Muhammad Hassan Mushtaq

Complete genome analysis of a highly pathogenic H5N1 influenza A virus isolated from a tiger in China.

An influenza A virus (A/Tig/SH/01/2005 (H5N1) was isolated from lung tissue samples of a dead zoo tiger with respiratory disease in China in July 2005. Complete genome analysis indicated that the isolate was highly identical to an H5N1 virus isolated from a migratory duck at Poyang lake in China in that year. The genotype of the isolate was K,G,D,5J,F,1J,F,1E, and phylogenetically it was a clade 2.2 virus. Molecular characterization of all of the gene segments revealed characteristics of highly pathogenic influenza A viruses. These results may help to identify molecular determinants of virulence and highlight the necessity for continuous surveillance.

Protective immune responses induced by in ovo immunization with recombinant adenoviruses expressing spike (S1) glycoprotein of infectious bronchitis virus fused/co-administered with granulocyte-macrophage colony stimulating factor

Infectious bronchitis virus (IBV) causes tremendous economic losses associated with production inefficiencies and mortality in poultry industry worldwide. In the present report, the recombinant adenoviruses expressing chicken granulocyte-macrophage colony stimulating factor (GM-CSF) and S1 gene of nephropathogenic IBV were constructed and characterized. Then, the immunological efficacy and protection against homologous IBV challenge were assessed in specific pathogen free (SPF) chickens. The results showed that the chickens vaccinated in ovo with rAd-S1, rAd-GM-S1 (GM-CSF fused with S1 using glycine linkers) and rAd-GM-CSF plus rAd-S1 (co-administered) developed specific anti-IBV HI antibodies. Moreover, the fusion of the GM-CSF markedly increased spleen cell proliferation and IFN-γ production while mild increased in IL-4 production, which demonstrated the enhancement of cell-mediated immune responses. Following challenge with IBV, the chickens in the group vaccinated with rAd-S1 fused or co-administered with GM-CSF had fewer nephropathic lesions and showed 100% protection as compared to that of rAd-S1 alone which showed 70% protection. It indicated that the single dose in ovo vaccination of the GM-CSF fused or co-administered with S1 of IBV could enhance significantly the humoral, cellular immune responses and provide complete protection against nephropathogenic IBV challenge. This finding may provide basic information for effective in ovo vaccines design against IBV.

Anthelmintic activity of Artemisia vestita Wall ex DC. and Artemisia maritima L. against Haemonchus contortus from sheep.

Current study was designed to evaluate in vivo and in vitro anthelmintic activity of Artemisia vestita Wall ex DC. and Artemisia maritima L. against Haemonchus contortus in comparison with ivermectin to investigate the effect of plant extracts on survival of infective L3 and adults under in vitro condition. Plant extracts were given to H. contortus infected sheep orally and it was infected with L3 stage of H. contortus at dose of 5000 larvae/sheep. Total of 25-30 larvae were incubated with plant extracts in PBS alone and ivermectin at different concentration used as positive control. It was recorded that there is a significant decrease in fecal egg count (FEC) after post-treatment period for both plants. The highest fecal egg count reduction for A. vestita was 87.2% at 100mg/kg while for A. maritima it was 84.5% on day 28 post-treatment. Investigated extracts indicated significant activity against larvae and adult worms.

Prevalence and genetic characterization of avian polyomavirus and psittacine beak and feather disease virus isolated from budgerigars in Mainland China

Budgerigar fledgling disease (BFD) and psittacine beak and feather disease (PBFD) are caused by avian polyomavirus (APV) and psittacine beak and feather disease virus (PBFDV), respectively. These diseases frequently infect psittacine birds and result in similar clinical manifestations. In this study, we observed the prevalence of PBFDV infection and a dual infection of APV and PBFDV in a budgerigar (Melopsittacus undulatus) in Mainland China for the first time. One PBFDV isolate and two APV isolates were harvested using chicken embryos. Genetic characterization and phylogenetic analysis of the complete genome of the two APV isolates revealed nucleotide similarity ranging from 99.0% to 99.6% to other sequences in GenBank, and a 14-bp insertion was observed in the genome of one APV isolate. The results of complete genome analysis of the PBFDV isolate showed nucleotide similarity ranging from 83.0% to 95.0% with other PBFDV sequences in GenBank. Genetic characterization and phylogenetic analysis of the APV and PBFDV strains isolated in this study indicated that the isolates from China were closely related to their Japanese counterparts. The results of this study will help to identify molecular determinants and will aid further research on the prevention and control of APV and PBFD infection.

Zoonotic tuberculosis in occupationally exposed groups in Pakistan

BACKGROUND Bovine tuberculosis (bTB) remains a public health threat in low-income countries. Data on human tuberculosis (TB) due to Mycobacterium bovis are poorly documented in developing countries. AIMS To determine the occurrence of active pulmonary TB due to M. bovis in abattoir workers, butchers, livestock farmers and veterinarians and to document their knowledge and practices regarding bTB. METHODS A cross-sectional study of abattoir workers, butchers, livestock farmers, veterinary doctors and veterinary assistants, which obtained data on socio-demographic conditions, knowledge and practices regarding TB. Sputum samples were collected from respondents with a chronic cough. The chi-squared test was used for statistical analysis. RESULTS A total of 141 abattoir workers, 317 butchers, 50 livestock farmers, five veterinary doctors and three veterinary assistants took part. Four out of 16 abattoir workers and 1 out of 50 livestock farmers were positive for M. bovis by PCR analysis. Duration of work as an abattoir worker was significantly associated (P < 0.05) with prevalence of zoonotic TB. The knowledge of abattoir workers, butchers, livestock farmers and veterinary assistants regarding transmission of bTB from animals to humans and the symptoms of TB in humans was very poor. Most of these workers did not use protective equipment and appropriate safe working techniques and were considered at high risk of acquiring zoonotic TB. CONCLUSIONS Zoonotic TB is a significant public health issue among professionally exposed groups in Peshawar, Pakistan and suggests a need for further detailed investigations of the disease in this and similar areas.

Aspirin may do wonders by the induction of immunological self-tolerance against autoimmune atherosclerosis

Induction of immune tolerance is one of the recent novel immunomodulatory strategies to directly intervene the autoimmune-driven atherosclerosis. Aspirin is a prototypic non-steroidal anti-inflammatory drug, which is now being regarded as a life-saver in variety of atherosclerotic cardiovascular complications. Considerable amount of data emerged during last few years clearly suggests that aspirin can cause immunomodulation by several mechanisms, particularly, its ability to induce tolerogenic dendritic cells (DCs) and to upregulate T regulatory (Treg) cells is especially appealing with respect to induction of immunological self-tolerance. Based on this fact, we hypothesize that aspirin, in addition to its anti-inflammatory effect, may also specifically inhibit autoimmune response in atherosclerosis by actively increasing CD4+CD25+FOXP3+Treg cells as well as by inducing tolerogenic DCs which induce hyporesponsiveness in responder naïve T cells. If proved to be correct, this hypothesis will provide an opportunity to medical community with an already available aspirin-based immunotherapeutic approach for inducing immune tolerance against atherosclerosis.

Molecular Epidemiology of a novel re-assorted epidemic strain of equine influenza virus in Pakistan in 2015–16

BACKGROUND A widespread epidemic of equine influenza (EI) occurred in nonvaccinated equine population across multiple districts in Khyber Pakhtunkhwa Province of Pakistan during 2015-2016. OBJECTIVES AND METHODS An epidemiological surveillance study was conducted from Oct 2015 to April 2016 to investigate the outbreak. EI virus strains were isolated in embryonated eggs from suspected equines swab samples and were subjected to genome sequencing using M13 tagged segment specific primers. Phylogenetic analyses of the nucleotide sequences were concluded using Geneious. Haemagglutinin (HA), Neuraminidase (NA), Matrix (M) and nucleoprotein (NP) genes nucleotide and amino acid sequences of the isolated viruses were aligned with those of OIE recommended, FC-1, FC-2, and contemporary isolates of influenza A viruses from other species. RESULTS HA and NA genes amino acid sequences were very similar to Tennessee/14 and Malaysia/15 of FC-1 and clustered with the contemporary isolates recently reported in the USA. Phylogenetic analysis showed that these viruses were mostly identical (with 99.6% and 97.4% nucleotide homology) to, and were reassortants containing chicken/Pakistan/14 (H7N3) and Canine/Beijing/10 (H3N2) like M and NP genes. Genetic analysis indicated that A/equine/Pakistan/16 viruses were most probably the result of several re-assortments between the co-circulating avian and equine viruses, and were genetically unlike the other equine viruses due to the presence of H7N3 or H3N2 like M and NP genes. CONCLUSION Epidemiological data analysis indicated the potential chance of mixed, and management such as mixed farming system by keeping equine, canine and backyard poultry together in confined premises as the greater risk factors responsible for the re-assortments. Other factors might have contributed to the spread of the epidemic, including low awareness level, poor control of equine movements, and absence of border control disease strategies.

Some challenges in progressive control of livestock originated zoonotic diseases in Pakistan-a pilot survey

ABSTRACT Objective To document the recent livestock related practices and possible unhygienic ways of pathogen entry. Identification of the potential risk factors for the spread of infection is important to design an evidence-based disease control programme. Methods Rapid assessment method was adopted and a purposive sample of 60 dairy farmers were interviewed. The following factors were noted for contributing in primary and secondary transmission of zoonotic infections: (i) persons who come in close contact with animals and their secretions, (ii) management strategies of farm animals (sheds and environment), (iii) management practices adopted at farms, (iv) small scale farmers and rural livestock production systems, (v) milk collection systems. Results This research unveiled the certain routes of zoonotic disease transmission. Certain management practices, precautionary measures and strategies were the pivotal risk factors. Conclusions The study emphasizes the need to educate the poor livestock keepers.

Comparative efficacy of different antidotes against experimental nitrate intoxication in rabbits

&NA; The antidotal efficacy of aqueous garlic extract, methylene blue, and velenium (Vitamin E + sodium selenite) was compared against experimental nitrate intoxication in rabbits. Forty‐two, albino rabbits of identical age, gender, and body weight were randomly divided into 7 groups (A to G) and subjected to experimental treatments for a period of 40 days. Rabbits of group A were offered only normal feed and served as negative control, while, rabbits of group B constituted the positive control group and received feed supplemented with sodium nitrate at 400 mg/kg body weight. Sodium nitrate‐containing feed and intraperitoneal injection of 1% methylene blue solution at 2 mg/kg body weight were administered to group C. Rabbits of group D were given sodium nitrate‐supplemented feed and aqueous garlic extract at 500 mg/kg body weight through oral route. Group E was treated with sodium nitrate‐added feed, intraperitoneal injection of 1% methylene blue solution at 2 mg/kg body weight, and oral administration of garlic extract at 500 mg/kg. Velenium (25 mg of Vitamin E + 2.2 mg of sodium selenite per ml) was intraperitoneally injected at 1 ml/kg body weight to rabbits of group F along with the provision of sodium nitrate‐supplemented feed. In addition to being fed with sodium nitrate‐containing feed, group G obtained intraperitoneal injection of velenium at 1 ml/kg body weight and oral administration of garlic extract at 500 mg/kg body weight. The efficacy of antidotes was assessed on the basis of changes in blood nitrite level, biochemical profile, and gross pathological lesions manifested by the treated rabbits. The combination of methylene blue and garlic extract was highly effective in treating nitrate toxicity followed by methylene blue, garlic extract, and velenium, respectively. Whereas, the concurrent administration of garlic extract and velenium was least efficacious in terms of antidotal efficacy. In conclusion, aqueous garlic extract can be effectively used either alone or in combination with methylene blue when treating rabbits diagnosed with nitrate toxicity.

Investigating the epidemiology of EI epidemic spread in the Province of Khyber Pakhtunkhwa, Pakistan in 2015–16

EI in non-vaccinated population causes disruption and economic losses. To identify the risk factors associated with the EI epidemics in equids in Pakistan, a 1:1 matched case control study was conducted during 2015-2016. Including a total of 197 laboratory confirmed cases and negative controls, matched on the basis of geography, time of sampling, specie and age. A piloted questionnaire was used to collect data regarding risk factors associated with the occurrence of EI in face to face interviews. Conditional logistic regression was performed to analyze the data. A total of 16 out of 23 variables were found associated as risk factors in Univariable conditional logistic regression analysis. Multivariable conditional logistic-regression analysis was also performed. Monthly removal of manure doubles the risk of EI (EI) compared to its daily removal. Due to lack of vaccination; the spread of disease was favored by high equine density. Investigating the index-case it was recorded that infected cases were imported from Afghanistan. Most of these risk factors related to biosecurity and management were due to low awareness level regarding EI amongst the respondents. These findings are in line with the results of many other studies identifying similar risk factors for EI infection in various countries. Adopting protective practices, vaccination and controlling the risk factors identified in the present study could reduce the spread and future outbreaks of EI in Pakistan.