About
55
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Introduction
My research has been focused on understanding the molecular basis of thermal instability and/or attenuation of temperature-sensitive Mycoplasma synoviae vaccine strain MS-H (Vaxsafe MS, Bioproperties, Australia). Research work included proteomic and genomic analyses to underpin attenuating factors, development of genetic manipulation tools (e.g., oriC based shuttle vectors) to study the role of putative genetic factors in attenuation of M. synoviae, and development of recombinant MS-H vaccine(s) expressing immunodominant genes of economically important diseases-causing-pathogens of poultry.
Current institution
Additional affiliations
December 2014 - present
Position
- Professor (Assistant)
Description
- Teaching under- and postgraduate courses of Microbiology.
June 2009 - August 2014
June 2011 - August 2014
Position
- Casual employee
Description
- Assisted Veterinary Science, undergrad students in practical classes of Avian diseases
Education
June 2009 - July 2014
University of Melbourne, Australia
Field of study
- Veterinary Microbiology
September 2004 - September 2006
University of Agriculture Faisalabad, Pakistan
Field of study
- Veterinary Microbiology
December 2000 - September 2004
University of Agriculture Faisalabad, Pakistan
Field of study
- Veterinary Medicine
Publications
Publications (55)
The growth of fungus in grains and feed is favored by improper humidity and temperature during storage contributing to loss of grain quality, infections among animals and humans and production of mycotoxins. Therefore, the current study was aimed to isolate and identify fungal species among stored grains and feedstuff of Potohar region of Pakistan....
Simple Summary
The harmful effects of microplastic (MP) exposure on aquatic animals have been extensively studied; however, there is a lack of research on its impact on poultry. To address this gap, the present study aimed to evaluate the effects of MP exposure on the growth performance and gut microbiota of chickens. The findings of the study reve...
Feral birds residing close to urban settings exhibit higher immunocompetence against environmental pathogens than their counterparts in rural areas. In this study, we comprehensively evaluated the immunocompetence of five specific feral bird species and investigated the potential for interspecies transmission and pathogenicity of Avian orthoavulavi...
The detection of toxic molecules has gained imperial attention to protect living organisms and their environment. The broadly explored sensor behaviour of triazine-based C3N4 reveals their ultra-high sensitivity towards numerous toxic molecules e.g., chemical welfare agents. However, another potential analogue of C3N4 composed of heptazine exhibits...
Swertia bimaculata (SB) is a medicinal herb in China having an array of therapeutic and biological properties. This study aimed to explore the attenuating effect of SB on carbon tetrachloride (CCl 4) induced hepato-toxicity by regulation of gut microbiome in ICR mice. For this purpose, CCl 4 was injected intraperitoneally in different mice groups (...
The yaks that inhabit the Tibetan plateau are a rare breed that is closely related to local economic development and human civilization. This ancient breed may have evolved a unique gut microbiota due to the hypoxic high-altitude environment. The gut microbiota is susceptible to external factors, but research regarding the effects of different feed...
Campylobacteriosis is caused by Gram-negative and spiral-shaped microaerophilic Campylobacter bacteria. Different avian hosts are commonly infected with Campylobacter species. Among 16 Campylobacter species, infections are mostly caused by thermophilic Campylobacter jejuni and Campylobacter coli. C. jejuni and C. coli are well adapted to the avian...
Campylobacteriosis is caused by members of the Campylobacteriaceae family which are Gram-negative and spiral-shaped microaerophilic bacteria. Different avian hosts, including chickens, turkeys, ducks, and geese, are commonly infected with Campylobacter species. Among 16 Campylobacter species, infections are mostly caused by thermophilic Campylobact...
Methicillin-resistant Staphylococcus aureus (MRSA) is a clinically prevalent bacterium and is resistant to many drugs. Genetic factors such as mec genes are considered to be responsible for this resistance. Recently, Staphylococcal Cassette Chromosome mec (SCCmec) element mutations produced mecC, a new genetic variant that encodes a transpeptidase...
Pesticides are widely used to control crop diseases, which have made an important contribution to the increase of global crop production. However, a considerable part of pesticides may remain in plants, posing a huge threat to animal safety. Thiram is a common pesticide and has been proven that its residues in the feed can affect the growth perform...
Cutaneous candidiasis is one of the most prevalent mycotic infections caused by Candida species. The severity of infection mounts faster when the species shows antifungal resistance. In the current retrospective study, we aimed to analyze the occurrence, causes of cutaneous candidiasis, and antifungal susceptibility pattern of Candida isolates from...
Foot-and-mouth disease virus (FMDV) is one of highly contagious pathogens of cloven-footed animals, attaches with host cell surface receptors, including the integrins and heparan sulfate, for entry into the epithelial cells. FMDV is endemic in Pakistan, but limited studies are available on the interaction of local FMDV serotypes with host-entry fac...
Background
Avian influenza H9 is endemic in commercial and backyard poultry in Pakistan and is a serious occupational health hazard to industry workers. This study aimed to determine the seroprevalence of avian influenza H9 infection in people working with poultry in Rawalpindi, Pakistan and assess the measures they took to protect themselves from...
The Mycoplasma synoviae (MS) vaccine strain MS-H harbours a frameshift mutation in oppF1 (oligopeptide permease transporter) which results in expression of a truncated OppF1. The effect of this mutation on growth and attenuation of the MS-H is unknown. In this study, the impact of the mutation on the vaccine phenotype was investigated in vitro by i...
Mycoplasma synoviae is an economically important avian pathogen worldwide, causing respiratory disease, infectious synovitis, airsacculitis and eggshell apex abnormalities in commercial chickens. Despite the widespread use of MS-H as a live attenuated vaccine over the past two decades, the precise molecular basis for loss of virulence in this vacci...
Pathology of hydropericardium syndrome (HPS) induced by fowl adenovirus-4 (FAdV-4) in ostriches was compared with that of broiler chicken kept as standard. A 10 6.25 /ml/bird (chicken lethal dose 50 (CLD 50)) was injected to ostriches and broiler (control) and were clinically observed for 7 days. Our results showed that a peak cloacal temperature w...
Background:
Avian influenza H9 is endemic in commercial and backyard poultry in Pakistan and is a serious occupational health hazard to industry workers. This study aimed to determine the seroprevalence of avian influenza H9 infection in people working with poultry in Rawalpindi, Pakistan and assess the measures they took to protect themselves fro...
Fowl adenovirus has been reported in many countries and is a contagious agent related with inclusion
body hepatitis (IBH) and hydropericardium syndrome (HPS) in chickens worldwide. Identification of
fowl adenovirus (FAdV) serotype is of most importance in epidemiological studies of disease outbreak
and the adaptation of vaccine strategies. In spite...
Fowl adenovirus has been reported in many countries and is a contagious agent related with inclusion
body hepatitis (IBH) and hydropericardium syndrome (HPS) in chickens worldwide. Identification of
fowl adenovirus (FAdV) serotype is of most importance in epidemiological studies of disease outbreak
and the adaptation of vaccine strategies. In sp...
To cite this paper: Rahman, S.U., S. Ahmad, H. Khan, M.A. Shahid and I. Ahmad, 2018. Expression and analysis of two-partner secretion system of Neisseria meningitidis using heterologous host Escherichia coli. Int. Abstract In Two-Partner Secretion (TPS) systems, the outer membrane-embedded TpsB transporter is dedicated for the transport of a large...
The temperature-sensitive (ts⁺) Mycoplasma synoviae vaccine strain MS-H harbors a non-synonymous mutation which results in Glycine to Arginine substitution at position 123 in the highly conserved glycine-rich motif of Obg-fold in the GTP-binding protein Obg. In-silico analysis of the wild-type and mutant Obgs of M. synoviae has indicated that this...
Analysis of Obg expression in E. coli and M. synoviae using SDS-PAGE and immunoblotting.
(A) SDS-PAGE of E. coli lysates from noninduced culture, induced culture, supernatant from induced culture, and affinity purified Obg-MBP. Arrows indicate location of the recombinant M. synoviae Obg and MBP proteins in E. coli, respectively. (B) Immunostaining...
Expression of M. synoviae Obg in E. coli and production of Obg-MBP antiserum in chickens.
(DOCX)
Northern analysis of total RNA (~ 25 μg/lane) from M. synoviae strain MS-H, MS-H-C28, MS-H-T75, MS-H-T78 and MS-H-T90.
Northern blots were hybridised with DIG-labeled vlhA-obg probe (spanning the joining site of vlhA promoter with obg CDS), obg specific probe and vlhA coding sequence specific probe. Arrowheads indicate the location of the specific...
Comparison of Obg expression in MS-H and 86079/7NS.
Immunostaining at two different concentrations of whole cell proteins (i.e. 1/3 and 2/3 of that loaded onto SDS-PAGE as shown in Fig 4E) of each strain/transformant probed with polyclonal rabbit serum against M. synoviae Obg_1 peptide. MW, protein marker (Precision Plus Protein, Dual Color, Bio-Ra...
Oligonucleotides used in this study for PCR amplifications and probe synthesis.
(DOCX)
Splicing by overlap extension PCR.
(DOCX)
Background:
The bacterial pathogen Mycoplasma synoviae can cause subclinical respiratory disease, synovitis, airsacculitis and reproductive tract disease in poultry and is a major cause of economic loss worldwide. The M. synoviae strain MS-H was developed by chemical mutagenesis of an Australian isolate and has been used as a live attenuated vacci...
Mycoplasma synoviae, an important poultry pathogen, belonging to the class Mollicutes, causes airsacculitis, synovitis, decreased egg production and produces significant economic losses. Efforts to determine M. synoviae virulence factors and their role in pathogenicity require suitable tools for genetic manipulation of this pathogen. This study des...
Temperature-sensitive (ts+) vaccine strain MS-H is the only live attenuated M. synoviae vaccine commercially available for use in poultry. With increasing use of this vaccine to control M. synoviae infections, differentiation of MS-H from field M. synoviae strains and from rarely occurring non-temperature-sensitive (ts-) MS-H revertants has become...
Mycoplasma synoviae strain MS-H, developed by chemical mutagenesis of the Australian field strain 86079/7NS, is a live temperature-sensitive (ts (+)) vaccine used for control of M. synoviae infection in poultry worldwide. Genetic basis of temperature sensitivity and attenuation of MS-H has not been revealed thus far. Comparison of the complete geno...
Mycoplasma synoviae infections result in significant economic losses in the chicken and turkey industries. A commercially available live temperature-sensitive (ts (+)) vaccine strain MS-H has been found to be effective in controlling M. synoviae infections in commercial layer and broiler breeder farms in various countries, including Australia. Dete...
Mycoplasma synoviae infections result in significant economic losses in the chicken and turkey industries. A commercially available live temperature-sensitive (ts+ ) vaccine strain MS-H has been found to be effective in controlling M. synoviae infections in commercial layer and broiler breeder farms in various countries including Australia. Detecti...
The present study was contemplated to find out the epidemiology of mastitis in lactating cattle and buffaloes in tehsil Burewala, Pakistan. For this purpose, a total of 673 animals (n=291 cattle, n=382 buffaloes) from 300 livestock farmers were tested using Surf Filed Mastitis Test (SFMT) for the presence of mastitis. A higher prevalence (24.60%) o...
Antibiotic residues in meat pose various human health hazards to susceptible individuals including gastrointestinal disturbances, teratogenic risks, allergic reactions and development of resistant pathogens for human and animals. Therefore monitoring the level of antibiotic residues is of utmost importance to safeguard human health. Various methods...
Present study was performed to determine the effects of physical and chemical agents on infective potential of highly pathogenic avian influenza (HPAI) H5N1 (local strain) virus recently isolated in Pakistan during 2006 outbreak. H5N1 virus having titer 10(8.3) ELD(50)/ml was mixed with sterilized peptone water to get final dilution of 4HA units an...
The present study was contemplated to find out the epidemiology of mastitis in lactating cattle and buffaloes in tehsil Burewala, Pakistan. For this purpose, a total of 673 animals (n=291 cattle, n=382 buffaloes) from 300 livestock farmers were tested using Surf Filed Mastitis Test (SFMT) for the presence of mastitis. A higher prevalence (24.60%) o...
Abstract
Present study was performed to determine the effects of physical and chemical agents on infective potential of highly pathogenic avian influenza (HPAI) H5N1 (local strain) virus recently isolated in Pakistan during 2006 outbreak. H5N1 virus having titer 10<sup>8.3 </sup>ELD<sub>50</sub>/ml was mixed with sterilized peptone water to get fi...
A respiratory disease outbreak was reported in layer flocks in and around Sammundri area of Punjab. Postmortem of the affected birds were performed and morbid organs (trachea, lungs, air sacs, spleen, proventriculus and brain) were collected and processed for isolation and identification of etiological agent. The samples were inoculated in 9-days o...
Non-vaccinated commercial layer farms against any subtype of avian influenza virus were visited, having respiratory and other problems confusing with avian influenza to collect tissue samples and swabs for isolation of the virus. Samples were processed and inoculated into embryonated chicken eggs. Harvested materials were subjected to haemagglutina...
The study was conducted on lactating cattle and buffaloes in tehsil Burewala, district Vehari, Pakistan. The purpose of study was to isolate different types of microorganisms associated with mastitis and to see the effectiveness of different antibiotics against these isolates. Relative prevalence of Staphylococcus aureus was the highest (50% and 53...
Mastitis continues to be a major economic issue for dairy producers all over the world. Staphylococcus aureus is frequently isolated from bubaline mastitis and brings about glandular tissues changes obliterating the milk producing cells. Elimination of this organism from dairy herds requires treatment of infected mammary glands with antimicrobial a...
The present study was conducted to monitor the presence of antibiotic residues in poultry meat. Swab Test on Animal Food (STAF) employing Bacillus subtilis as test organism on nutrient agar with 0.4% dextrose at pH 7, was used. A spore suspension of 2x107 spores mL-1 was used in 100 mL of nutrient agar to make STAF plates. A total of 100 tissue sam...
Nasopharyngeal swabs from healthy cattle (n = 100) and buffalo (n = 100) were collected and inoculated on the selective media for Pasteurella multocida i.e., Casein Sucrose Yeast agar supplemented with blood and incubated at 37°C for 48 h. Isolated organisms were identified on the basis of cultural, morphological and biochemical characteristics. Th...
A respiratory disease outbreak was reported in layer flocks in and around Sammundri area of Punjab. Postmortem of the affected birds were performed and morbid organs (trachea, lungs, air sacs, spleen, proventriculus and brain) were collected and processed for isolation and identification of etiological agent. The samples were inoculated in 9-days o...
Questions
Questions (10)
We are working on a postbiotic preparation to see its effect on rumen health. We need a few aerobic or facultative bacteria which can be used in viable count. Even if we use a selective agar, the target bacterium can't be excluded from other few organisms which will also grown on the agar plate. So what should be target bacterium and the modification of the viable count protocol? We don't have anaerobic bacterial culture facility, so have to rely on some aerobic or facultative anaerobe which we can grown in our lab in routine aerobic incubators.
How can we determine the concentration of antibiotic pellets available with the antibiotic cartridge? For example, Meropenem concentration from Oxoid is 10 ug per disc, but what would be the concentration of the pellet included in the cartridge.
Normally dN/dS ratios are calculated and interpreted as below one negative selection, above 1 is positive selection and 1 means neutral selection. How to interpret dS/dN ratios? Programs like SNAP provides dS/dN graphs and ratios.
For example:
Averages of all pairwise comparisons: ds = 0.1678, dn = 0.4090, ds/dn = 0.4072, ps/pn = 0.4707
Please see image as well.
Can somebody explains it in simple words as I am not much familiar with this?
To perform HRM analysis for genotyping on CFX96 machine for the first time, BioRad recommends to use HRM calibration kit along with Precision melt analysis software. I am just wondering whether simply using any good quality DNA binding dye along with standard PCR reagents will work or not?
I have been using Corbett Rotorgene 6000 for HRM analysis. In my new lab, we have BioRad CFX96 machine but its software (Precision melt analysis) is very expensive (5000 USD) unlike rotorgene which uses the same software for HRM analysis as used for other Real time data analysis. Is there any alternative of buying 5000 dollar software for CFX96 for performing HRM analysis?
Also, we do have Applied Biosystems® 7500 Real-Time PCR Systems. I don't know if this can be used for HRM analysis with similar/useful sensitivities as Rotorgene machine. Any help in this regard would be highly appreciated.
Please also comment if ABI 7500 real time system also requires a separate software for HRM analysis (like CFX96 system) or it can be done with the software used for routine real time data analysis software.
For example: if one strain has starting inoculum (CCU/ml) of log10 3.5, second with 4 and third with 3, is it still possible to measure and quantitate the difference in their growth rate? If yes then how? One way is to measure doubling time for all strains at some specific time interval. But main question is whether slight variation, as mentioned above, has any importance on the measured doubling time? Sometimes it is very difficult to standardize all cultures at exactly the same concentration especially mycoplasma.
We normally use 4X protein sample lysis buffer containing 250 mM Tris (pH 6.8), 40% glycerol, 10% β-mercaptoethanol, 9.2% SDS, 0.02% bromophenol blue. I have used 2-D quant kit (GE, Health care) for protein quantification for 2D samples (which normally doesn't contain glycerol, bromophenol etc).
I was just wondering if someone has used this kit with SDS-PAGE sample preparation buffer (as described above) which contains glycerol and bromophenol blue (two things which 2-D kit doesn't claim any tolerance or compatibility with, otherwise excellent in presence of SDS, CHAPS or other constituents of buffers).
I want to use this 2-D kit for quantification of my protein samples which are normally prepared in sample lysis buffer as described above but has never used this kit for these samples. Any suggestion or experience regarding this would be highly appreciated.
After adjusting concentration of whole cell proteins from different bacterial strains, I want to measure relative expression of a particular protein by Western blot (DAB, Diaminobenzidine method). First I stain SDS-PAGE gels with coomassie and then take image with Gel Logic 1500 Imaging System (Kodak), choosing 16 or 8 bit TIFF image. I am using Image J to measure band densities, for few selected bands, to measure and normalize amount of whole cell proteins to be loaded into the wells. As such there is no known protein to serve as an internal control for this bacteria, therefore I randomly select few protein bands. Is it an acceptable method by peer reviewed journals?
