Cytological and immunocytological detection and differentiation of Marek's disease and lymphoid leucosis in poultry.

Marek's disease (MD) and lymphoid leucosis (LL) are the major diseases causing lymphoid tumors in chickens accounting for high economical losses. Gross examination could not yield definite diagnosis owing to their similar presentation of lesions. Thus present work was aimed for diagnosis and differentiation of MD and LL by utilizing simple cytology and novel immunocytology techniques. Cytological examination was carried out on slides with tumor touch imprints stained by simple Giemsa staining. The diagnosis was mainly achieved based on morphology of cell population. In the present study, out of a total of 595 cases examined, 502 cases had pleomorphic lymphocytic cell population suggestive of MD and 53 cases had uniform lymphocytic/lymphoblast cell population suggestive of LL, while the rest 40 cases remained inconclusive. A definitive diagnosis was achieved after performing immunocytology using specific antibodies that revealed 518 cases had reactivity for Meq oncoprotein specific for MD and 77 cases showed immunoreactivity for IgM in transformed B-cells confirming LL. The technique of immunocytology which has been useful for detecting human viral pathogens and MD in poultry has been applied for the first time as a novel, simple, rapid and inexpensive technique that could be used as an alternate test to effectively detect and differentiate MD and LL in poultry.

Immunomodulatory and prophylactic efficacy of herbal extracts against experimentally induced chicken infectious anaemia in chicks: assessing the viral load and cell mediated immunity.

Chicken infectious anaemia virus (CIAV) is an economically important and a highly immunosuppressive virus affecting poultry industry worldwide. In this study we assessed the immunomodulatory effects of four herbal preparations namely Withania somnifera, Tinospora cordifolia, Azadirachta indica and E Care Se Herbal in resisting the viral multiplication and immunosuppression inflicted by CIAV in chicks. Day-old chicks (n = 90) were randomly and equally divided into six groups (Groups A-F). Groups A-D were administered with purified extracts of W. somnifera, T. cordifolia, A. indica and E Care Se Herbal, respectively followed by the evaluation of viral load in lymphoid organs by quantitative real-time PCR and cell mediated immune response by flow cytometric analysis of CD4+ and CD8+ T cells. Groups A-D were found to resist CIAV multiplication and pathogenesis with significant reduction of viral load compared with the infected control (P < 0.05). Group A-C chicks showed significantly higher (P < 0.05) CD4+ and CD8+ T cell counts compared to control birds while of E Care Se Herb had minimal effect on T cell count. The findings suggested that the herbal preparations used during the study were effective as both prophylactic and immunomodulatory agents and thus have potential of being used against CIAV induced immunosuppression in poultry.

Duck virus enteritis (duck plague) - a comprehensive update.

Duck virus enteritis (DVE), also called duck plague, is one of the major contagious and fatal diseases of ducks, geese and swan. It is caused by duck enteritis virus (DEV)/Anatid herpesvirus-1 of the genus Mardivirus, family Herpesviridae, and subfamily Alpha-herpesvirinae. Of note, DVE has worldwide distribution, wherein migratory waterfowl plays a crucial role in its transmission within and between continents. Furthermore, horizontal and/ or vertical transmission plays a significant role in disease spread through oral-fecal discharges. Either of sexes from varying age groups of ducks is vulnerable to DVE. The disease is characterized by sudden death, vascular damage and subsequent internal hemorrhage, lesions in lymphoid organs, digestive mucosal eruptions, severe diarrhea and degenerative lesions in parenchymatous organs. Huge economic losses are connected with acute nature of the disease, increased morbidity and mortality (5%-100%), condemnations of carcasses, decreased egg production and hatchability. Although clinical manifestations and histopathology can provide preliminary diagnosis, the confirmatory diagnosis involves virus isolation and detection using serological and molecular tests. For prophylaxis, both live-attenuated and killed vaccines are being used in broiler and breeder ducks above 2 weeks of age. Since DEV is capable of becoming latent as well as shed intermittently, recombinant subunit and DNA vaccines either alone or in combination (polyvalent) are being targeted for its benign prevention. This review describes DEV, epidemiology, transmission, the disease (DVE), pathogenesis, and advances in diagnosis, vaccination and antiviral agents/therapies along with appropriate prevention and control strategies.

Haemorrhagic enteritis of turkeys - current knowledge.

Haemorrhagic enteritis virus (HEV), an adenovirus associated with acute haemorrhagic gastro-intestinal disease of 6-11-week old turkeys predominantly hampers both humoral and cellular immunity. Affected birds are more prone to secondary complications (e.g. colibacillosis and clostridiosis) and failure to mount an effective vaccine-induced immune response. HEV belongs to the new genus Siadenovirus. Feco-oral transmission is the main route of entry of the virus and it mainly colonizes bursa, intestine and spleen. Both naturally occurring virulent and avirulent strains of HEVs are serologically indistinguishable. Recent findings revealed that ORF1, E3 and fib genes are the key factors affecting virulence. The adoption of suitable diagnostic tools, proper vaccination and biosecurity measures have restrained the occurrence of disease epidemics. For diagnostic purposes, the best source of HEV is either intestinal contents or samples from spleen. For rapid detection highly sensitive and specific tests such as quantitative real-time PCR based on Taq man probe has been designed. Avirulent strains of HEV or MSDV can be effectively used as live vaccines. Novel vaccines include recombinant hexon protein-based subunit vaccines or recombinant virus-vectored vaccines using fowl poxvirus (FPV) expressing the native hexon of HEV. Notably, subunit vaccines and recombinant virus vectored vaccines altogether offer high protection against challenge or field viruses. Herein, we converse a comprehensive analysis of the HEV genetics, disease pathobiology, advancements in diagnosis and vaccination along with appropriate prevention and control strategies.

Evaluating a murine model of endometritis using uterine isolates of Escherichia coli from postpartum buffalo.

Ascending infection of the uterus with Gram-negative bacteria is responsible for postpartum endometritis in cattle and buffalo and can adversely affect fertility. Development of a laboratory animal model for bovine endometritis would facilitate the understanding of the pathogenesis as it is difficult to conduct controlled experimentation in the native host. In the present study, 30 virgin Swiss Albino mice (5-8 weeks old) were used to evaluate the pathogenic potential of Escherichia coli, isolated from the normally calved postpartum buffalo to induce endometritis. Mice in the diestrus phase of the estrous cycle were randomly allotted to one of the following four intravaginal inoculation (100 µL) treatments: EG (experimental group)-1: sterile normal saline; EG-2, -3 and -4: E. coli@ 1.5 × 104, 105 and 106 CFU/ml, respectively. The animals were then scarified 36 h post-inoculation to study gross and microscopical lesions. Gross changes were confined to EG-4. Acute endometritis was recorded in 50% of the EG-3 and 66.7% of the EG-4. The rate of acute endometritis development was significantly higher in EG-4 (P<0.05) as compared to the other groups. The present study demonstrated that the animal model for bubaline endometritis can be developed in mice by intravaginal inoculation of E.coli@ 1.5 × 106 CFU/ml at diestrus. Ease of intravaginal inoculation, apparent absence of systemic involvement and high infective rate are the advantages of the model over other studies.

A novel recombinant Meq protein based dot-ELISA for rapid and confirmatory diagnosis of Marek's disease induced lymphoma in poultry.

Marek's disease (MD), is an economically important virus disease of poultry throughout the world. In this study, we for the first time reports development of a novel dot enzyme-linked immunosorbent assay (dot-ELISA) for the confirmatory diagnosis of lymphoma caused by Marek's Disease Virus (MDV). Suspected lymphoma tissue extracts from the diseased birds were used for the Meq oncoprotein antigen detection, which is expressed specifically in MDV lymphomas. Recombinant Meq oncoprotein was expressed using Expresso™ Rhamnose Sumo Cloning and Expression system and the hyperimmune serum was raised against it, which was used later while developing dot-ELISA. The dot-ELISA exhibited higher specificity (92%) in diagnosing MD lymphomas as compared to conventional PCR (40%), where later assay is unable to differentiate disease development (lymphoma) and/or infection. The developed dot-ELISA proved to be a specific, rapid and inexpensive technique detecting MDV lymphomas in poultry. Of the note, this new assay could be opted as a valuable diagnostic tool in the resource poor countries andcould further be used to differentiate from other tumor causing viruses in poultry.