestic and wild animals. The advantages of AI are many which support the wide adaptation of this technique in the poultry industry to augment its growth. Making AI as an integral part of captive breeding programme for non-domesticated birds would facilitate the process of saving various endangered species of wild birds. However, there are various problems involved in case of birds which need to be addressed before implementing AI. Apart from these, AI also poses a risk of possible transmission of various infectious pathogens/diseases of poultry through semen or its contamination or during the process of insemination. Hence, careful and regular screening and monitoring of poultry will help to check the spread of such diseases. Novel methods are adopted to prevent the colonization of contaminant microbes in stored semen thereby minimizing the pathogen transfer. The recent advances in biotechnology and molecular biology need to be explored fully for early and rapid diagnosis of poultry diseases. This would help in formulating appropriate disease prevention and control strategies and thus safeguard poultry health and production. This review describes the salient facts about AI practices in poultry and possible transmission of infectious pathogens during insemination along with suitable prevention and control strategies to be adapted.
  K. Dhama , R.V.S. Pawaiya , S. Chakraborty , R. Tiwari and A.K. Verma
  Powassan encephalitis is a rare but severe disease caused by infection with Powassan virus (POWV). It is a tick-borne Flavivirus (family Flaviviridae) having single stranded Ribonucleic Acid (RNA) which is positive sense in nature. The virus has highest case-fatality rates and is associated with a very high incidence of severe neurologic sequelae. Humans contract POWV infection accidentally when they are exposed to areas where the virus, arthropod vector (an Ixodid tick) and the vertebrate natural hosts coexist. Reported incubation periods for Powassan virus range from 8 to 34 days. The disease is associated with a reactive inflammatory cellular infiltrate (chronic) of lymphocytes and macrophages that include the abundance of perivascular inflammatory cells and multiple foci of parenchymal cells in grey matter. Basically two diagnostic approaches are considered. First one is the direct detection of the virus or viral RNA in the initial (viremic) phase of infection by virus isolation in mammalian cell culture or by reverse transcriptase polymerase chain reaction (RT-PCR). Second is the indirect detection of specific immunoglobulins (IgM and IgG antibodies) with serological methods such as Enzyme Linked Immunosorbent Assay (ELISA); Immunofluorescence Assay (IFA) or Neutralization Tests (NTs). Phylogenetic analysis is important for genogrouping of the virus. Oligomers targeting specific locations in the RNA genome of the flavivirus have been used at present for successful suppression of viral gene expression. Strict hygienic and biosafety measures including tick control is pre-requisite for prevention of disease. The present review will give an insight to the details of disease caused by this arbovirus that may often prove fatal, its epidemiology, diagnosis, prevention and control measures to be adopted.
  K. Dhama , R.V.S. Pawaiya , S. Chakraborty , R. Tiwari , M. Saminathan and A.K. Verma
  Coronaviruses are positive-sense single-stranded ribonucleic acid (RNA) viruses causing a broad spectrum of diseases in domestic and wild animals including poultry and rodents. Based on antigenic and genetic similarities coronaviruses have been subdivided into 3 major antigenic groups. They infect and produce disease in multiple species of animals, human beings (group 1 and 2) and birds (group 3). Equine coronavirus (ECV) causes enteritis in foals. Complete genome of first ECV isolate NC99 strain has been recently sequenced. Cytolytic nature of the virus is responsible for occurrence of lesions in the small intestine, thereby causing diarrhea. Demonstration of Coronavirus antigens in clinical samples is test of choice for diagnosis. By electron microscopy (negative staining) Coronavirus like particles can be identified in fecal samples. Coronavirus antigen in fecal samples can be detected by antigen capture enzyme linked immuno-sorbent assay (ELISA). Molecular detection tool like reverse-transcriptase polymerase chain reaction (RT-PCR) has made the diagnosis more accurate. Virus characterization along with genogrouping has become easier these days with the advent of proteomics and phylogenetic studies. Currently, no vaccine is available for ECV. Biosecurity measures if adopted strictly prevent the disease. The present review highlights the salient features of the Coronavirus in general with special reference to ECV and the disease it causes in equines, its epidemiology, diagnosis and appropriate prevention and control measures to be adopted. The review would be helpful for understanding the virus/disease in a better way and alleviating economic losses to the equine/stud farm owners.
  K. Karthik , R. Rathore , P. Thomas , A. Elamurugan , T.R. Arun and K. Dhama
  Brucella abortus, one of the major pathogen causing abortions in cattle worldwide and a zoonotic agent, need to be detected earlier in order to prevent its spread among animals. The present study aimed at to know the prevalence of B. abortus in cattle population of three states (Uttar Pradesh, Uttarakhand and Tamil Nadu) of India by serological (Rose Bengal Plate Test (RBPT) and Serum Tube Agglutination Test (STAT)) and molecular (polymerase chain reaction) detection in sera samples and whole blood (n = 370), respectively. Out of a total of 370 sera samples, 61 (16.49%) were positive by RBPT and 59 (15.94%) by STAT. Screening of the whole blood samples by genus specific bcsp31 gene based PCR as well as species specific IS711 gene based PCR revealed that 56 (15.13%) samples were positive for brucellosis. None of the serologically negative sample showed positivity by PCR; however few positive samples were tested negative by PCR. Sensitivity and specificity of PCR compared with RBPT was 100 and 92.4% while with STAT these were 100 and 95.16%, respectively. Results are promising that whole blood can be used for studying the molecular epidemiology of B. abortus in cattle and particularly detecting the active phase of infection and PCR can be well adopted as a valuable test for mass screening of animals for this purpose. The present study adds to the prevalence data available regarding to B. abortus infection in cattle population and highlights the usefulness and advantages of molecular tool of PCR over serological tests.
  T.R. Arun , R. Rana , P. Singh , P. Choudhuri , V.P. Singh , P. Thomas , V. Rekha , K. Nehra , J. Usharani and K. Dhama
  A gold nanoparticle based lateral flow assay was developed for rapid serodiagnosis of contagious agalactia, an economically important mycoplasmal disease of small ruminants. Sonicated antigen of Mycoplasma agalactiae was used as the test reagent that was immobilized on nitrocellulose membrane along with the control line of goat IgG. The detection reagent, gold nanoparticle conjugated with anti-goat antibody was dried on the conjugate pad. During the assay, specific antibodies against M. agalactiae in the test serum that combined with the detection reagent were captured in the test line and detected visually by the development of a red line on nitrocellulose membrane. The gold conjugate captured in control line produced a red line regardless of the presence of specific antibodies that served as a procedural control. Serum samples collected from an experimentally infected goat were tested with the lateral flow assay and antibodies were detected from 9th day of infection and the assay was also evaluated using 100 goat sera samples. This is the first report regarding development of a gold nanoparticle based lateral flow assay for rapid diagnosis of contagious agalactia in goats. This study suggests that current lateral flow assay can be used as a user friendly diagnostic in laboratories lacking specialized equipments as well as for point of care diagnosis of contagious agalactia.
  P. Thomas , T.R. Arun , K. Karthik , P.V. Berin , M. Asok Kumar , Neetu Singh , J. Usharani , M. Palanivelu , S.K. Gupta , K. Dhama and K.N. Viswas
  Necrotic enteritis, caused by Clostridium perfringens, is an important bacterial disease of poultry. A suspected case of necrotic enteritis was presented for necropsy from an Indian Kadaknath Fowl flock showing diarrhea and progressive debility. Gross examination revealed necrotic to ulcerative lesions in intestine. The organism was isolated from the intestinal contents, tissue and liver under anerobic conditions. The cultural characteristics and Gram staining were suggestive of C. perfringens. The sequencing of 16s rRNA gene confirmed the isolate as C. perfringens and which was well differentiated from other clostridia associated with avian intestinal tract. This study demonstrates that 16s rRNA gene sequencing can provide rapid and confirmatory identification of C. perfringens. Further, Multiplex Polymerase Chain Reaction (mPCR) was performed for toxinotyping and isolate was found to be positive for α toxin (cpa) and β2 toxin (cpb2), a feature of C. perfringens type A isolates. As some recent studies have highlighted the involvement of NetB toxin in pathogenesis, therefore, PCR was carried out to find the presence of this toxin, the isolate was found to be negative for netB gene. This study emphasizes the molecular characterization and toxinotyping as a rapid tool for detection of C. perfringens from suspected necrotic enteritis cases. Very few reports regarding molecular characterization are available from India, hence it adds to the available data on this important poultry pathogen. Further investigations are required to understand the exact role of NetB toxin in pathogenesis as various studies including the current one reports NetB negative strains involved in necrotic enteritis.
  Anjaneya , S.D. Singh , K. Dhama , M.Y. Wani , V. Gowthaman and M.M. Chawak
  Infectious Coryza (IC) is one of the highly infectious and contagious respiratory tract disease of poultry caused by Avibacterium paragallinarum. It has emerged as one of the major problem of commercial poultry industry worldwide due to huge economic losses in terms of increased number of culls and significant reduction in egg production (10-40%). Scarce reports are available regarding prevalence of Av. paragallinarum from India. The present study was designed to characterize the Indian Av. paragallinarum serovars at molecular level based on the Restriction Fragment Length Polymorphism (RFLP) of 16S ribosomal gene amplified, amplified 16S ribosomal DNA restriction analysis (ARDRA), sequencing of haemagglutinin antigen (hagA) gene and phylogenetic analysis. Four culture positive isolates of Av. paragallinarum, recovered from different geographical locations of the country, were confirmed by species specific HPG-2 Polymerase Chain Reaction (PCR) and other 10 nasal sinus swabs were found positive by direct HPG-2 PCR. ARDRA technique amplified product of 16S ribosomal DNA (r-DNA) showed identical Restriction Enzyme Analysis (REA) pattern for all the isolates indicating presence of similar operons in 16S ribosomal gene. The ‘hagA’ encoding a haemagglutinin antigen of Av. paragallinarum from all the field isolates revealed homology between Australian A and C serovar strain but did not cluster according to Page’s serovar scheme of classification. The present study is first in its nature regarding the molecular characterisation of Indian field isolates of Av. paragallinarum and provides valuable information regarding their genetic nature. Further extensive molecular studies will help both in the identification of field isolates and devising appropriate prevention and control measures for this important pathogen of poultry.
  P.K. Nanda , P. Swain , S.K. Nayak , T. Behera , P. Jayasankar and K. Dhama
  The success in establishing in vitro culture from anchorage dependent tissue explants depends upon their ability to attach to culture substratum. In this study, evaluation of different coating factors on the attachment and subsequent monolayer formation from various tissue explants of Indian Major Carp, Mrigal, Cirrhinus mrigala was investigated to establish cell culture. Explants from heart, liver, gills, testis, ovary, kidney and fin were taken in respective culture flasks coated either with type I collagen, gelatin or fibronectin to know their ability on attachment and proliferation of cells resulting in attaining confluency. The percentage attachment of explants from same tissue, when cultured in different coated flasks, varied greatly. Explants (40-65%) from liver, ovary, testis and fin were found to attach well in fibronectin coated flasks, whereas maximum attachment (>65%) from heart tissue was recorded in gelatin coated flasks. This difference in percentage attachment of explants to substratum significantly affected growth, proliferation of cells and subsequent formation of confluent monolayer. Overall, fibronectin as a coating factor was found to be good for attachment of explants from most of the tissues of C. mrigala. Contrary to this, attachment percentage of explants from fin, gills, ovary and testis in type I collagen coated flasks was less which eventually failed to form monolayer. The findings indicate that coating factor(s) has a major role in establishing cell culture which not only influences attachment but also growth and proliferation of cells from explants obtained from different tissues of C. mrigala. Further investigations are suggested to find out suitable coating factors for each cell types of different fish species so as to establish cell cultures.
  Rupasi Tiwari , H. Dileep Kumar , Triveni Dutt , B.P. Singh , K. Pachaiyappan and K. Dhama
  Today about 7.08 billion people inhabit the earth. Presently to feed every one adequately the world needs 2800 million tonnes of cereals, against which global production is only 2100 million tonnes. This discrepancy in need and production has left over 868 million people undernourished worldwide and 850 million of them live in developing countries. On the other hand rapid urbanization, increasing purchasing power and shift in diet is pushing demand for richer diets and protein of animal origin. By 2050, consumption of meat and dairy products not the butter is projected to increase by 173 and 158%, respectively, as that of 2010. To meet the growing demand and to cope with 9 billion world population by 2050, agricultural production needs to increase by 60% (compared to 2005/2007 production) including of increase in animal production and animal products. Feeding this burgeoning population with the available 0.23 ha of cropland per capita is one of the biggest challenges. Apart from this, the world is facing another biggest challenge of climatic change. Simply, if the “business as usual scenario” continues it requires three earth planets to supply the resources for human population consumption and to assimilate associated wastes it generates in 2050. In the present review, an attempt has been made to throw light on the future challenges of population rise, limited natural resources and climate change to ensure food security in India and the world with major emphasis on sustainable livestock production.
  S.H. Somshekhar , B.M. Veeregowda , V.V.S. Suryanarayana , G. Leena , K. Dhama and S. Chakraborty
  Haemorrhagic Septicaemia (HS) is an acute fatal septicaemic disease of cattle and buffaloes. It is caused by Pasteurella multocida serotype B:2. The disease is of great economic importance in India mainly due to the high mortality in susceptible populations. Bacterial Outer Membrane Proteins (OMPs) play an important role in infectious process of many bacteria. In the present study, OMP of P. multocida (serogroup B) field isolates (n = 12) and a vaccine strain (P-52) were extracted by a sarkosyl method and characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. A total of 9-14 different polypeptide bands were observed with approximate molecular weight ranging from 16-123 kDa. On the basis of stain intensity, 32 kDa protein appeared to be the major protein band followed by 37, 26, 29, 89 and 72 kDa bands. Two protein bands of 123 and 46 kDa were present only in vaccine strain. Protein band of 69 kDa was present only in four of the field isolates and vaccine strain whereas OMP with 43 kDa was detected in only four of the field isolates and absent in remaining field isolates and vaccine strain. Immunoblotting identified 32, 37, 72 and 89 kDa protein bands as immunogenic OMPs. It can, therefore, be concluded that outer membrane proteins play a significant role in the pathogenesis of pasteurellosis. Several OMPs act as immunogens for which the antibodies produced against them demonstrate a strong protective action and such OMP antigens may be used as component of subunit vaccines.
  D. Neeraja , B.M. Veeregowda , M. Sobha Rani , D. Rathnamma , H.D. Narayanaswamy , M.D. Venkatesha , G. Leena , R. Apsana , S.H. Somshekhar , M. Saminathan , K. Dhama and S. Chakraborty
  Bovine tuberculosis (bTB) is an economically important zoonotic disease (can spread to human through inhalation or ingestion) caused by Mycobacterium bovis which belongs to Mycobacterium tuberculosis complex (MTC). Control and eradication of infection is difficult even in organized dairy farms. So combinations of tests like culturing and nucleic acid-based diagnostics are used for the isolation and identification of mycobacterial infections in cattle. Even though, there are many advances in diagnosis of bovine TB infection in cattle but till now, isolation identification of the etiological agent from clinical samples stands as a definitive and gold standard test. Nucleic acid based methods like Polymerase Chain Reaction (PCR) which have advantages of speed, sensitivity and specificity can be used for diagnosis of tuberculosis along with isolation. In the present study, isolation of Mycobacterium tuberculosis complex organisms was attempted from nasal swabs and milk of cattle using Lowenstein-Jensen (LJ) media without glycerol. Cattle which were positive for tuberculosis either by skin test or gamma interferon test were selected. Two of the twelve nasal swabs and none of the seven milk samples showed typical mycobacterial colonies on LJ media after 8 weeks of incubation. Ziehl-Neelsen staining of colonies showed slender, rod shaped acid fast organisms suggestive of Mycobacterium. Deoxy ribo nucleic acid (DNA) was extracted by boiling method and amplified by duplex PCR for 245 and 500 bp amplicons specific for MTC (IS6110) and M. bovis (RvD1Rv2031c), respectively. Electrophoresis revealed 245 bp product but not 500 bp which confirmed the identity and relatedness of the isolated mycobacterium to Mycobacterium tuberculosis complex.
  S.A. Bhat , J.K. Khajuria , R. Katoch , M.Y. Wani and K. Dhama
  Backyard poultry rearing is an important venture and integral part of mixed farming in most of the developing countries of world. Very scarce reports regarding the effects of parasites on free ranging birds are available from India. The present study was designed to investigate the prevalence of helminth parasites in the backyard poultry farming in the northern, humid and subtropical region of India. A total of 120 gut specimens and 600 faecal samples of backyard poultry were collected from different villages and analysed for parasitic worm loads and different egg/ova types. Furthermore, a random field trial on 40 birds reared in backyard poultry system was carried out to determine the effect of fenbendazole treatment on the parasitic load and production performance. On gut examination, the most common nematodes found were, Ascaridia galli (20%), Heterakis gallinarum (10.83%), Capillaria spp. (5%) and Cheilospirura hamulosa (1.67%) while the cestodes were Raillietina tetragona (9.16%), R. echinbothrida (5%), Hymenolepis spp. (5%), Cotugnia digonopora (3.33%) and R. cesticillus (2.5%). The faecal examination showed higher incidence of A. galli (19.16%), H. gallinarum (9.5%), Capillaria spp. (3.5%), Trichostrongylus tenuis (2.5%), Raillietina spp. (16.16%), Eimeria spp. (5.33%) and mixed infections (6.67%). Treatment with fenbendazole was found to reduce mortality (15%) as compared to untreated (30%) groups. Moreover, fenbendazole treated birds gained significantly steadily weights (r = -0.35) as compared to untreated group (r = -0.019). The study is first in its nature in providing the valuable information regarding prevalence of endoparasites based on faecal examination in backyard poultry from the Jammu region. This information will essentially be helpful for the researchers and local veterinarians to develop strategies for both treatment and control of these endoparasites affecting poultry.
  D. Neeraja , B.M. Veeregowda , M. Sobha Rani , D. Rathnamma , R. Bhaskaran , G. Leena , S.H. Somshekhar , M. Saminathan , K. Dhama and S. Chakraborty
  Bovine tuberculosis (bTB), caused by Mycobacterium bovis which belongs to Mycobacterium tuberculosis complex (MTC), is a globally distributed zoonotic disease in cattle. The present study was conducted in a dairy herd with the history of prevalence of bovine tuberculosis. Two Cell Mediated Immunity (CMI) based tests, Single Intradermal Test (SID), gamma interferon (IFN-γ) assay and a serological test (enzyme linked immunosorbant assay, ELISA) were employed for the diagnosis of tuberculosis in 45 animals. Of these, 8 (17.77%) were positive by SID test, 10 (22.22%) by interferon gamma assay and none of the animals were found positive by ELISA. Both the CMI tests performed were found better than the antibody detection. Between the CMI tests, IFN-γ assay showed better sensitivity than the SID test; combination of tests showed better detection of the bTB infection in animals. ELISA results indicated that animals were still in progressive stages of infection and the sensitivity of the tests depends on the stage of infection in the study subjects. As a whole, 26.67% of the animals tested in the farm were found to be positive and/or reactors to tuberculosis. It indicates that both the CMI tests were better than those targeting antibody detection. ELISA did not detect even a single animal as positive. These results indicate that no single test is able to detect all the infected animals and also not having 100 % sensitivity and specificity. So combination of tests always can be better employed for the diagnosis of bovine tuberculosis.
  P.L. Lalruatfela , M. Saminathan , R.S. Ingole , K. Dhama and M.V. Joshi
  Paraquat (PQ) is commonly used in agriculture as a contact herbicide to control the growth of weeds and grasses and is easily accessible to farmers in developing countries. Paraquat is highly toxic to human and is associated with high mortality varying from 35-50%. The main causes for mortality are respiratory and multi organ failure. In this experimental study, twenty four female Wistar rats of 6-7 weeks age were divided into four equal groups. Group-I rats served as control whereas groups-II, III and IV rats were given daily paraquat solution at the dose rate of 10, 15 and 25 mg kg-1 b.wt, respectively, orally by gavage for 28 days. At end of the 2nd week of the experiment, mild diarrhoea, anorexia, polydipsia and reduced locomotional activities were noticed. Hematological observations showed significant decrease in Hb, PCV, TEC and TLC values and biochemical parameters revealed significant increase in AST, ALT and creatinine levels when compared to control group. Gross pathological observations of lungs from all treatment rats showed mild congestion and emphysema. An accidental finding of hydronephrosis was recorded in group-II and atrophy of left kidney and hypertrophy of right kidney was observed in group-III. Few animals from group-IV showed rounded flabby heart. Histopathologically, granular and vacuolar changes were observed in liver and kidneys. Lungs showed prominent congestion and oedema. From the present investigation it was concluded that paraquat toxicity adversely affects general performance, hematological and biochemical parameters in rats. Histopathological changes in liver and kidneys indicated its hepatoxic and nephrotoxic effects.
  K.C. Das , R. Sharma , S.S. Paul , K. Dhama , N. Haque , K.K. Baruah and C. Rajkhowa
  Six fibre degrading fungus were isolated from rumen of mithun (Bos frontalis) at National Research Centre on Mithun, Jharnapani, Medziphema, Nagaland using specific media for rumen fungus. These isolated fungal cultures showed higher fibre degrading enzymes like CM cellulase, Xylanase and Avicelase activity. Addition of fungal culture in the mixed rumen liquor of mithun increased in vitro gas production. The identification and characterization of these isolates was done by DNA isolation, Polymerase Chain Reaction (PCR) amplification, sequencing and plotting of phylogenic tree. The result showed that two isolates were homologus to Orpinomyces species and four isolates were homologus to Neocallimastix species. These isolates can have potential to be used as microbial feed additives in ruminants.
  P.S. Bagalakote , R.S. Rathore , T.P. Ramees , H.V. Mohan , M. Sumankumar , R.K. Agarwal , A. Kumar and K. Dhama
  In modern years, Arcobacters are reflected as potential emerging food-borne zoonotic entero-pathogens. Arcobacter species displayed a wide variety of genetic diversity. The study was carried out to genotype and find molecular heterogeneity of Arcobacter spp. (Arcobacter butzleri, A. cryaerophilus and A. skirrowii), isolated from different sources from Bareilly region, Uttar Pradesh, India by using randomly amplified polymorphic DNA - polymerase chain reaction (RAPD-PCR). RAPD-PCR was performed using genomic DNA of Arcobacter isolates (n = 56; 33 A. butzleri, 20 A. cryaerophilus, 3 A. skirrowii; recovered from chicken meat, pork, sheep faeces, goat faeces, poultry intestinal contents and human diarrhoeal stool samples) as template by employing two published primers. The RAPD profiling for primer 1 (HLWL85) yielded number of bands rangeing between 2-8 (500-3100 bp). Out of 56 isolates, 54 showed bands giving a typeability of 96.4%. These 54 typable strains were grouped to 35 types and giving discriminatory power of 0.9762. Primer 2 (OPA-11) yielded RAPD-PCR profiles comprising of 2-7 bands (210-2800 bp). Out of total 56 isolates, 54 were typable with a discriminatory power of 0.9336. This is the first report from India regarding RAPD profiling of Arcobacter spp. This study reveals epidemiological relationship of Arcobacter isolate from various sources and will help to design suitable prevention and control strategies for this important pathogen having public health significance.
  P. Mohapatra , R.K. Swain , S.K. Mishra , T. Behera , P. Swain , N.C. Behura , G. Sahoo , K. Sethy , B.P. Bhol and K. Dhama
  To study the comparative effects of Nano Selenium (Nano-Se) and sodium selenite on the growth, bioavailability, antioxidative activities, hematological and biochemical parameters, cellular and humoral immunity, a trial was conducted on BV 300 layer birds during grower phase (9-20 after weeks) in six treated groups. After twenth weeks body weight of the all Nano-Se treated groups (up to a dose of 0.3 mg kg-1 of diet) was found to be significantly (p<0.05) higher than sodium selenite treated and control groups. However, further increase in dietary Nano-Se content in feed had negative effect on body weight of bird. Birds fed with both Nano-Se and sodium selenite showed higher (p<0.05) Se content in different tissues (Such as breast muscle, liver, kidney, pancreas, serum and feathers). However, Se content in liver, breast muscle, pancrease and feathers were signifcantly higher (p<0.05) in Nano-Se treated groups. In addition, significant (p<0.05) difference was observed as regard to glutathione peroxidase (GSH-Px), erythrocyte catalase (CAT) and superoxide dismutase (SOD) activities among the treated groups. Significantly better cellular and humoral immunity response were observed in Se supplemented birds. Dietary supplementation of Nano-Se improved the body weight, feed consumption ratio, antioxidant status, immunity and tissue Se deposition in grower birds.
  M. Saminathan , R.B. Rai , K. Dhama , G.J. Ranganath , V. Murugesan , K. Kannan , S. Pavulraj , A. Gopalakrishnan and C. Suresh
  Mammary tumours rank second as the most common neoplasms in dogs after skin tumours, whereas in women the most common cause of cancer-related deaths is breast cancer. N-Methyl-N-Nitrosourea (NMU) is a highly specific mammary gland carcinogen which directly act and does not require metabolic activation. In the present study, NMU at the dose rate of 50 mg kg-1 body weight was used intra-peritoneally for the induction of mammary tumour. The first palpable tumour appeared on 70th day post carcinogen injection and subsequently, most of the tumours were developed around 18-20th week. During 28 weeks of experimental period, the tumour incidence was 82.86% (29/35). The tumour frequency was found to be 4.7±0.33 tumours and the average latency period was 107±4.1 days. The average tumour volume was found to be 69±8.8 cm3. Equally, 50% of mammary tumours appeared on the right (22/44) and left (22/44) mammary gland chain. Region wise, 81.82% (36/44) of the tumours appeared on abdominal-inguinal mammary glands and 18.18% (8/44) on the cervical thoracic mammary glands. A total of 44 mammary tumours were diagnosed in which 88.64% (39/44) were malignant and 11.36% (5/44) were benign. Among the malignant tumours, 33.33% (13/39) were non-invasive and 66.67% (26/39) were invasive. The average values of mitotic index, Proliferating Cell Nuclear Antigen (PCNA), Vascular Endothelial Growth Factor (VEGF) and Platelet Endothelial Cell Adhesion Molecule (PECAM-1) in NMU induced mammary tumours were found to be 4.5±0.46/hpf, 77±2.6, 16.2±0.86 and 15±0.69, respectively. The present study for the first time demonstrated the expression of VEGF and PECAM-1/CD-31 proteins in NMU induced mammary tumours.
  Manoj Jinu , R.K. Agarwal , B. Sailo , M.A. Wani , Ashok Kumar , K. Dhama and M.K. Singh
  The aim of the study was to compare Polymerase Chain Reaction (PCR) and conventional method for detection of Salmonella from field poultry samples (n = 510, poultry blood and faeces 255 each). The prevalence rate of Salmonella in chicken was found to be 5.09% using conventional method and 5.88% by PCR assay. Serotyping of 26 Salmonella isolates revealed 57.69% Salmonella Typhimurium, 19.23% rough type, 15.38% Salmonella Enteritidis and 7.69% untypable. Among Salmonella Typhimurium isolates, 73.33% were from poultry blood and 26.66% from faeces samples. All isolates belonging to Typhimurium and Enteritidis serotypes were confirmed by PCR targeting of Salmonella Typhimurium (typh) and Salmonella Enteritidis (ent) specific genes. However, 4 isolates found to be rough type also turned out to be positive for ent gene. The PCR employed for detection of Salmonella was found 100% sensitive for poultry blood but its sensitivity was very less (77.77%) for faeces samples as compared with culture method. However, PCR was 100% specific with regard to faeces samples. The specificity from blood samples was 97.89% by PCR. The positive predictive values of PCR from blood and faecal samples were 77.27 and 100% with a concordance of 98.03 and 99.21%, respectively. The negative predictive values from blood and faecal samples were 100 and 99.19%. The study demonstrated usefulness of genus specific PCR for detection of Salmonella in poultry clinical samples. Owing to its robustness and rapidity it can be used for wide epidemiological studies. Serotype specific PCR detection of Typhimurium and Enteritidis serotypes has added advantage in identifying them even where there is loss of O antigen.
  P. Mohapatra , R.K. Swain , S.K. Mishra , T. Behera , P. Swain , S.S. Mishra , N.C. Behura , S.C. Sabat , K. Sethy , K. Dhama and P. Jayasankar
  Comparative study on the effect of nano selenium (nano-Se) and sodium selenite on the growth, bioavailability, antioxidative activities, hematological and biochemical parameters, cellular and humoral immunity was done in layer chicks upto 8th week post feeding. The results showed significant differences (p<0.05) in relative weight gain and final body weight of the nano-Se treated groups upto a dose of 0.3 mg kg-1 of diet as compared to sodium selenite and control groups. However, further increase in dietary nano-Se content in feed had negative effect on weight and Relative Gain Rate (RGR). Survival rate and Feed Conversion Ratio (FCR) were not affected by dietary treatments. Chicks fed with both nano-Se and sodium selenite showed higher (p<0.05) Se content in different tissues (breast muscle, liver, kidney, pancreas, serum and feathers). However, highest value (p<0.05) of Se content in breast muscle and liver was observed in nano-Se treated groups. Selenium concentrations in serum, liver and breast muscle increased linearly and quadratically (p<0.05) as dietary Se level increased for all Se sources but its magnitude was substantially greater (p<0.05) when nano-Se was fed. Glutathione peroxidase (GSH-Px), erythrocyte catalase (CAT) and superoxide dismutase (SOD) activities were significantly different (p<0.05) in all treated groups than control. Dietary nano-Se also increased several serum biochemical and haematological parameters. In addition, it significantly increased both cellular and humoral immunity in layer chicks after 8th weeks of post feeding. In conclusion, dietary administration of nano-Se was found superior than that of inorganic sodium selenite in various aspects in layer chicks. Further extensive study for exploring absorption mechanisms, metabolic pathways, ideal dose/form of nano-Se is suggested for optimum utilization of nano-material based application of Se feeding in poultry.
  A. Das , S.K. Mishra , R.K. Swain , G. Sahoo , N.C. Behura , K. Sethi , B. Chichilichi , S.R. Mishra , T. Behera , K. Dhama and P. Swain
  A comparative study on effect of replacement of inorganic minerals viz., Zinc (Zn), Copper (Cu) and Manganese (Mn) with their corresponding organic minerals (methionine) on growth, bioavailability and immunity in layer chicks was undertaken till 8th week post feeding. At 8th week, the body weight of birds were found to be significantly (p<0.05) higher in 100% organic Zn group, 100 and 50% organic Zn, Cu and Mn supplemented groups. The cumulative feed consumption and Feed Conversion Ratio (FCR) of all the treated groups showed no significant (p>0.05) difference. The serum glucose, cholesterol and ALP levels showed significant (p<0.05) differences. Significantly (p<0.05) higher titer levels were observed in 100% organic Zn group, 100% and 50% organic Zn, Cu and Mn supplementation groups. The CBH response of all the treated groups showed no significant (p>0.05) difference. At 8 weeks of age, the relative weight of spleen of layer chicks in 100% and 50% organic Zn, Cu and Mn supplementation groups were significantly (p<0.05) higher than all the other treatments. Tibia bone weight (g), tibia calcium (%) and tibia phosphorus (%) varied insignificantly (p>0.05). But significant (p<0.05) difference was observed with respect to the tibia ash content. Except Zn and Mn levels of tibia, all other studied mineral levels of serum and liver did not differ significantly (p>0.05). Faecal excretions of minerals were significantly lower in organic mineral fed groups. Replacement of inorganic Zn, Cu and Mn with corresponding organic minerals improved the body weight, immunity and lower faecal excretion of minerals in chicks.
  Abinash Das , S.K. Mishra , R.K. Swain , P. Swain , K. Dhama , G. Sahoo , N.C. Behura , K. Sethy , B. Chichilichi , T. Behera and S.R. Mishra
  Effect of replacement of inorganic minerals viz., zinc (Zn), copper (Cu) and manganese (Mn) with their corresponding organic minerals (methionine) on growth, bioavailability and immunity was studied in grower birds (9th to 20th week). At 20th week, the body weight of grower birds were significantly (p<0.05) higher in 100% organic Zn group, 100 and 50% organic Zn, Cu and Mn supplemented groups. The cumulative feed consumption and FCR of all the treated groups showed no significant (p>0.05) difference. The serum glucose, after cholesterol SGOT, SGPT and ALP levels differ significantly (p<0.05) of all the treated groups of differed significantly (p<0.05). The CBH response and the antibody titers against SRBC were found to be significantly higher in 100% organic Zn group and 100% Zn, Cu and Mn supplemented groups. Tibia bone weight (g), tibia calcium (%) and tibia phosphorus (%) varied insignificantly (p>0.05). But significant (p<0.05) difference was observed as regard to the tibia ash content. Faecal excretions of Zn, Cu and Mn were significantly lower in organic mineral fed groups. Replacement of inorganic Zn, Cu and Mn with their corresponding organic minerals improved the body weight and immunity, with lower faecal excretion of minerals in grower birds.
  K. Dhama , M. Mahendran and S. Tomar
  The role of migrating wild birds in transmitting diseases of poultry or zoonoses is a contentious issue as the researchers and naturalists stands divided regarding their capability to disperse pathogens over continents. Recently, migratory birds got world wide attention during the bird flu outbreaks, as they were found capable to disseminate the deadly H5N1 avian influenza (bird flu) virus, without themselves getting affected. However, the death of migratory birds due to H5N1, reported from Asia, has fuelled anxiety and concern over the whole issue. Apart from avian influenza, migratory birds are also thought to play role in the transmission of avian viruses like Newcastle disease virus, avian pneumovirus and duck plague virus. Similarly, bacterial pathogens like Chlamydophila psittaci and Pasteurella multocida can be transmitted to domestic poultry via migratory birds. They are also known to spread West Nile virus, equine encephalitis virus, Borrelia burgdorferi and enteropathogens like Campylobacter and Salmonella, which could affect animals as will as human beings. To prevent such etiological agents from entering poultry premises, strict biosecurity and constant surveillance are of paramount importance. Hence, in the scenario of migratory birds contributing significantly to the global spread of infectious diseases, a better understanding of their role in the disease epidemiology has to be gained by implementing superior surveillance and tracking strategies.
  K.A. Naveen , S.D. Singh , J.M. Kataria , R. Barathidasan and K. Dhama
  Newcastle Disease (ND) is a highly contagious infection of poultry which manifest in a wide range of severity from subclinical infection to lethal disease. In the past, a number of Newcastle disease outbreaks in poultry and other bird species have been ascribed to pigeon paramyxovirus type-1 (PPMV-1) infection. The conventional in vivo pathogenicity tests to assess the pathogenicity of PPMV-1 viruses have provided equivocal results. Lately, restriction enzyme analysis technique has been employed for unequivocal identification of individual strains of Newcastle Disease Virus (NDV) in poultry. In this study, sequence analysis of the F1/F2 cleavage site of the F gene of APMV-1 isolated from pigeons in India was attempted for pathotype prediction and determination of molecular epidemiology. Six pigeon origin NDV isolated in India between 1991 and 2001 were selected for this study. A portion of NDV F gene including the cleavage site was amplified by Polymerase Chain Reaction (PCR) and sequenced directly. The total number of nucleotide substitution among all six isolates ranged from 6 to 20; whereas, only four amino acid substitutions were observed. Nucleotides at position 304 and 357 were unique to all the pigeon isolates. The cleavage-activation site (380-397) had no nucleotide substitution and all the six pigeon isolates shared the amino acid sequence 112RRQKRF117 as that of velogenic viruses. The results of this molecular characterization study of Indian PPMV-1 isolates would help design better prevention and control measures for this important pathogen.
  D.B. Barad , B.S. Chandel , A.I. Dadawala , H.C. Chauhan , H.S. Kher , S. Shroff , A.G. Bhagat , S.V. Singh , P.K. Singh , A.V. Singh , J.S. Sohal , S. Gupta , K.K. Chaubey , S. Chakraborty , R. Tiwari , R. Deb and K. Dhama
  Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of chronic enteric granulomatous inflammation in animals and is known as Johne’s Disease (JD) or Paratuberculosis. JD, being spectral in nature, presents variable bacteriological, immunological and pathological spectra leading to variable efficacy of diagnostic methods at different points of time during the course of infection. The present study aimed to estimate the incidence of MAP in two important breeds of goats (Mehsani and Surti) from South Gujarat region of India by applying conventional, molecular and serological methods. A total 219 goats were screened and categorized into Group-I (123 Mehsani goats), Group-II (76 Surti goats) and Group-III, (20 Non-descript goats). Percent positivity by faecal smear examination, delayed type hypersensitivity (DTH), agar gel immunodiffusion (AGID), IS900 polymerase chain reaction) (PCR) and indigenous enzyme linked immunosorbent assay (ELISA) kit was 9.2 (7/76), 21.9 (27/123), 10.9 (24/219), 12.5 (5/40) and 43.3% (95/219), respectively. Of the 123 goats of Group-I, 27 (21.9%) were positive in DTH test. Of the 5 faecal positive goats which also showed clinical signs, 2 (3.5%) goats died during study were negative by Johnin test. Similar to these findings, sensitivity of Johnin test in goats ranged between 18-30% with least specificity in both preclinical and advanced stage of disease. Of 34 cases of caprine paratuberculosis, 73.5% goats were positive for Johnin test. In the present study, out of the 5 infected goats, 3 (60%) were positive in Johnin test. Rectal pinch smear examination was carried out in 27 DTH positive goats and all smears were negative for the presence of acid fast bacilli. Screening tests (Indigenous ELISA and DTH) showed very high incidence of MAP infection in the goat population. The utility of multiple diagnostic tests is suggested for confirmatory detection and epidemiological diseases investigations of MAP in animals.
  A. Kumar , S.V. Singh , A.K. Srivastava , N.K. Gangwar , P.K. Singh , S. Gupta , K.K. Chaubey , R. Tiwari , S. Chakraborty and K. Dhama
  Johne’s Disease (JD), caused by Mycobacterium avium subspecies paratuberculosis, is endemic in domestic animals and adversely affects dairy industry worldwide. In the present study, efficacies of ‘Indigenous’and commercial (Gudair, Spain) vaccines were evaluated for control of JD in experimentally challenged goats. Goats were grouped into Sham-immunized, Indigenous and Gudair vaccine groups. Vaccinated kids were challenged at 50 and 270 Days Post Vaccination (DPV), with 3×109 and 5×109 ‘Indian Bison Type strain ‘S 5’, respectively and sacrificed at 150 and 450 DPV after 1st and 2nd challenge, respectively. Vaccines were evaluated for improvements in physical condition (diarrhea, weakness, body coat color), clinical symptoms (shedding of bacilli, mortality, morbidity), immune responses (cell-mediated and humoral), pathology (gross and microscopic lesions) and production status (body weights, growth rates). Vaccinated goats gained higher body weights vis a vis sham-immunized. Mortality was higher in sham-immunized. Cell Mediated Immunity (CMI) response increased at 30 DPV and showed down regulation from 90 DPV onwards in vaccinated goats. Significant increase in humoral immune response was observed in vaccinated goats at 180 DPV and maintained till 450 DPV. Microscopical examination at 180 DPV showed reduced shedding in vaccinated groups, At 200 DPV, group 1 goats showed thickening of small intestine with corrugations specifically at ileocaecal junction, catarrhal enteritis with infiltration of mononuclear cells and epitheloid cells. In vaccinated groups, there were focal thickening of intestines at 450 DPV with lesions of chronic catarrhal enteritis and presence of lymphocyte, plasma cells and macrophages cells with a few epitheloid cells. Monitoring of MAP DNA in the blood of experimental goats of all the groups was done by testing of blood samples by Polymerase Chain Reaction (PCR) and the vaccinated groups of goats revealed MAP bacilli free status upto 300 DPV. Both the vaccines provided protection after challenge I, but since indigenous vaccine also protected goats after challenge II, was therefore superior. In conclusion, the indigenous vaccine must be exploited for its full potential for effective prevention and control of this economically important disease having public health concerns.
  V. Gowthaman , S.D. Singh , K. Dhama , R. Barathidasan , M. Asok Kumar , P.A. Desingu , N.K. Mahajan and M.A. Ramakrishnan
  Adenoviruses have been isolated from both clinically healthy and diseased birds worldwide. The pathogenic role of most of the FAdVs is still questionable. They can quickly take on the role of opportunistic pathogens when additional factors, particularly concurrent infections, adversely affect the health of the avian host. Immnosuppressing agents especially chicken infectious anemia and infectious bursal disease viruses are known to enhance the pathogenicity of FAdVs upon coinfection. The aim of the present study was to screen for the involvement of FAdV in poultry flocks affected with respiratory disease complex by RT-PCR. The samples were also screened by RT-PCR/PCR for other respiratory pathogens. Thirty two commercial poultry flocks with the history of respiratory disease complex from various parts of India. FAdV nucleic acid could be detected in tissue samples of 13 out of 34 farms investigated. Out of 13 FAdV positive farms, FAdV and CIAV were alone detected in 4/13 (31%) whereas, in other farms more than two respiratory pathogens were detected together. CIAV was detected in all the farms (34/34) investigated. Eosinophilic intranuclear inclusion bodies were noticed in FAdV infected laryngeal and tracheal epithelium under light microscopy. The findings of the study assert that FAdV can play the role of primary respiratory pathogen in immunocompromised birds and also in the presence of other respiratory pathogens.
  Mahima , Anu Rahal , Rajib Deb , Shyma K. Latheef , Hari Abdul Samad , Ruchi Tiwari , Amit Kumar Verma , Amit Kumar and K. Dhama
  Herbs/Botanical plants are considered as God’s gift to human beings in the form of natural medicines, like the one well known “Sanjeevani booti” described in Hindu Mythology. The traditional and ethno-veterinary practices have been in use for centuries, transferring the knowledge from generation to generation and they are accessible, easy to prepare and administer, with little or no cost at all. Even though the modern developments in therapeutic field brought about a rapid decline in traditional medicine, the plant-based remedies are still having a crucial role as potential source of therapeutic aids in health systems all over the world for both humans and animals. Among the 21,000 medicinal plants listed by the World Health Organization (WHO), 2500 species are native to India, which stands first in the production of medicinal herbs. This innumerable treasure of medicinal herbs brings India the distinction of ‘the botanical garden of the world’. Nowadays immune-based therapies are gaining more importance than monovalent approaches which are having limited benefits. Apart from the actions like treating diseases, control of ecto- and endo-parasites, fertility enhancement, bone setting and poor mothering management, an array of herbal medicines have been reported which are having immunomodulatory effects like modulation of cytokine secretion, histamine release, immunoglobulin secretion, class switching, cellular co-receptor expression, lymphocyte expression, phagocytosis and so on. The present article describes in brief few of these important ones viz., ashwagandha, amla, tulsi, arjuna, aloe vera, garlic, turmeric, ginger, shatavari, neem, guduchi, kiwifruit, tut, kamala, palashlata, kokilaksha etc. being used for human and animal health benefits.
  Amit Kumar Verma , Amit Kumar , K. Dhama , Rajib Deb , Anu Rahal , Mahima and Sandip Chakraborty
  Leptospirosis, caused by pathogenic spirochetes of the genus Leptospira, affects both humans and animals and is among the most common but neglected direct zoonotic disease in the world, particularly in untreated or undiagnosed animals as well as humans. Now, it has been considered as a re-emerging disease causing global health problem due to its increasing incidences in developing as well as developed nations. It is a multisystemic disease leading to death. Diagnostic tests of importance are Latex Agglutination Test (LAT), lateral flow and immunoglobulin M (IgM) based ELISA, PCR based assays, Multiple-microscopic Agglutination Test (MAT), Loop-mediated Isothermal Amplification (LAMP) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Molecular tools like PCR-RFLP, real-time PCR, multiplex PCR, qPCR and immunocapture PCR have all been found useful for rapid and confirmatory detection and differentiation of pathogenic and non-pathogenic leptospires. Inactivated/killed and attenuated vaccines are always attempted, since the beginning of vaccine and vaccination story, against all emerging pathogens with mixed success stories. The advanced tools and techniques like recombinant DNA technology, reverse genetics, DNA vaccination, molecular genetics and proteomics approaches are being explored for search of novel antigens, proteins and genes as potential candidates to discover safer, efficient and better vaccines for leptospirosis. The present review highlights the leptospirosis, susceptible population, disease transmission and epidemiology, treatment, trends and advances in diagnosis, vaccines and vaccination strategies in humans and animals with a view to combat this organism having public health significance.
  K. Dhama , S. Rajagunalan , S. Chakraborty , A.K. Verma , A. Kumar , R. Tiwari and S. Kapoor
  The term food borne diseases or food-borne illnesses or more commonly food poisoning are used to denote gastrointestinal complications that occur following recent consumption of a particular food or drink. Millions of people suffer worldwide every year and the situation is quiet grave in developing nations creating social and economic strain. The food borne pathogens include various bacteria viz., Salmonella, Campylobacter, Escherichia coli, Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus, Arcobacter, Clostridium perfringens, Cl. botulinum and Bacillus cereus and helminths viz., Taenia. They also include protozoa viz., Trichinella, Sarcocystis, Toxoplasma gondii and Cryptosporidium parvum. The zoonotic potential and the ability to elaborate toxins by many of the microbes causing fatal intoxication are sufficient to understand the seriousness of the situation. The viral agents being host specific their transmission to humans through food of animal origin is not yet confirmed although these animal viruses are similar to that of viruses infecting human. Food-borne bacteria; protozoa and helminthes have complex distribution pattern in the environment and inside the host system. This along with complexity of the maintenance chain and life cycle (of parasites) has made it difficult for epidemiologist and diagnostician to undertake any immediate safety measures against them. Serological and molecular diagnostic tests viz. ELISA, Latex agglutination test, Lateral flow assays, Immunomagnetic separation assays, molecular assays viz. Polymerase Chain Reaction (PCR), multiplex PCR, immuno-PCR, Realtime PCR, Random Amplified Polymorphic DNA (RAPD)-PCR, DNA microarrays and probes are widely used. Along with these LAMP assays, Capillary Electrophoresis-Single Strand Confirmation polymorphism (CE-SSCP); Flow cytometry, FISH, Biosensors, Direct epifluorescent filter technique, nanotechnology based methods and sophisticated tools (ultrasonography, magnetic resonance imaging and chlonangio-pancreatography) have aided in the diagnosis greatly. Most of the food-borne illnesses are self-limiting but in many instances antibiotics are recommended. With the increased drug resistance however use of chicken immunoglobulin, bacteriophage therapy, probiotics and herbs are gaining much importance these days. Adoption of proper prevention and control measures (including cooking procedures; hygiene, strict adherence to HACCP principles, public awareness and disease surveillance and monitoring) are the need of hour. All these have been discussed vividly in this review to help epidemiologists, diagnosticians, clinicians and above all common people so as to enable them avoid negligence regarding such serious issue.
  A. Hansa , R.B. Rai , K. Dhama , M.Y. Wani , M. Saminathan and G.J. Ranganath
  Bovine Coronavirus (BCoV) is widespread both in dairy and beef cattle throughout the world. The virus is one of the largest RNA virus and has specific tropism for intestinal and pulmonary epithelial cells. It is responsible for huge economic losses by causing winter dysentery in adult dairy cattle and respiratory and intestinal tract infections leading to pneumo-enteritis in young calves. Isolation of BCoV has been reported to be difficult. Studies regarding epidemiology, virus isolation and molecular detection from India are very few. In the present study Vero cell line was used for isolation of the BCoV from Enzyme Linked Immunosorbent Assay (ELISA) positive samples. Direct florescent antibody technique (dFAT) and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to confirm the isolated virus strains at antigenic and genomic levels, respectively. Out of the 15 positive fecal samples, virus from only seven was able to infect vero cell line. Subsequently BCoV got adapted to the vero cell line upto three passages, which was confirmed both at genomic and antigenic levels by dFAT and RT-PCR testing. It can be concluded that vero cell line can be used for isolation of BCoV, however due to the enormous stain diversity of the virus it is possible that many stains can’t grow and get adapt in this cell line. Further studies are required for isolation of different viral strains, finding the susceptible cell lines and also to confirm the variations among the BCoV isolates at antigenic/genomic levels.
  K. Dhama , S. Kapoor , R.V.S. Pawaiya , S. Chakraborty , R. Tiwari and A.K. Verma
  A fascinating and important arbovirus is Ross River Virus (RRV) which is endemic and epizootic in nature in certain parts of the world. RRV is a member of the genus Alphavirus within the Semliki Forest complex of the family Togaviridae, which also includes the Getah virus. The virus is responsible for causing disease both in humans as well as horses. Mosquito species (Aedes camptorhynchus and Aedes vigilax; Culex annulirostris) are the most important vector for this virus. In places of low temperature as well as low rainfall or where there is lack of habitat of mosquito there is also limitation in the transmission of the virus. Such probability is higher especially in temperate regions bordering endemic regions having sub-tropical climate. There is involvement of articular as well as non-articular cells in the replication of RRV. Levels of pro-inflammatory factors viz., tumor necrosis factor-alpha (TNF-α); interferon-gamma (IFN-γ); and macrophage chemo-attractant protein-1 (MAC-1) during disease pathogenesis have been found to be reduced. Reverse transcription-polymerase chain reaction (RT-PCR) is the most advanced molecular diagnostic tool along with epitope-blocking enzyme-linked immunosorbent assay (ELISA) for detecting RRV infection. Treatment for RRV infection is only supportive. Vaccination is not a fruitful approach. Precise data collection will help the researchers to understand the RRV disease dynamics and thereby designing effective prevention and control strategy. Advances in diagnosis, vaccine development and emerging/novel therapeutic regimens need to be explored to their full potential to tackle RRV infection and the disease it causes.
  Amit Kumar , Amit Kumar Verma , Anu Rahal , Pramod Kumar Panwar and K. Dhama
  Adjuvants are used as a carrier of antigen in modern vaccine therapy. These are heterogeneous compounds which are administered with antigens to elicit better immune response against co-administered antigens by stimulating the immune responses. Ideally, an adjuvant should not be mutagenic, carcinogenic and teratogenic and it should not produce any autoimmune disease. However, many of the adjuvants used in vaccine preparation have one or another side effect. The present paper describes in brief the history, development and recent trends in the adjuvant of vaccine.
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