Search or add a thesis

Advanced Search (Beta)
Home > Molecular Characterization of Opportunistic Pathogens from Avian Mycoplasma Cases in Punjab

Molecular Characterization of Opportunistic Pathogens from Avian Mycoplasma Cases in Punjab

Thesis Info

Access Option

External Link

Author

Siddique, Abu Baker

Program

PhD

Institute

University of Agriculture

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2012

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/2106

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726688260

Asian Research Index Whatsapp Chanel
Asian Research Index Whatsapp Chanel

Join our Whatsapp Channel to get regular updates.

Similar


Respiratory tract infections are of great importance in poultry industry, causing heavy economic losses. Mycoplasma gallisepticum and Mycoplasma synoviae are the most pathogenic organisms of the respiratory tract. Other respiratory tract infections includes both viral pathogens (Newcastle disease virus, Infectious bronchitis virus, avian influenza virus) and bacterial pathogens (Salmonella pullorum, Escherichia coli, Avibacterium paragallinarum, etc) cause disease independently and in association with each other. The study was designed to check the possible role of Mycoplasma infections in disseminating other respiratory pathogens. Further, the different diagnostic techniques including serum plate agglutination (SPA) test, cultural isolation and polymerase chain reaction (PCR) were applied are compared for their capabilities for the identification of the pathogens. Serum Plate Agglutination (SPA) test was used for serological screening test for Mycoplasma species. Samples including oral/ nasal swabs, lungs trachea and air sac swabs were collected from sero-positive and sero-negative flocks. Cultural isolation was on Frey’s Modified medium for Mycoplasma isolation, embryonated eggs for viral isolation and blood agar for other bacterial isolation. Polymerase chain reaction and Reverse transcriptase-polymerase chain reaction was optimized for the molecular identification of bacterial and viral pathogens, respectively. Multiplex PCR was also optimized for the simultaneous detection of respiratory tract pathogens of both bacterial and viral pathogens including Mycoplasma gallisepticum, Mycoplasma synoviae, Newcastle disease virus, Avian influenza virus and Infectious bronchitis virus using specific primers. To resolve further variation among opportunistic pathogenic species, the PCR products were sequenced and phylogenetic analysis was carried out. In the present study 34 flocks showing respiratory distress were visited for serological screening of Mycoplasma involvement in respiratory distress cases. Out of 34 flocks visited 27 (79.1%) were serologically positive. Based on PCR based diagnosis, irrespective of serological status the highest involvement of bacterial pathogens recorded was MG (31.8%), followed by E. coli (20.7%), MS (7.9%) and Av. paragallinarum (5.3%). Moreover, in case viral pathogens recovery from respiratory distress cases was recorded maximum in NDV (24.9%) then IBV (4.3%) and AIV (1.5%). The multiplex PCR was efficiently optimized for the simultaneous detection of respiratory tract infections including Mycoplasma gallisepticum, Mycoplasma synoviae, Newcastle disease virus, Avian influenza virus and Infectious bronchitis virus. Mycoplasma gallisepticum amplified 720bp PCR product, while Mycoplasma synoviae, yielded 270bp product. In case of viral pathogens Newcastle disease virus was identified by amplifying 320bp product, Avian influenza virus, 1050bp PCR product and Infectious bronchitis virus yielded 1720bp band. DNA sequences of Mycoplasma gallisepticum, Mycoplasma synoviae and Newcastle disease virus was submitted to GenBank as Mycoplasma gallisepticum (lp- gene) strain ABSuafMG2011 partial sequence, (GenBank accession no. JN114112). For Mycoplasma synoviae (16SrRNA gene) strain ABSfsdMS2011 partial sequence, (GenBank accession no. JN638722). While for Newcastle disease virus (Fusion gene) stains ABSuafND2011 partial sequence, (GenBank accession no. JN160608) and strain ABSfsdND2011 partial sequence (GenBank accession no. JN377950) In conclusion, the incidence of respiratory tract pathogens in sero-positive flocks for Mycoplasma was found higher as compared to sero-negative flocks. The true prevalence of the Mycoplasma infections is reflected by combining PCR results with SPA test. The present study also documented the involvement of indigenous strains of MG, MS and NDV in the respiratory distress cases. Multiplex PCR was successfully optimized for the simultaneous and early detection of respiratory tract infections.
Loading...
Loading...

Similar Books

Loading...

Similar Chapters

Loading...

Similar News

Loading...

Similar Articles

Loading...

Similar Article Headings

Loading...

نامہ بروں سے ربط نہ بادِ صبا سے ہے


نامہ بروں سے ربط نہ بادِصبا سے ہے
بس آہ بھر کا فاصلہ غم آشناؐ سے ہے

پوچھا کہ کس کے دم سے ہے خوشبو چمن چمن
بولی صبا حضورؐ کی زلفِ دوتا سے ہے

ہر شب کا حسن آپؐ کی والیل زُلف سے
ہر دن کی آب و تاب رُخِ والضحیٰ سے ہے

بدرالدجیٰؐ کے نور سے مہتاب کی چمک
تاروں کی روشنی بھی اُسی کی ضیا سے ہے

اُس ذاتؐ کے طفیل ہے دارین کا نظام
دل کی بھی کائنات اُسی دلرُباؐ سے ہے

کس کے لیے سجائی گئی بزمِ کائنات
دلداریِ خدا یہ حبیبِ خداؐ سے ہے

فرشِ زمیں سے عرشِ بریں کی حدود تک
’’سارے جہاں کا حُسن دمِ مصطفی سے ہے‘‘

قرآں کی آیتوں میں کیا ان کا تذکرہ
خالق کو پیار آپؐ کی اِک اِک ادا سے ہے

روزِ ازل سے اِسمِ محمدﷺ خدا کے ساتھ
یہ اِنتہا کو پہنچا ہوا ابتدا سے ہے

عرفانؔ جو نبیؐ کا ہوا وہ خدا کا ہے
الفت رسولِ پاکؐ کی الفت خدا سے ہے

Menstrual Cycle in Female Workers

Menstrual disorders are abnormalities that occur in the menstrual cycle. There are various menstrual disorders that women can experience, ranging from too little or too much menstrual blood, painful menstruation, to depression before menstruation or premenstrual dysphoric disorder. Physical activity is divided into three levels, namely light, moderate and heavy physical activity. Physical activity of heavy intensity can cause physiological disorders of the menstrual cycle. Presence of menstruation (amenorrhoea), thinning of the bones (osteoporosis), menstrual irregularities or intermenstrual bleeding, abnormal growth of the uterine wall, and infertility.

Paramecium Diversity and Phylogeny: Three Loci Molecular Characterization

Ciliated protozoan are an important bioindicators for pollution and have long been used for environmental biomonitoring, particularly in water purification plants and in activated sludge processes. Each species of ciliates has its own physical and chemical valences and therefore based on the indicator values of representative ciliates, degree of pollution of a particular water body can be determined. Among ciliates Paramecium has become a privileged model for the study of “species problem” particularly in the case of “Paramecium aurelia complex” that has been intensely investigated. Despite extensive studies taxonomy of Paramecium is still changing. The major problem is uneven sampling of Paramecium with relatively few representatives of each species. Continued discovery of new species from various origins (worldwide) are proof of the fact that the list of Paramecium species is not yet complete. Hence, the addition of new species by more sampling is needed to resolve the phylogeny within the Paramecium genus. For this purpose, present study was aimed at molecular characterization of Paramecium species from water samples originating from various regions of Punjab province of Pakistan. Fragments of 18S rDNA, ITS1-5.8S-ITS2-5’LSU rDNA, COII, Hsp70 and Histone H4 genes were used as molecular markers for the phylogenetic analysis of ten locally isolated strains of Paramecium species including a standing-alone FT8 strain previously isolated by Shakoori et al. (2014). The nucleotide sequences of PCR products of different molecular markers of various isolates (FT2.1, FT3.1, FT4.1, FT5.1, FT6.1, FT7.1, FT9.1, FT10.1 and FT11.1) were compared with the available sequences of these markers in other Paramecium species from GenBank. Phylogenetic trees based on all molecular markers showed that all nine strains (FT2.1, FT3.1, FT4.1, FT5.1, FT6.1, FT7.1, FT9.1 FT10.1 and FT11.1) had very close relationship with P. primaurelia except for FT8 strain. FT8 showed its unique position in comparison to all other species in the phylogenetic trees, so became the main focus of our study. Phylogeny of this strain was further carried out with PiggyMac sequence, where it again behaved differently as compared to other species. Sexual behavior, immaturity and maturity periods of FT8 were analyzed by performing daily reisolations that revealed three unique characteristics of this strain; 1) autogamy as only source of the exchange of genetic material 2) no clumps formation before conjugation that is the prerequisite of sexual process, 3) selfing among reactive cells. Second and third characteristics of this species turned our attention towards its mating types (“odd” 0 and “even” E). Why agglutination does not occur in this strain like all other Paramecium species? Or maybe there are circadian rhythms of mating xv types? In order to resolve this mystery, an experiment following circadian rhythms of completely light and completely dark cycles was performed multiple times during a period of one month. However, every time no conjugation upon mixing but selfing in the original cultures was observed. Results of this study could not prove the existence of mating types in FT8 strain but off course detailed analyses at genetic level is required for precise knowledge. Another important aim was to count the number of micronuclei of this unique species with and without Spt5-GFP signals. Four number of MICs on an average were observed, whereas two, three and five MICs were also seen in some karyonides. However, GFP signal was found to be activated till the skein formation of conjugating pairs. No signal in micronuclei of later stages could be observed. Finally, number of events of whole genome duplications (WGDs) of FT8 strain were studied by counting the number of paralogs for highly expressed, highly conserved L8 and S13 ribosomal protein genes. Total three number of genes (L8FT8-1, L8FT8-3 and L8-FT8-4) with L8 amplifications and two genes (S13FT8-1 and S13FT8-2) with S13 after extensive screenings were found. Number of genes obtained from amplifications results supported that this species has not gone through any additional WGD event. However, whole genome sequencing of this species would resolve the complete and precise information of its number of WGDs. Finally, based on the phylogenetic trees of all molecular markers including L8 and S13 genes, it can be concluded that despite occupying unique position in the phylogenetic tree, FT8 was closer to P. multimicronucleatum than to P. caudatum.