PRIMARY CELL CULTURE OF AEDES ALBOPICTUS MIDGUT CELLS: A PROSPECTIVE MODEL FOR IN VITRO STUDY OF ARBOVIRUSES

  Objective: Midgut cells play a key role in the propagation of mosquito borne Arboviruses. The existing mosquito cell lines for studying viral pathogenesis are derived either from larvae or from eggs since there is no cell line available from the mosquito midgut. Therefore, to delineate the in situ viral interaction which naturally occurs within the mosquito midgut and represent cellular pathogenesis in human beings, the present work was aimed to develop a primary cell line from the midgut cells of Aedes albopictus.Methods: The midgut cells of A. albopictus were collected, cultured and incubated at 28°C to study the growth after every 24 hrs for 7 days.Result: The primary cell culture showed an increasing growth pattern of columnar cells up to 48 hrs followed by decrease in cell population afterward. However, the number of stem cells increased significantly throughout the study period, and their population outnumbered the columnar cells after 72 hrs. There was no significant change of goblet cells and regenerating cells which were scanty in number throughout the experiment.Conclusion: The present method will help to develop the individual cell lines from mosquito midgut and study the host pathogen interaction in arboviral diseases in future

Seroprevalence of viral and bacterial diseases among the bovines in Himachal Pradesh, India

Aim: The study was designed to measure the seroprevalence of viral and bacterial diseases: Infectious bovine rhinotracheitis, bovine viral diarrhea, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, brucellosis, and paratuberculosis among bovine of Himachal Pradesh during the year 2013-2015. Materials and Methods: The serum samples were collected from seven districts of state, namely, Bilaspur, Kangra, Kinnaur, Lahul and Spiti, Mandi, Sirmour, and Solan. The samples were screened using indirect ELISA kits to measure the seroprevalence of viral and bacterial diseases. Results: The overall seroprevalence of infectious bovine rhinotracheitis was 24.24%, bovine viral diarrhea 1.52%, bovine leukemia 9.09%, bovine parainfluenza 57.58%, bovine respiratory syncytial disease 50%, brucellosis 19.69%, and paratuberculosis 9.09% in Himachal Pradesh. The seroprevalence of bovine rhinotracheitis, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, and paratuberculosis in the state varied significantly (p0.01). Multiple seropositivity has been observed in this study. Bovine parainfluenza virus 3 was observed commonly in mixed infection with almost all viruses and bacteria under study. Conclusion: The viral and bacterial diseases are prevalent in the seven districts of Himachal Pradesh investigated in the study. Therefore, appropriate management practices and routine vaccination programs should be adopted to reduce the prevalence of these diseases

Changes in viral load in different organs of Japanese Encephalitis virus-infected chick embryo under the influence of Belladonna 200C

Background: Japanese encephalitis(JE) is highly prevalent in many states of India. Belladonna 200C is widely used in the prevention and treatment of JE.The effect of Belladonna 200C in virus replication inside different tissues has not been studied. Objective: To study the effect of Belladonna 200C in virus replication inside different tissues utilising chick embryo model. Materials and Methods: Twelve-day-old fertilised eggs of Black Australorp were inoculated with JE via chorioallantoic membrane (CAM) route in different experimental sets: infection, Belladonna 200C treated and vehicle control, keeping matched blank sets. All experimental sets were incubated for 48 hours. After incubation, viable eggs were sacrificed humanly and different tissues were observed and collected for viral load determination by real-time-polymerase chain reaction (PCR). Results: The control group showed visible pocks over the CAM; brains were liquefied due to haemorrhagic liquefactive necrosis and white patches were found over the liver. However, the medicine-treated group was apparently normal; there were no visible changes in the brain and the liver was healthy like control. Real-time-PCR results showed high viral load in CAM and brain with absence of viral RNA in liver of the virus-infected group. Pre-treatment with Belladonna 200C significantly reduced the overall load (P < 0.05) in CAM and brain which correlated with the morbid pathological changes of the organs. Conclusion: Although Belladonna 200C did not completely inhibit JE viral replication in the brain, it reduced the severity of JE by diminishing the viral loads in this tissue