Papers by Dr mubashir Hussain
Eastern Mediterranean Health Journal, Sep 1, 2007
Research Square (Research Square), Oct 26, 2022
The detailed description of parasites living in the areas of malaria endemic is essential to asse... more The detailed description of parasites living in the areas of malaria endemic is essential to assess the achievement of current strategies of controlling malarial disease. The purpose of this study is to inspect the inherited multiplicity of the Plasmodium falciparum, obtained from malaria endemic areas of Pakistan, using the merozoite surface protein-2 (msp-2), a highly polymorphic gene as a genetic marker. Blood samples of Plasmodium falciparum positive patients were collected from different Basic Health Units in different provinces of Pakistan during period from January 2018 to January 2019. DNA from Plasmodium falciparum positive samples was isolated and genotyped for msp-2 using allele-speci c nested polymerase chain reaction (PCR). Total 209 isolates were collected in which 179 show positive result through PCR and msp-2 recognized in 85 (47.4%) isolates. Allele of the msp-2 gene perceived the 3D7/IC allelic family in 42.3% and FC27 allelic family typing in 57.6%. A total of 25 different msp-2 genotypes were detected in which 13 were from FC27 family and 12 were from 3D7/IC family. Overall mean multiplicity of infections (MOI) was 1.20 (95 percent CI 1.00-1.50). The msp-2 locus had a heterozygosity index of 0.5. There were no differences in MOI between age groups of study. The parasite multiplicity density of infection has negative correlation found and also signi cant (P-value = 0.02). Plasmodium falciparum isolates from Pakistan's endemic areas are mostly monoclonal, with a low degree of MOI and moderate genetic diversity in the research population.
International journal for agro veterinary and medical sciences, 2017
Mycobacterium bovis (M. bovis) is the major cause of gastrointestinal tuberculosis in humans. The... more Mycobacterium bovis (M. bovis) is the major cause of gastrointestinal tuberculosis in humans. The study was conducted in Lahore to compare 100 blood and 100 sputum samples from suspected patients of active tuberculosis, using the Duplex PCR and conventional methods for the detection of M. bovis and Mycobacterium tuberculosis (M. tuberculosis). In M. tuberculosis, pncA gene and species-specific 500-bp fragment of M. bovis was targeted in Duplex PCR. In sputum samples 37 % revealed the presence of M. tuberculosis while 05 % samples were positive for M. bovis using duplex PCR. Blood samples, 39 % and 04 % were positive for M. tuberculosis and M. bovis respectively through PCR. Acid fast bacilli (AFB) have been instituted in 23 % sputum 08 % blood samples using Ziehl-Neelsen staining. On culturing of 11 % sputum and 09 % blood samples were found positive. The sensitivity and specificity of Duplex PCR was found statistically significant in comparison to conventional methods for the differential diagnosis of tuberculosis caused by M. tuberculosis and M. bovis. Therefore Duplex PCR is a better choice of diagnostic test in the clinical setups where clinical urgencies necessitate a reliable, sensitive and specific test with results in a short time period.
Journal of Infection and Public Health
Background: Leishmaniasis is the second and fourth highest cause of mortality and morbidity respe... more Background: Leishmaniasis is the second and fourth highest cause of mortality and morbidity respectively among all tropical diseases. Recurrence in the onset of leishmaniasis is a major problem that needs to be addressed to reduce the case fatality rate and ensure timely clinical intervention. Here we are investigating the association of risk factors with recurrent cutaneous leishmaniasis to address this issue. Material and methods: Patients received by Nasser Ullah Khan Babar Hospital in Peshawar, Pakistan from March 2019 to July 2020 were enrolled in this study. Those patients who developed symptoms after completion of treatment were included in Group-A while those who had atypical scars like leishmaniasis but were negative for cutaneous leishmaniasis were included in the comparison group tagged as Group B. All those individuals who had completed six weeks of treatment for CL but had normal complete blood counts (CBC) were included to avoid other underlying immunological pathologies, while we excluded those participants who had co-morbidities like diabetes, liver disease, cardiac disease, and pregnant and lactating women through their history Association was tested between Group-A and Group-B with other explanatory variables through chi-square test. The regression model was proposed to determine the predictors. Result: A total of 48 participants of both sexes were included in the study with a mean age of 32.2 ± 15.10. The data suggest that females are overrepresented among the patients with recurrent leishmaniasis [21(53.8 %,); p = 0.07]. Compared to patients; healthy participants had a higher proportion of adults (19-59
Parasitology Research
Tick infestation is a major public and animal health concern causing significant financial losses... more Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were examined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan.
Acta Tropica, 2021
Pyrimethamine was first introduced for the treatment of malaria in Asia and Africa during the ear... more Pyrimethamine was first introduced for the treatment of malaria in Asia and Africa during the early 1980s, replacing chloroquine, and has become the first line of drugs in many countries. In recent years, development of pyrimethamine resistance in Plasmodium vivax has become a barrier to effective malaria control strategies. Here, we describe the use of meta-barcoded deep amplicon sequencing technology to assess the evolutionary origin of pyrimethamine resistance by analysing the flanking region of dihydrofolate reductase (dhfr) locus. The genetic modelling suggests that 58R and 173L single mutants and 58R/117N double mutants are present on a single lineage; suggesting a single origin of these mutations. The triple mutants (57L/58R/117N, 58R/61M/117N and 58R/117N/173L) share the lineage of 58R/117N, suggesting a common origin. In contrast, the 117N mutant is present on two separate lineages suggesting that there are multiple origins of this mutation. We characterised the allele frequency of the P. vivax dhfr locus. Our results support the view that the single mutation of 117N and double mutations of 58R/117N arise commonly, whereas the single mutation of 173L and triple mutations of 57L/58R/117N, 58R/61M/117N and 58R/117N/173L are less common. Our work will help to inform mitigation strategies for pyrimethamine resistance in P. vivax.
Pyrimethamine was first introduced for the treatment of malaria in Asia and Africa during the ear... more Pyrimethamine was first introduced for the treatment of malaria in Asia and Africa during the early 1980s, replacing chloroquine, and has become the first line of drugs in many countries. In recent years, development of pyrimethamine resistance in Plasmodium vivax has become a barrier to effective malaria control strategies. Here, we describe the use of meta-barcoded deep amplicon sequencing technology to assess the evolutionary origin of pyrimethamine resistance by analysing the flanking region of dihydrofolate reductase (dhfr) locus. The genetic modelling suggests that 58R and 173L single mutants and 58R/117N double mutants are present on a single lineage; suggesting a single origin of these mutations. The triple mutants (57L/58R/117N, 58R/61M/117N and 58R/117N/173L) share the lineage of 58R/117N, suggesting a common origin. In contrast, the 117N mutant is present on two separate lineages suggesting that there are multiple origins of this mutation. We characterised the allele freq...
Infection, Genetics and Evolution, 2020
Microscopy Research and Technique, 2020
Leishmaniasis is a worldwide public health problem and vector‐borne disease. It is caused by a di... more Leishmaniasis is a worldwide public health problem and vector‐borne disease. It is caused by a diverse group of protozoan parasites that belong to the genus Leishmania and transmitted to humans through a bite of an infected female sand fly. Leishmaniasis has attained epidemic proportion in Khyber Pakhtunkhwa and raises serious concern over its management. The present research work was conducted in cutaneous leishmaniasis (CL) prevalent village named Surgul of district Kohat, Khyber Pakhtunkhwa with a focus to investigate whether rodent can act as a source for dissemination of leishmanial species or not. In this context, rodent samples were analyzed via morphological and molecular approaches to unveil prevalence of CL. It was reported that 12.5% of samples were positive for signs of leishmaniasis through microscopy and 18.75% through polymerase chain reaction (PCR). Supporting the findings further, the color character of rodents was also taken into consideration, which shows that lig...
A total of 300 samples were collected from suspected patient to find the prevalence of Cutaneous ... more A total of 300 samples were collected from suspected patient to find the prevalence of Cutaneous Leishmaniasis as well as Polymerase Chain Reaction (PCR) based identification of respective species in the endemic areas of Dir Lower and Dir Upper (Khyber Pakhtunkhwa Province) Pakistan. These patients including (171) males and (129) females from these areas, irrespective of age, were analyzed using patient’s biopsy samples, slide smears and filter paper impressions from lesions. Kinetoplast kDNA-PCR and (Internal Transcribed Spacer 1(ITS1-PCR) were performed using RV1/RV2 and LITS R/L5.8s primers, respectively. Age group of 0-15 years was found to be the most infected with 47.5% of total cases while age group of 45 years and above showed lowest rate (7.0%). Out of 300 patients, 172 (56.17%) were positive by microscopy, 79.5% (238 Patients) by ITS1-PCR-Assay and 280 (93.5%) were confirmed by kDNA-PCR. It was concluded that cutaneous leishmaniasis in prevalent in local population of Dir ...
African Journal of Microbiology Research, 2012
A study was conducted for quantification of antibodies to haemagglutinating viruses such as AIV (... more A study was conducted for quantification of antibodies to haemagglutinating viruses such as AIV (H9N2 and H7N3), NDV and IBV in vaccinated poultry flocks. A total of 198 serum samples from different commercial broiler and layer farms in Lahore, Pakistan, were checked for their serological statuses. Haemagglutination inhibition (HI) test was performed to determine the titer of serum antibodies against the above mentioned viruses. It was observed that 25 and 44% samples were positive for H7N3 and H9N2 in broilers, respectively. The seropositivity of H7N3 and H9N2 in serum samples of commercial layers were 66 and 94%, respectively. It was recorded that 96% of the samples in broilers and 100% of those in layers, were found seropositive for antibodies against NDV. Moreover, 69 and 86% were seropositive for IBV in broilers and layers, respectively. Calculated geometric mean titers for groups of 0-2, 2-4 and 4-6 weeks of age were found to be 36.8, 78.8, and 111.4 for H9N2; 48.5, 157.6, 222.9 for NDV; and 97, 84.4, 111.4 for IBV, respectively. It was concluded that the samples were not having protective antibody titers indicating unsuccessful vaccination practiced in these poultry farms which may lead to significant economic losses.
BioMed Research International
Background. The IL-12/IFN-γ axis pathways play a vital role in the control of intracellular patho... more Background. The IL-12/IFN-γ axis pathways play a vital role in the control of intracellular pathogens such as Salmonella typhi. Objective. The study is aimed at using whole exome sequencing (WES) to screen out genetic defects in IL-12/IFN-γ axis in patients with recurrent typhoid fever. Methods. WES using next-generation sequencing was performed on a single patient diagnosed with recurrent typhoid fever. Following alignment and variant calling, exomes were screened for mutations in 25 genes that are involved in the IL-12/IFN-γ axis pathway. Each variant was assessed by using various bioinformatics mutational analysis tools such as SIFT, Polyphen2, LRT, MutationTaster, and MutationAssessor. Results. Out of 25 possible variations in the IL-12/IFN-γ axis genes, only 2 probable disease-causing mutations were identified. These variations were rare and include mutations in IL23R and ZNFX I. Other pathogenic mutations were found, but they were not considered likely to cause disease based o...
To determine the incidence of antimicrobial-resistant emerging pathogens, Clostridium difcile, an... more To determine the incidence of antimicrobial-resistant emerging pathogens, Clostridium difcile, and its associated risk factors in tertiary care setups of Pakistan. Tis cross-sectional prospective study was conducted from January 2019 to December 2020, to determine the prevalence and antimicrobial resistance patterns of C. difcile strains isolated from 450 stool specimens of patients sufering from diarrhea hospitalized in tertiary care hospitals in Peshawar, Pakistan. Te stool samples of the patients were processed for culture and detection of toxin A and toxin B by enzyme-linked immunosorbent assay (ELISA) and tpi PCR. Te drug sensitivity test was performed for antibiotics including ampicillin, cefxime, cefepime, amoxicillin, nalidixic acid, sulpha/TMP (SXT), chloramphenicol, metronidazole, vancomycin, ciprofoxacin, levofoxacin, and imipenem. Of 450 stool specimens, 108 (24%) were positive for C. difcile by stool culture, whereas 115 (25.5%) were only positive for C. difcile toxins based on ELISA and PCR (128 (28.6%). Of 108, 90.7% (n � 98) isolates were resistant to one antibiotic, and 90 (83.4%) were resistant to three or more antimicrobials. Te highest resistance rates were found against penicillin (83.3%) followed by amoxicillin (70%), nalidixic acid (61%), and metronidazole (38%), and the lowest resistance was found against vancomycin (6.4%) and imipenem (3.7%). CDI was statistically signifcantly correlated with increased age, use of antibiotics, abdominal surgeries, use of proton pump inhibitors and H2a, and presence of comorbidities. Te high frequency of C. difcile in Peshawar, Pakistan, indicates that CDI is an important nosocomial infection in diferent hospitals. Te results will be helpful for clinicians to redesign control and therapeutic strategies in hospitals.
Background. The IL-12/IFN-γ axis pathways play a vital role in the control of intracellular patho... more Background. The IL-12/IFN-γ axis pathways play a vital role in the control of intracellular pathogens such as Salmonella typhi. Objective. The study is aimed at using whole exome sequencing (WES) to screen out genetic defects in IL-12/IFN-γ axis in patients with recurrent typhoid fever. Methods. WES using next-generation sequencing was performed on a single patient diagnosed with recurrent typhoid fever. Following alignment and variant calling, exomes were screened for mutations in 25 genes that are involved in the IL-12/IFN-γ axis pathway. Each variant was assessed by using various bioinformatics mutational analysis tools such as SIFT, Polyphen2, LRT, MutationTaster, and MutationAssessor. Results. Out of 25 possible variations in the IL-12/IFN-γ axis genes, only 2 probable disease-causing mutations were identified. These variations were rare and include mutations in IL23R and ZNFX I. Other pathogenic mutations were found, but they were not considered likely to cause disease based on various mutation predictors. Conclusion. Applying WES to the patient with recurrent typhoid fever detects variants that are not much important as other genes in the IL-12/IFN-γ axis. Results of the current study suggest that a large population sizes would be needed to examine the functional relevance of IL-12/IFN-γ axis genes with recurrent typhoid fever.
Background: Leishmaniasis is the second and fourth highest cause of mortality and morbidity respe... more Background: Leishmaniasis is the second and fourth highest cause of mortality and morbidity respectively among all tropical diseases. Recurrence in the onset of leishmaniasis is a major problem that needs to be addressed to reduce the case fatality rate and ensure timely clinical intervention. Here we are investigating the association of risk factors with recurrent cutaneous leishmaniasis to address this issue. Material and methods: Patients received by Nasser Ullah Khan Babar Hospital in Peshawar, Pakistan from March 2019 to July 2020 were enrolled in this study. Those patients who developed symptoms after completion of treatment were included in Group-A while those who had atypical scars like leishmaniasis but were negative for cutaneous leishmaniasis were included in the comparison group tagged as Group B. All those individuals who had completed six weeks of treatment for CL but had normal complete blood counts (CBC) were included to avoid other underlying immunological pathologies, while we excluded those participants who had co-morbidities like diabetes, liver disease, cardiac disease, and pregnant and lactating women through their history Association was tested between Group-A and Group-B with other explanatory variables through chi-square test. The regression model was proposed to determine the predictors. Result: A total of 48 participants of both sexes were included in the study with a mean age of 32.2 ± 15.10. The data suggest that females are overrepresented among the patients with recurrent leishmaniasis [21(53.8 %,); p = 0.07]. Compared to patients; healthy participants had a higher proportion of adults (19-59
Pyrimethamine was first introduced for the treatment of malaria in Asia and Africa during the ear... more Pyrimethamine was first introduced for the treatment of malaria in Asia and Africa during the early 1980s, replacing chloroquine, and has become the first line of drugs in many countries. In recent years, development of pyrimethamine resistance in Plasmodium vivax has become a barrier to effective malaria control strategies. Here, we describe the use of meta-barcoded deep amplicon sequencing technology to assess the evolutionary origin of pyrimethamine resistance by analysing the flanking region of dihydrofolate reductase (dhfr) locus. The genetic modelling suggests that 58R and 173L single mutants and 58R/117N double mutants are present on a single lineage; suggesting a single origin of these mutations. The triple mutants (57L/58R/117N, 58R/61M/117N and 58R/117N/173L) share the lineage of 58R/117N, suggesting a common origin. In contrast, the 117N mutant is present on two separate lineages suggesting that there are multiple origins of this mutation. We characterised the allele frequency of the P. vivax dhfr locus. Our results support the view that the single mutation of 117N and double mutations of 58R/117N arise commonly, whereas the single mutation of 173L and triple mutations of 57L/58R/117N, 58R/61M/117N and 58R/117N/173L are less common. Our work will help to inform mitigation strategies for pyrimethamine resistance in P. vivax.
Objectives: Rabbit coccidiosis is caused by Eimeria sp. inhabiting the liver and intestine. The p... more Objectives: Rabbit coccidiosis is caused by Eimeria sp. inhabiting the liver and intestine. The present study investigated the coccidiosis in rabbits in terms of age, humidity, sex variations and temperature and chemotherapy was used in the form of allopathic and herbal drugs. Methods: A total of 112 rabbits possibly infected during coccidiosis outbreak were obtained from different locations of Lahore, Pakistan. The faecal examination was performed using direct smear, floatation to observe the oocyst of Eimeria spp. Results: It was observed that females were slightly more susceptible (47.2 %) than males (44.7 %); also the incidence of coccidiosis was more prevalent in baby kits from 1-3 months (25.3 %) as compared to adults (15.2 %). Month-wise analysis depicted that, the infection was more prevalent in the month of March, and July (58.3 %, 60.7 %, and 73.9 % respectively due to high humidity. Coccidiosis also changes in blood parameters with increased level of WBCs (20.8 %), neutrophils (17.5 %), monocytes (29.1 %) and basophils (11.3 %) while decreases RBCs (44.9 %), lymphocytes (18.3 %) and haemoglobin level (35.4 %). Chemical drugs minimize OPG (Oocyst per Gram) dramatically such as Sulphadimidine sodium and toltrazuril efficiently decreases the OPG to 48 % and 74.4 %, respectively. Herbal drugs in the form of Nigella sativa L. seeds (kalonji) minimized the OPG gradually 57 % and 53.4 % oocysts respectively in 14 days. Garlic extract reduces 57.8 % oocytes in faeces. Conclusions: The chemotherapy suggested that toltrazuril is more efficient for quick action compared to Sulphadimidine Sodium. In addition, we found that garlic and black seeds were more effective in reducing the OPG level.
Leishmaniasis is a worldwide public health problem and vector-borne disease. It is caused by a di... more Leishmaniasis is a worldwide public health problem and vector-borne disease. It is caused by a diverse group of protozoan parasites that belong to the genus Leishmania and transmitted to humans through a bite of an infected female sand fly. Leishmaniasis has attained epidemic proportion in Khyber Pakhtunkhwa and raises serious con
Background. Antibiotic supplementation in feed has been continued for the previous 60 years as th... more Background. Antibiotic supplementation in feed has been continued for the previous 60 years as therapeutic use. ey can improve the growth performance and feed efficiency in the chicken flock. A favorable production scenario could favor intestinal microbiota interacting with antibiotic growth promoters and alter the gut bacterial composition. Antibiotic growth promoters did not show any beneficial effect on intestinal microbes. Scope and Approach. Suitable and direct influence of growth promoters are owed to antimicrobial activities that reduce the conflict between host and intestinal microbes. Unnecessary use of antibiotics leads to resistance in microbes, and moreover, the genes can relocate to microbes including Campylobacter and Salmonella, resulting in a great risk of food poisoning. Key Findings and Conclusions. is is a reason to find alternative dietary supplements that can facilitate production, growth performance, favorable pH, and modulate gut microbial function. erefore, this review focus on different nutritional components and immune genes used in the poultry industry to replace antibiotics, their influence on the intestinal microbiota, and how to facilitate intestinal immunity to overcome antibiotic resistance in chicken.
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Papers by Dr mubashir Hussain