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Home > Transcriptome Profiling of the Virus-Induced Innate Immune Responses in Chickens Challenged With Newcastle Disease Viruses of Different Pathogenicity

Transcriptome Profiling of the Virus-Induced Innate Immune Responses in Chickens Challenged With Newcastle Disease Viruses of Different Pathogenicity

Thesis Info

Access Option

External Link

Author

Aziz-Ul-Rahman

Program

PhD

Institute

University of Veterinary and Animal Sciences

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Microbiology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/11506/1/Aziz-ul-Rahman%20%20Microbiology%202019%20uvas%20lhr%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727627838

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Newcastle disease is an acute highly contagious disease, responsible for serious economic losses to poultry industry worldwide particularly in disease endemic regions such as Pakistan. Subsequent to evolution, with the passage of time, novel genotypes and sub-genotypes are emerging worldwide raising concerns on efficacy of vaccine strains and, therefore ascertain candidate vaccines that can provide protection much better than those used before. Now a day, with the advancement in reverse genetics system, efforts are ongoing to construct a vaccine strain that, beside specific immunity, may have the potential to enhance innate immunity and therefore a better use against a viral infection. A much has been elucidated about clinical pathogenicity of NDVs of different pathotypes and genotypes in their susceptible hosts (e.g., chickens); nevertheless, corresponding to innate immune response, there is a paucity of data regarding comparative assessment of transcriptome profile in birds challenged with NDVs of varying pathogenicity to better unleash a comparative rate of expression of differentially expressed genes (DEGs) and their subsequent use in the development of novel vaccines through reverse genetics system. Therefore, the current study was conducted to 1) determine biologic, genomic and genotypic characterization of NDV strains isolated from waterfowl and pigeon independently, 2) a clinico-pathological assessment of these strains in chickens and pigeons and, 3) a comparative assessment of innate immune response-related genes that become either up-regulated or down-regulated upon exposure to NDV strains of varying pathogenicity in chicken using Illumia HiSeq-2500. Biological, genomic and genotypic analysis categorized the Anseriformes-originated NDV isolate as velogenic strain (sub-genotype VIIi) whereas pigeon-originated NDV isolate as mesogenic strain (sub-genotype VIm). Infectious potential of velogenic and mesogenic NDV strains was assessed in broiler chickens and pigeons, separately. For each of virus exposure, variations in the severity and duration of infection were observed among challenged and contact chickens and pigeons. Chicken were found more susceptible to velogenic strain while pigeons were found more susceptible to mesogenic strain. Comparative transcriptome analysis revealed expression profile of innate immune response-related genes in lungs and spleen tissues of chickens infected with velogenic and mesogenic strains. Following Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, a total of 109 DEGs were commonly expressed in tissue samples (lung-A, spleen-A, lung-P and spleen-P). Genes associated with metabolic and cellular processes such as innate immune system associated signalling pathways, RIG-I, Toll-like, NF-Kappa β and MAPK signalling pathways for activation of interferonstimulated genes (ISGs) were more abundant. DAVID-based annotation classified these 109 genes into five major categories that included interferon stimulatory genes (ISGs) (n = 37), lymphoid tissue innate immunity (n = 30), inflammatory responses associated genes (n = 29), genes for regulation of adaptive immune components (n = 12) and toll like receptor (n = 1). While comparing lung tissues (lung-A vs lung-P), analysis revealed a significant variation in expression of 101 of 109 DEGs, whereas, 106 DEGs had significant variations in expression for spleen tissues (spleen-A vs spleen-P). Out of 109 DEGs, for lung tissue, six genes (CD22, CR2, FOXS1, ITGA6, KMTSE, LYST, USP15 and USP33) had a non-significant difference in rate of expression while it was three genes (IL15, KLHL13, MKO7) for spleen tissue. Hence, post-infection with NDVs of varying pathogenicity, study highlights differential expression of innate immunity related genes where a candidate gene could be selected to construct vaccine strains through reverse genetics system with cumulative potential to enhance immunity and combat challenge from field viruses in the future.
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ام المومنین سیدہ ماریہ قبطیہؓ

ام المومنین سیدہ ماریہ قبطیہؓ
سیدہ ماریہ قبطیہ بنت شمعون کو بطور ہدیہ شاہ مقوقس نے آپؐ کی خدمت میں بھیجا تھا۔(تبیان القرآن جلد ۱۲ ص ۱۰۳) پر ہے’’ دو بہنیں ماریہ اور سیرین ‘ ایک ہزار مثقال سونا، بیس ملائم کپڑے، خچر(دلدل) ایک درازگوش گدھایفغور یا عفیر تھا اور اس کے ساتھ ایک خصی شخص جس کا نام ثابور تھا اور ایک بوڑھا شخص جو ماریہ کا بھائی تھا ۔اس نے مدینہ پہنچ کر اسلام قبول کر لیا تھا۔مقوقس مصر کے ہر قل (قیصر روم) کا نائب تھا۔ مقوقس اس کا لقب تھا۔مسلمان مورخین نے اس کا نام جریج بن سینا القبطی بتایا ہے۔ مقوقس نے تحائف بھیجے جن میں دو قبطی النسل کی معزز خواتین، ایک خلعت، سواری کے لیے گھوڑا( افرازہ نامی) ایک سفید خچر( دلدل) اور ایک گدھا یفغور یا عفیر شامل تھے۔(دائرۃ معارف الاسلام ج۲۱ص ۴۵۵) لیکن صاحب مدارج النبوہ ( مدارج النبوہ جلد دوم ص ۲۸۳) لکھتے ہیںکہ ان تحائف میں ایک نیزہ ،بیس قد کا لباس اور ہزار مثقال سونا، مزید لکھتے ہیں کہ حاطب بن بلتعہ کو سو مثقال سونا اور پانچ کپڑے بطور انعام کے دئیے‘‘۔ آپﷺ نے حضرت ماریہ کو حرم نبوی میں داخل فرمایا۔ان سے جناب ابراھیم پیدا ہوئے۔ساتویں روز ان کا عقیقہ کیا گیا۔سر منڈھا کر بالوں کے برابر چاندی تول کر صدقہ کی۔ دو مینڈھے ذبح کیے۔جناب ابراھیم ۱۷ یا ۱۸ ماہ کی عمر میں ہجرت کے دسویں سال انتقال کر گئے۔آپﷺ نے فرمایا: بخدا !! ابراھیم ہم تمھاری موت سے نہایت غمگین ہیں،آنکھ رو رہی ہے اور دل غم زدہ ہے مگر ہم ایسی کوئی بات زبان سے نہ کہیں گے جس سے ہمارا رب راضی نہ ہو‘‘ ۔
رسمِ باطل کا خاتمہ: جس روز آپ کا انتقال ہو ا، اتفاق سے سورج کو گہن لگا۔عرب سمجھتے تھے کہ سورج گرہن...

حضور انورﷺ کی رحمت کی امتیازی خصوصیات

The many aspects of the Holy Prophet’s (PBUH) personality has been defined by Allah himself in Quran many times, keeping in view the all angles of the Holy Prophet (PBUH) Character blessings for all worlds and times. The Rehmatul-il-Almeen is infinite. The Holy Prophet (PBUH) has proved such a huge chain by his character. The blessing of the Holy Prophet (PBUH) is such a supreme that, it could not be defined in such an article, but here it has been tried to throw light on some features of Holy Prophet (PBUH) blessing. As Quran says that the secret of man on the path of the Holy Prophet (PBUH). Today same way, man regardless of color, race and religious should know that the wounds of humanity could be headed by following the route of the Holy Prophet (PBUH) Sunnah. All the other ways will lead to cul-de-sac.

Effect of Bacillus Subtilis and Sodium Butyrate on the Morphometry of the Small Intestine and Immune System in Healthy and Salmonella-Challenged Broiler Chickens

Supplementation of Bacillus subtilis and microencapsulated sodium butyrate in the feed is being practiced as a substitute for antibiotics growth promoters. An expansive range of encouraging health-related properties exhibited by B. subtilis and SB has been published, but their exact effect on gut and immune system is not completely understood. Consequently, the evaluation of B. subtilis and SB as feed supplements is desired. To achieve this goal, the present study was aimed to investigate the effects of B. subtilis and SB on performance, immune system, gut and lymphoid organs microarchitecture in healthy and Salmonella-challenged broiler chickens. In the first experiment the research was targeted to investigate the effects of B. subtilis on performance, immune system, gut and lymphoid organ microarchitecture in broilers. A total of 120 d-old broiler chicks were randomly distributed into four groups, each group with three replicates containing 10 birds per replicate. The birds were fed a corn-soy-based basal diet (BD, control) or BD supplemented with 10% zinc bacitracin (ZnB), and 0.05g/kg or 0.1g/kg of B. subtilis, respectively. On d 21 and 35, six birds from each group were killed to collect blood and visceral organs (thymus, spleen, bursa of Fabricius, liver and small intestine). Parameters evaluated included growth performance, immune responses, relative organ weights, lymphoid organs and gut mucosal morphometry, intraepithelial lymphocytes (IEL) count and goblet cell histochemistry in mucosa. Results showed that the group fed 0.1g/kg of B. subtilis had superior (P < 0.05) mean body weight and weight gain, and lower FCR compared to the nonsupplemented or ZnB-fed groups. The BS-0.1 group revealed higher antibody titer against Newcastle disease (ND) virus and the supplemented groups against sheep RBCs (SRBCs) on d 35. Cell-mediated immune response post-phytohemagglutinin-P injection was attained (P < 0.05) by birds in the BS-0.1 group at 24 h, and by both the BS-0.1 and BS-0.05 groups at 48 and 72 h compared to the ZnB and control groups. The BS-0.1 group gained higher (P < 0.05) relative bursal weight on d 21 compared to the other groups. Compared to the control group, the liver, spleen and thymus weighed more (P < 0.05) in the experimental groups on d 35. The histomorphological study revealed increased (P < 0.05) thymus cortical width, and cortex/medulla ratio in the BS-0.1 group compared to the control. The area of the bursal follicles and germinal centers of the spleen also improved (P < 0.05) in the BS-0.1 group compared to the control. Compared to the ZnB and control, higher (P < 0.05) villus height, villus surface area and villus crypt ratio of the duodenum and jejunum were recorded on d 21, and higher (P < 0.05) villus height of the duodenum and ileum was noted on d 35 in the BS-0.1 and BS-0.05 groups. The number of goblet cells having acid mucin was significantly higher in the ileal mucosae of the BS-0.1 group chickens compared to the ZnB and control. In conclusion, B. subtilis type probiotics effectuated better growth performance, improved immune system and modulated morphology of lymphoid organs and gut mucosa in broilers. The second experiment was carried out to evaluate the effects of sodium butyrate on growth performance, immune status, organ weights and the microarchitecture of lymphoid organs and the small intestine compared to the effects brought about by an antibiotic. The cell-mediated immune response at 48 h post-phytohemagglutinin-P injection, and antibody titer against NDV and sheep RBCs on d 35 was higher (P < 0.05) in SB-1 chicks compared to those in the ZnB and control groups. Higher (P < 0.05) weight gain, and lower (P < 0.05) FCR were attained by the supplemented groups compared to the control. The thymus and spleen weighed more (P < 0.05) in the SB-1 group and bursa registered more (P < 0.05) weight in both SB groups compared to the control. On d 21, areas of the thymus medulla and the spleen germinal centers were larger (P < 0.05) in SB-1 chicks compared to ZnB and control chicks. The VH and VSA increased (P < 0.05) in the duodenum and jejunum in both SB groups on d 21, and in SB-1 on d 35 compared to the ZnB and control groups. The villus to crypt ratio was higher (P < 0.05) in the duodenum in SB-1 chicks compared to ZnB and control chicks. On d 35, VH in all segments and VSA in the duodenum and jejunum increased (P < 0.05) in SB-1 chicks compared to ZnB and control chicks. Statistically, IEL count was not significant among supplemented groups. On d 21, the number of goblet cells containing acidic mucin increased (P < 0.05) in all the segments of the small intestines in the SB-1 group compared to the control group and on d 35 in the ileum compared to the other groups. In conclusion sodium butyrate elicited better growth performance, improved immune system and modulated the morphology of lymphoid organs and the gut mucosa in broiler chickens. The third experiment was focused to assess the effect of B. subtilis and SB on gut development, growth performance and immune system in broilers challenged with S. Gallinarum. Better growth performance was reported in the supplemented groups compared to the NC-S group due to better feed efficiency. The B. subtilis-supplemented group exhibited higher (P < 0.05) cellular immunity and antibody titer against NDV compared to the PC-S and NC-S groups. Furthermore, B. subtilis- and SB-supplemented groups reflected higher (P < 0.05) relative thymus and bursa weights, and improved microarchitecture of the lymphoid organs compared to the NC-S group. On d 21, villus surface area in the jejunum and ileum increased (P < 0.05) in sodium butyratetreated birds. The crypt depth of the jejunum decreased (P < 0.05) in B. subtilis and sodium butyrate groups compared to NC-S and PC-S groups. On d 35, the villus height, villus surface area and VH:CD ratio of the duodenum increased (P < 0.05) in the supplemented groups compared to the NC-S group. The FCR, Salmonella population in ceca and mortality were higher (P < 0.05) in the NC-S group. In conclusion, the prophylactic use of the B. subtilis probiotic and SB alleviated stress associated with Salmonella Gallinarum infection and improved performance, immune function, lymphoid organs and gut mucosal development in infected broilers. Further analyses are needed to reveal the mechanism(s) by which B. subtilis and sodium butyrate produce such effects.