Ali Saeed

Detection and Phylogenetic Analysis of Peste des Petits Ruminants Virus Isolated from Outbreaks in Punjab, Pakistan

Peste des Petits Ruminants (PPR) is an important viral disease of small ruminants and is endemic in Pakistan. In the following study, samples from two outbreaks of PPR in goats have been subjected to laboratory investigations. The Peste des Petits Ruminants virus (PPRV) genome was detected using both conventional and real-time PCR. Genetic characterization of the local PPRV field isolates was conducted by sequencing 322u2003bp of the fusion (F) gene and 255 bp of the nucleoprotein (N) gene. The phylogenetic tree based on the F gene clustered samples from both outbreaks into lineage 4 along with other Asian isolates, specifically into subcluster 1 along with isolates from Middle East. Analysis of N gene revealed a different pattern. In this case, the Pakistani samples clustered with Chinese, Tajikistani and Iranian isolates, which probably represents the true geographical pattern of virus circulation. This is the first report presenting the phylogenetic tree based on N gene as well as performing a parallel comparison of the trees of F and N gene together from Pakistani isolates. The results of this study shed light on the PPRV population in Pakistan and emphasize the importance of using molecular methods to understand the epidemiology. Such understanding is essential in any efforts to control the number and impact of outbreaks that are occurring in endemic countries such as Pakistan, especially in the current scenario where OIE and FAO are eager to control and subsequently eradicate PPR from the globe, as has been achieved for Rinderpest.

Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab.

OBJECTIVEnTo determine the presence of Babesia bovis (B. bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals.nnnMETHODSnBlood samples were collected from 144 large ruminants, including 105 cattle and 39 buffaloes, from six districts in southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar and Vehari. Data on the characteristics of animals and herds were collected through questionnaires. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B. bovis on the blood and serological profile of infected animals.nnnRESULTSn27 out of 144 animals, from 5 out of 6 sampling districts, produced the 541-bp fragment specific for B. bovis. Age of animals (P=0.02), presence of ticks on animals (P=0.04) and presence of ticks on dogs associated with herds (P=0.5) were among the major risk factors involved in the spread of bovine babesiosis in the study area. ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle.nnnCONCLUSIONSn: This study has reported for the first time the presence of B. bovis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output.

Molecular Characterization of Peste des Petits Ruminants Viruses From Outbreaks Caused by Unrestricted Movements of Small Ruminants in Pakistan

Peste des petits ruminants (PPR) is an endemic disease of small ruminants, and vaccination has been the method of control but outbreaks are continuously occurring in Pakistan. The following study presents a detailed investigation of an outbreak, suspected to be PPR, probably introduced by PPRV-infected sheep and goats from Sindh Province (north-west) to Punjab Province (central) of Pakistan during the flood relief campaign in 2011. A total of 70 serum samples from 28 different flocks were tested with competitive ELISA (H antibodies), which detected 24 (34.2%) samples positive for PPRV antibodies. Nasal swabs and faeces were tested with immunocapture ELISA (N antigen), which detected 18 (25.7%) samples positive for PPRV antigen. The RNA detected positive (nxa0=xa028, 40%) using real-time PCR was subjected to conventional PCR for the amplification of the fusion and nucleoprotein genes. Sequencing of both genes and subsequent phylogenetic analysis indicated the grouping of all the sequences to be in lineage IV along with other Asian isolates of PPRV. However, sequences of both genes were divided into two groups within lineage IV. One group of viruses clustered with previously characterized Pakistani isolates, whereas the other group was distinctly clustered with isolates from the Middle East or India. The sequence identity indicated the introduction of at least one population of PPRV from a different source and circulation in the local flocks of small ruminants, which emphasized the need to obtain health clearance certificate before movement of animals. The results of this study provide baseline data for the genetic characterization of different PPRV populations in Pakistan.

Foot-and-mouth disease: overview of motives of disease spread and efficacy of available vaccines

Control and prevention of foot and mouth disease (FMD) by vaccination remains unsatisfactory in endemic countries. Indeed, consistent and new FMD epidemics in previously disease-free countries have precipitated the need for a worldwide control strategy. Outbreaks in vaccinated animals require that a new and safe vaccine be developed against foot and mouth virus (FMDV). FMDV can be eradicated worldwide based on previous scientific information about its spread using existing and modern control strategies.

Disturbed Vitamin A Metabolism in Non-Alcoholic Fatty Liver Disease (NAFLD)

Vitamin A is required for important physiological processes, including embryogenesis, vision, cell proliferation and differentiation, immune regulation, and glucose and lipid metabolism. Many of vitamin A’s functions are executed through retinoic acids that activate transcriptional networks controlled by retinoic acid receptors (RARs) and retinoid X receptors (RXRs).The liver plays a central role in vitamin A metabolism: (1) it produces bile supporting efficient intestinal absorption of fat-soluble nutrients like vitamin A; (2) it produces retinol binding protein 4 (RBP4) that distributes vitamin A, as retinol, to peripheral tissues; and (3) it harbors the largest body supply of vitamin A, mostly as retinyl esters, in hepatic stellate cells (HSCs). In times of inadequate dietary intake, the liver maintains stable circulating retinol levels of approximately 2 μmol/L, sufficient to provide the body with this vitamin for months. Liver diseases, in particular those leading to fibrosis and cirrhosis, are associated with impaired vitamin A homeostasis and may lead to vitamin A deficiency. Liver injury triggers HSCs to transdifferentiate to myofibroblasts that produce excessive amounts of extracellular matrix, leading to fibrosis. HSCs lose the retinyl ester stores in this process, ultimately leading to vitamin A deficiency. Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome and is a spectrum of conditions ranging from benign hepatic steatosis to non-alcoholic steatohepatitis (NASH); it may progress to cirrhosis and liver cancer. NASH is projected to be the main cause of liver failure in the near future. Retinoic acids are key regulators of glucose and lipid metabolism in the liver and adipose tissue, but it is unknown whether impaired vitamin A homeostasis contributes to or suppresses the development of NAFLD. A genetic variant of patatin-like phospholipase domain-containing 3 (PNPLA3-I148M) is the most prominent heritable factor associated with NAFLD. Interestingly, PNPLA3 harbors retinyl ester hydrolase activity and PNPLA3-I148M is associated with low serum retinol level, but enhanced retinyl esters in the liver of NAFLD patients. Low circulating retinol in NAFLD may therefore not reflect true “vitamin A deficiency”, but rather disturbed vitamin A metabolism. Here, we summarize current knowledge about vitamin A metabolism in NAFLD and its putative role in the progression of liver disease, as well as the therapeutic potential of vitamin A metabolites.

The interrelationship between bile acid and vitamin A homeostasis

Vitamin A is a fat-soluble vitamin important for vision, reproduction, embryonic development, cell differentiation, epithelial barrier function and adequate immune responses. Efficient absorption of dietary vitamin A depends on the fat-solubilizing properties of bile acids. Bile acids are synthesized in the liver and maintained in an enterohepatic circulation. The liver is also the main storage site for vitamin A in the mammalian body, where an intimate collaboration between hepatocytes and hepatic stellate cells leads to the accumulation of retinyl esters in large cytoplasmic lipid droplet hepatic stellate cells. Chronic liver diseases are often characterized by disturbed bile acid and vitamin A homeostasis, where bile production is impaired and hepatic stellate cells lose their vitamin A in a transdifferentiation process to myofibroblasts, cells that produce excessive extracellular matrix proteins leading to fibrosis. Chronic liver diseases thus may lead to vitamin A deficiency. Recent data reveal an intricate crosstalk between vitamin A metabolites and bile acids, in part via the Retinoic Acid Receptor (RAR), Retinoid X Receptor (RXR) and the Farnesoid X Receptor (FXR), in maintaining vitamin A and bile acid homeostasis. Here, we provide an overview of the various levels of communication between vitamin A metabolites and bile acids and its relevance for the treatment of chronic liver diseases.

Pathogenicity of avian influenza virus H5N1 2007 isolates from Pakistan

Abstract Objective To assess the pathogenicity and to categorize the high pathogenic avian influenze (HPAI) H5N1 of four isolates from different outbreaks during 2007. Methods Primary assays of pathogenicity test were used such as mean death time (MDT), intracerebral pathogenicity index (ICPI) and intravenous pathogenicity index. Results The MDT of isolate 1, 2, 3 and 4 was 35 h, 36 h, 38 h and 35 h respectively in ten-day old chicken embryonated eggs. The ICPI in one day old chick for all isolates were more than 0.7 and the intravenous pathogenicity indexes in six-week old specific pathogen free chicken for all isolates were more than 0.7 and 1.2. Conlusions It is the first report of pathogenicity assessment of H5N1 virus circulating in Pakistan and all isolates were categorized to HPAI viruses through primary methods of pathogenicity test.

Future Challenges Related to Animal Biotechnology

Consumers and society may be least willing to accept the modern practices for genetic modification of animals. Public and animal welfare organizations criticism on animal cloning and transgenic modification is a hurdle in production of modified animal food. Ethical and moral concerns are also reduced the acceptability of such technology for the production of cheap and better animal food. Moreover, food industries are also very sensitive about the acceptance of consumer’s towards the products from genetically modified animals. Consumer is concern about the safety of such a food as compare to naturally produce animal food. Segregation and compulsory labeling about GMO’s food products by regulatory authorities also produce assumption of less safe and inferior food as compare to convention animal food products.

The Role of Biotechnology in Improvement of Livestock

This book examines how biotechnology can improve livestock breeding and farming, and thereby also animal products. In the first chapters the reader will discover which techniques and approaches are currently used to improve animal breeding, animal health and the value of animal products. Particular attention is given to reproduction techniques, animal nutrition and livestock vaccines that not only enhance animal health but also have a significant effect on human health by ensuring safe food procurement and preventing zoonotic diseases. In addition, modern biotechnology can increase not only productivity but also the consistency and quality of animal food, fiber and medical products. In the second part of the book, issues such as how animal biotechnology could affect the environment and the important topic of animal waste management are explored. In the concluding chapter, the authors discuss future challenges related to animal biotechnology. This work will appeal to a wide readership, from scientists and professionals working in animal production, to those in farm animal management and veterinary science

Modification of Animal Products for Fat and Other Characteristics

This chapter includes information about modification of animal products using biotechnology and the importance of different modifications on the natural composition. The species considered for modified products include beef and dairy cattle, sheep, goats, poultry, and a wide variety of fishes. Moreover, the discussion includes the importance of animal food, nongenetically engineered animal modified food products, genetically engineered animal modified food items primarily for meat, milk, or egg and genetically engineered animal food along the transgenic approach for animal welfare. Modern biotechnology can improve productivity, consistency, and quality of alter animal food, fiber, and medical products. The transgenic technology is potentially valuable to alter characters of economic importance in a rapid and precise way. The food safety issue related to genetic engineering is also included in this chapter. The harm of such modified food and transgenic strategy should also be understood by the reader along with its advantages. In this context, transgenic approaches in animal biotechnology are under discussion that ranges from animal food production to their adverse effects.