China-Pakistan Economic Corridor: A Case Study Of Internal Security Challenge Faced by Pakistan

The strongest and most crucial strategic and economic links in the south Asian region are the friendships between Pakistan and China. China and Pakistan assert that the people who live on either side of the Himalayas have interacted with one another in a trustworthy and sincere manner from ancient times. Chinese investment in the CPEC represents the greatest sum (62 billion dollars) ever between two friendly nations. China and Pakistan are dealing with numerous internal and external security challenges as a result of their strong friendship and significant investment in this project. Both nations, such as India and Afghanistan, are posing security problems to Pakistan, which has several security issues. The USA, India, the UK, and Israel are also causing strife and tension in Pakistan. These nations are also causing internal problems by sponsoring terrorist organizations like the Taliban and ISIS as well as nationalist organizations like the Baloch, the Lashkar-e-Jhangvi, and Sipah Muhammad. The challenges to Pakistan's internal security are discussed in this study.

Production and Immunological Evaluation of Polyvalent Vaccine of Avian Influenza Aiv-H9 and Fowl Cholera Pasteurella Multocida

Infectious diseases particularly fowl cholera (FC) and avian influenza (AI) are challenge for birds, causes severe economic loss and hampers to the development of poultry industry in the developing countries including Pakistan. Vaccination programs using monovalent vaccines against the diseases are effective but the repeated vaccination induces stress to the animals and increase cost of vaccination for the farmer. Owing to significant role of hemagglutinin (HA) and neuraminidase (NA) gene in antigenic drift, AIV field isolates leads to escape the immunity provided by vaccines. In this study, the genetic and antigenic nature was explored in currently prevailing H9N2 viruses in Pakistan during year 2017. The efficacy of bivalent AI+FC vaccine was compared to monovalent AI and FC vaccines. H9N2 AIV samples were collected from affected poultry flocks from different areas of the Punjab and were serologically and molecularly confirmed. Positive isolates were propagated on chicken fibroblast cell line. Direct sequencing of HA and NA gene was performed and phylogenetic relationship of identified strains was evaluated. The distribution of mutations was analyzed on different epitopes of HA antigen. To check the antigenic effect of potential predicted substitutions at different positions of HA surface glycoprotein, protein-peptide modelling was done by GlaxayWEB. Previously isolated and characterized AIV subtype “H9N2” and Pasteurella multocida were used as antigen in the vaccine production. Formalin inactivated alum and oil adjuvated AI, FC and AI+FC vaccines were prepared and injected in experimental poultry birds at 0 and 14th day. The serum samples were collected on 0, 14, 21, 48 and 60 days post-priming. Antibody titer was measured by Hemagglutinin Inhibition (HI) for AI “H9N2” virus and enzyme-linked immunosorbent assay (ELISA) was performed for lipopolysaccharide (LPS) of P. multocida. The results showed that H9N2 virus had 99% homology with the earlier reported AIV lineages of Iran, India, Saudi Arabia and Pakistan while their phylogenetic examination showed that the strains of this study belonged to the Pakistani H9N2 viruses isolated during years 2010-2015. The amino acid analysis of isolates (H9N2) showed substitution of A to T at position 198 of HA receptor-binding motif. The in-silico predictive changes in HA’s antigenic sites R180A, T163A and N201A changed the 3D-structure of protein-peptide binding site. The birds exhibited the classical pattern of immune response in multivalent formulation (both oil and alum based) with achievement of peak antibody titers (HI titer 256 and 128; ELISA titer 2.752 and 1.805) at 48th day post vaccination. The HI and ELISA titers in combined vaccine were significantly higher (P < 0.05) than corresponding monovalent formulation. Oil based combined vaccine showed better immune response due to prolonged immunity. In conclusion, our findings demonstrates that present H9N2 AI isolates circulating in poultry flocks were related to previous strains of H9N2 in Pakistan. The change of Amino acid at antigenic sites 163, 180 and 201 play a vital role in determination of the antigen binding to antibody, which may lead to escape immunity provided by vaccine. From immunological point of view, AI vaccine can be combined with FC vaccine and gave better immune response than alone one. Being gram negative, the LPS of P. multocida may acts as a factor contributing towards relatively higher antibody titer in comparison with the respective monovalent vaccine. This combined vaccine may also be helpful to reduce burden as repeated vaccination itself is a stress for poultry birds. It would also be helpful for poultry breeders in reducing the cost as well as frequency of vaccination, leading to less stress and higher immune response in birds. Moreover, oil based combined vaccine is better choice for prolonged titer than alum adjuvanted combined vaccine.