Identification of provenance of Basmati rice grown in different regions of Punjab through multivariate analysis.

High-priced Basmati rice is vulnerable to deliberate mislabeling to increase profits. This type of fraud may lower consumers' confidence as inferior products can affect brand reputation. To address this problem, there is a need to devise a method that can efficiently distinguish Basmati rice grown in regions that are famous versus the regions that are not suitable for their production. Therefore, in this investigation, thirty-six samples of Basmati rice were collected from two zones of Punjab province (one known for Basmati rice) of Pakistan which is the major producer of Basmati rice. The elemental composition of rice samples was assessed using inductively coupled plasma-optical emission spectrometry and an organic elemental analyzer, whereas data on δ13C was acquired using isotopic ratio-mass spectrometry. Regional clustering of samples based on their respective cultivation zones was observed using multivariate data analysis techniques. Partial least squares-discriminant analysis was found to be effective in grouping rice samples from the different locations and identifying unknown samples belonging to these two regions. Further recommendations are presented to develop a better model for tracing the origin of unidentified rice samples.

GAMMA SPECTROMETRY AND NEUTRON ACTIVATION ANALYSIS OF TOOT OILFIELD WELLS OF DATTA FORMATION, UPPER INDUS BASIN, DISTRICT ATTOCK, PUNJAB, PAKISTAN.

Present study was aimed to investigate trace elemental composition and possible elemental correlation with depth of some Pakistani wells. Exploratory oil wells of Toot oilfield from Potwar region of upper Indus basin were selected for this purpose. Samples of well cuttings and soil sections were collected with the assistance of Oil and Gas Development Company Limited. Elemental analyses were performed using instrumental neutron activation analysis. This study is the first of its kind to report detailed elemental profile of Toot oilfield, Attock, Punjab, Pakistan, via instrumental neutron activation analysis (INAA). Source formation of Toot oilfield is characterized by Jurassic Datta formation. Around 19-26 elements were identified by INAA. Major, minor and trace elemental concentrations varied with depths along the sampling lines. Data analysis was performed by the application of principal component analysis (PCA). PCA was useful in differentiating between surface and depth samples. Elemental volumes of different wells were established through bi-plots. In extension, naturally occurring radioactive materials and technologically enhanced naturally occurring radioactive materials were also determined for the wells of Toot oilfield. Annual dose rates and activity levels were calculated in oil well formations. Measurements of annual dose rates fall within safe limits, indicating anodyne atmosphere. Contamination indices, such as enrichment factor with respect to Al, geo accumulation index (Igeo), pollution index and integrated pollution index, were also determined. Cr and Zn were found to cross the safe limits, which may be due to the local industrial and anthropogenic activities.

Physiological properties of indigenous lytic bacteriophages as monophage suspension and cocktail against poultry-adapted typhoidal Salmonella variants.

The emergence and spread of multidrug resistance among pathogens of the agro-food sector is increasing at an alarming rate, which has directed attention to the search for alternative to antibiotic therapy. The present work studied the physiological and population dynamics of lytic bacteriophages against avian-adapted Salmonella. Out of 28 positive samples, four bacteriophage isolates (SalØ-ABF37, SalØ-RCMPF12, SalØ-MCOH26, SalØ-DNLS42) were selected based on their ability to clearly lyse bacterial test strains. The isolates propagated were active against closely related D1 serotypes, i.e., S. Enteritidis and S. Typhimurium, with no heterologous activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 23235. Each of the monophage suspension and cocktail efficiently suppressed the bacterial count from exponential increase in comparison to the untreated bacterial control. The bacterial turbidity was recorded as 0.244 at λ600 during 400 min of co-incubation, in contrast to bacterial control showing λ600 = 0.669. The latent period was recorded to be 25, 35, 25 and 30 for SalØ-ABF37, SalØ-RCMPF12, SalØ-MCOH26 and SalØ-DNLS42, with 73.00, 97.00, 132 and 75.00 PFU cell-1, respectively. The highest lytic activity was seen at 37.00 ˚C - 42.00 ˚C, with phage particle count being fairly stable at pH 3.00 - 9.00. Each of the isolates possessed dsDNA by being resistant to RNase A. The current study concludes that lytic phages are promising alternative to combat multidrug resistant superbugs. The physiological characterization and bacterial growth inhibition are important parameters in standardization of phage therapy.

Quantification and Trends of Antimicrobial Use in Commercial Broiler Chicken Production in Pakistan.

Antimicrobial resistance (AMR) is a global health challenge and antimicrobial use (AMU) in the livestock sector has been considered as one of the contributing factors towards the development of AMR in bacteria. This study summarizes the results of a point prevalence survey conducted to monitor farm-level AMU in commercial broiler chicken farms in Punjab and Khyber Pakhtunkhwa (KPK) provinces of Pakistan. A cross-sectional study was conducted to quantify AMU and to check seasonal variations of AMU in 12 commercial broiler chicken farms (six from each province) during the summer and winter seasons of the year 2020-2021. AMU was recorded using three AMU metrics: kg, mg per population correction unit (mg/PCU), and mg/kg of final flock weight. A total of 22 antimicrobial drugs (348.59 kg) were used for therapeutic or prophylactic purposes in surveyed broiler chicken farms. The total combined AMU for all the broiler chicken farms was 462.57 mg/PCU. The use of most of the antimicrobials increased during winter flocks compared to summer. The top three antimicrobial drugs used during the summer were neomycin (111.39 mg/PCU), doxycycline (91.91 mg/PCU), and tilmicosin (77.22 mg/PCU), whereas doxycycline (196.81 mg/PCU), neomycin (136.74 mg/PCU), and amoxicillin (115.04 mg/PCU) during the winter. Overall, 60% of the antibiotics used in broiler chicken were critically important antimicrobial classes (CIA) for human medicine as characterized by the World Health Organization. Our findings showed high AMU in broiler chicken production and a call for urgent actions to regulate CIA use in food animals in Pakistan. This baseline survey is critical for the design and implementation of a subsequent national level AMU surveys that can include additional farming types, animals' species, and geographical locations over a longer period of time.

Bacillus spp. Contamination: A Novel Risk Originated From Animal Feed to Human Food Chains in South-Eastern Bangladesh.

The current study provides information on Bacillus spp. contamination along with present status in commercially available poultry and animal feeds as well as animal-derived products in Bangladesh. The research has been conducted to determine if animal feed and its components are a source of Bacillus spp. contamination in feed and food chain. Out of 180 different feeds, milk, egg, and human stool samples, 218 Bacillus spp. were isolated and identified by cultural morphology, microscopic, biochemical, and molecular characteristics where B. cereus, B. subtilis, B. amyloliquefaciens, B. licheniformis, B. thuringiensis, B. megaterium, and B. coagulans accounted for 51, 22, 9.1, 5.9, 5, 3.6, and 2.2%, respectively. Regarding the enumeration of total viable count and total Bacillus count, correspondingly 67 and 39% samples were found to be contaminated with above 10,000 CFU/g, while highest contamination was 85 and 75% in broiler feed, respectively. The total number of bacteria above the regulatory limits in commercially available feeds indicates a poor compliance with regulation and abuse administration in the Bangladeshi market. Moreover, a hospital-based survey showed that food-borne Bacillus spp. contributed to 4.5% human diarrhea cases and 25% food contamination associated with vegetables, rice, RTE food, milk, and egg, accounting for 46, 34, 14, 4, and 2%, respectively. B. cereus was the dominant isolate correspondingly accounting for 56 and 51% egg and milk contamination followed by B. amyloliquefaciens (32%) and B. thuringiensis (12%) in egg and B. subtilis (25%), B. amyloliquefaciens (12%), B. thuringiensis (6.4%), and B. coagulans (3.2%) in milk, respectively. Toxin gene profiling of Bacillus spp. revealed that B. cereus constituted a principal part of virulence, while B. thuringiensis, B. licheniformis, B. megaterium, B. coagulans, and B. subtilis showed genetic diversity and B. amyloliquefaciens had not carried any toxin gene. Detection rate of enterotoxin genes (nheA, nheB, nheC, cytK, hblA, hblC, hblD, and entFM) showed that 55% isolates carried nheABC genes, 80% entFM, and 71% cytK, whereas only 33% of the isolates contained hblACD gene clusters. These virulence genes were posing a threat to human health due to spread across the food and feed chain. Finally, our findings support the hypothesis that B. cereus might contribute to clinical diarrhea, gizzard erosion, and lung infection in duck and poultry, and that it contaminates animal-derived foods resulting in toxicity and antibacterial resistance to humans. Therefore, maximal tolerance limits of Bacillus spp. and their potential risks to the animal industry are urgently needed to clarify. Moreover, Bacillus spp.-induced toxin residual must be altered for human health via food chain transmission.

Pulmonary endothelium-derived PD-L1 induced by the H9N2 avian influenza virus inhibits the immune response of T cells.

BACKGROUND: The PD-1/PD-L1 pathway is an inhibitory signaling pathway that maintains the balance between the immune response and immunotolerance, and its overactivation in cancer and viral infections inhibits T cell function. The target cells of various viruses, microvascular endothelial cells (MECs) have been shown to be key regulatory points in immune regulation and virion diffusion in vivo during infection with multiple influenza virus subtypes. Furthermore, avian influenza virus (AIV) infection can induce immunosuppression by causing imbalances in immune responses and immune organ damage. Thus, the aim of this study was to investigate whether the H9N2 virus inhibited the immune function of T cells that migrated across MECs by upregulating PD-L1 expression on MECs. METHODS: The susceptibility of rat pulmonary microvascular endothelial cells (RPMECs) to the H9N2 virus was evaluated by a plaque-forming assay and immunofluorescence staining. Then, we quantified the mRNA and protein levels of PD-L1 in RPMECs induced by H9N2 virus infection using quantitative real-time PCR and flow cytometry. The interaction between the activated T cells and RPMECs infected with the H9N2 virus was revealed using a coculture system. The effect of endothelial-derived PD-L1 on T cell function was investigated by using ELISA and flow cytometry with or without a PD-L1-specific antibody. RESULTS: Surface staining and the plaque-forming assay showed that the H9N2 virus infected and replicated in RPMECs. Both the PD-L1 mRNA level and PD-L1 protein level were upregulated in RPMECs infected with the H9N2 virus. H9N2 virus-induced PD-L1 expression significantly reduced the secretions of IL-2, IFN-γ and granzyme B and perforin expression in T cells. The above data were significantly increased after treatment with an anti-PD-L1 antibody, confirming the above mentioned findings. In addition, the induction of PD-L1 expression decreased the proliferative capacity of the cocultured T cells but did not affect the apoptosis rate of T cells. CONCLUSIONS: Taken together, the results suggest that the H9N2 virus is able to inhibit the T cell immune response by upregulating PD-L1 expression in pulmonary microvascular endothelial cells.

Correction: miR29a and miR378b Influence CpG-Stimulated Dendritic Cells and Regulate cGAS/STING Pathway.

The authors wish to make the following corrections to this paper [...].

miR29a and miR378b Influence CpG-Stimulated Dendritic Cells and Regulate cGAS/STING Pathway.

The Cytosine-phosphate-guanosine (CpG) motif, which is specifically recognized intracellularly by dendritic cells (DCs), plays a crucial role in regulating the innate immune response. MicroRNAs (miRNAs) can strongly influence the antigen-presenting ability of DCs. In this study, we examine the action of miRNAs on CpG-stimulated and control DCs, as well as their effect on cyclic guanosine monophosphate-adenosine monophosphate (GMP-AMP) synthase (cGAS) and the stimulator of interferon genes (STING) signal pathway. Firstly, we selected miRNAs (miR-29a and miR-378b) based on expression in CpG-stimulated mouse bone marrow-derived dendritic cells (BMDCs). Secondly, we investigated the functions of miR-29a and miR-378b on CpG-stimulated and unstimulated BMDCs. The results showed that miR-29a and miR-378b increased expression of both the immunoregulatory DC surface markers (CD86 and CD40) and the immunosuppressive molecule CD273 by DCs. Thirdly, cytokine detection revealed that both miR-29a and miR-378b enhanced interferon-ß (IFN-ß) expression while suppressing tumor necrosis factor-α (TNF-α) production. Finally, our results suggest that miR-378b can bind TANK-binding kinase binding protein 1 (TBKBP1) to activate the cGAS/STING signaling pathway. By contrast, miR-29a targeted interferon regulatory factor 7 (IRF7) and promoted the expression of STING. Together, our results provide insight into the molecular mechanism of miRNA induction by CpG to regulate DC function.

Elemental Characterization of Medicinal Plants and Soils from Hazarganji Chiltan National Park and Nearby Unprotected Areas of Balochistan, Pakistan.

The aim of this work was to evaluate the variability in elemental composition of seven medicinal plants and their respective soils belonging to protected and nearby unprotected sites of the Hazarganji Chiltan National Park. The medical plants under study were Achillea wilhelmsii C. Koch, Peganum harmala Linn, Sophora mollis (Royle) Baker, Perovskia atriplicifolia Benth, Seriphidium quettense (Podlech.) Ling, Hertia intermedia (Bioss) O. Ktze, and Nepeta praetervisa Rech. F. Macro (C, H, N, S, K, Ca), micro (Cl, Cu, Fe, Mn, and Zn), beneficial (Al, Co, Na), others (As, Br, Cr, Cs, Hf, Pb, Rb, Sb, Sr, Sn, V and Th) and rare earth elements (Ce, Eu, La, Lu, Nd Sc, Sm, Tb and Yb) were characterized by means of standard organic elemental and instrumental neutron activation methodologies and by flame atomic absorption spectroscopy. Results showed that, among macro nutrients, carbon concentration was the highest element in both plant and soil samples followed by H and K. Elements such as Cl, Na and Fe were detected in considerably good amounts; all the other elements were found in trace quantities. Principal component analysis (PCA) was applied to identify spatial variation in elemental composition of medicinal plants, in which 80-90% of the total variance in whole set of data was found. In particular, the findings highlighted the presence of essential and beneficial elements such as C, H, N, K, Ca, Fe, Mn and Na, in samples from protected sites, while potentially dangerous elements such as Al, As, Br and Cr were detected in samples from unprotected sites. These results emphasized on the need for rational exploitation of valuable medicinal plants and supporting protected areas as an excellent source of biodiversity conservation.

Genomic Characterization of Antibiotic Resistant Escherichia coli Isolated From Domestic Chickens in Pakistan.

Poultry husbandry is important for the economic health of Pakistan, but the Pakistani poultry industry is negatively impacted by infections from Escherichia coli. We performed Illumina whole genome sequencing on 92 E. coli isolates obtained from the livers of deceased chickens originating in five Pakistani geographical regions. Our analysis indicates that the isolates are predominantly from the B1 and A clade and harbor a diverse number of antibiotic resistance and virulence genes, with no linkage between phylogeny and antibiotic resistance gene presence but some association between phylogeny and virulence gene and SNP presence for the B1 and E phylogroups. The colistin resistance gene mcr-1 and the quinolone resistance gene qnrS1 were both found in 13/92 isolates. Alarmingly, 82/92 of the E. coli strains characterized in this study are multidrug resistant with 100% (92/92) resistance to lincomycin, 81.5% (75/92) to streptomycin, 79.3% (73/92) to ampicillin and 66.3% (61/92) to ciprofloxacin. These results provide a high-resolution analysis of poultry-associated E. coli isolates in an area with a high endemic burden of antibiotic resistance. Surveillance of antibiotic resistance in poultry associated E. coli isolates is an important pillar of the One Health concept to integrate analysis of potential pathogens in human, animal, and environmental niches.

Occurrence and profile of organophosphorus compounds in fine and coarse particulate matter from two urban areas of China and Pakistan.

Seven organophosphorus compounds (OPs) were measured in urban fine and coarse particulate matter (PM) collected from two sites of Nanjing (XCNUC), China and Islamabad (APHSP), Pakistan. The fine PM mass at APHSP site was significantly higher (p = 0.005) in the spring (mean 22.5 µg/m3) than in the summer (mean 12.7 µg/m3). The total concentration, ∑7OPs, of samples collected at APHSP was found significantly higher in coarse (range 672-47621 pg/m3) than in fine PM (range 1200-15213 pg/m3); while ∑7OPs from XCNUC in fine (range 1696-15063 pg/m3) and coarse PM (range 2053-5379 pg/m3) were statistically different in samples during summer, based on two-sample t-test at 0.05 confidence interval. Seasonally, ∑7OPs was found to be higher in the samples of Nanjing (9510 ± 3633 pg/m3) in the summer than in the spring. In contrast, the samples of Islamabad had higher ∑7OPs (25558 ± 16255 pg/m3) in the spring than in the summer. Tri(chloropropyl) phosphate (TCEP) was found at higher concentration than any other OPs in the samples from the XCNUC site. Triphenyl phosphate (TPhP) was found at extremely high levels from APHSP, which may attributed to its use in jet fuel. Most of the ∑OPs concentrations were found significantly different in both PMs at both sites due to their diverse sources. Both sites showed strong correlation of ∑OPs with TCEP in both PMs, indicating similar mode of transfer from sources like plastics, lacquer, paint, glue, industrial processes and foam fillers. Relative abundance of OPs in fine and coarse PM differs amongst OP congeners and concerning seasons, with these variations attributed to different mechanisms of mass transfer such as volatilization, condensation and abrasions. Moreover, triphenylphosphine oxide (TPPO) has been reported in outdoor PMs. This is the first study on the occurrence of OPs in atmospheric fine and coarse PM and their seasonal variation from Pakistan and China.

Source apportionment using reconstructed mass calculations.

A long-term study was undertaken to investigate the air quality of the Islamabad/Rawalpindi area. In this regard fine and coarse particulate matter were collected from 4 sites in the Islamabad/Rawalpindi region from 1998 to 2010 using Gent samplers and polycarbonate filters and analyzed for their elemental composition using the techniques of Neutron Activation Analysis (NAA), Proton Induced X-ray Emission/Proton Induced Gamma-ray Emission (PIXE/PIGE) and X-ray Fluorescence (XRF) Spectroscopy. The elemental data along with the gravimetric measurements and black carbon (BC) results obtained by reflectance measurement were used to approximate or reconstruct the particulate mass (RCM) by estimation of pseudo sources such as soil, smoke, sea salt, sulfate and black carbon or soot. This simple analysis shows that if the analytical technique used does not measure important major elements then the data will not be representative of the sample composition and cannot be further utilized for source apportionment studies or to perform transboundary analysis. In this regard PIXE/PIGE and XRF techniques that can provide elemental compositional data for most of the major environmentally important elements appear to be more useful as compared to NAA. Therefore %RCM calculations for such datasets can be used as a quality assurance (QA) measure to treat data prior to application of chemometrical tools such as factor analysis (FA) or cluster analysis (CA).

Sequence and phylogenetic analysis of virulent Newcastle disease virus isolates from Pakistan during 2009-2013 reveals circulation of new sub genotype.

Despite observing the standard bio-security measures at commercial poultry farms and extensive use of Newcastle disease vaccines, a new genotype VII-f of Newcastle disease virus (NDV) got introduced in Pakistan during 2011. In this regard 300 ND outbreaks recorded so far have resulted into huge losses of approximately USD 200 million during 2011-2013. A total of 33 NDV isolates recovered during 2009-2013 throughout Pakistan were characterized biologically and phylogenetically. The phylogenetic analysis revealed a new velogenic sub genotype VII-f circulating in commercial and domestic poultry along with the earlier reported sub genotype VII-b. Partial sequencing of Fusion gene revealed two types of cleavage site motifs; lentogenic (112)GRQGRL(117) and velogenic (112)RRQKRF(117) along with some point mutations indicative of genetic diversity. We report here a new sub genotype of virulent NDV circulating in commercial and backyard poultry in Pakistan and provide evidence for the possible genetic diversity which may be causing new NDV out breaks.

Sequence and phylogenetic analysis of highly pathogenic avian influenza H5N1 viruses isolated during 2006-2008 outbreaks in Pakistan reveals genetic diversity.

BACKGROUND: Since the first outbreak recorded in northern areas of Pakistan in early 2006, highly pathogenic avian influenza H5N1 viruses were isolated from commercial poultry and wild/domestic birds from different areas of Pakistan up to July 2008. Different isolates of H5N1 were sequenced to explore the genetic diversity of these viruses. RESULTS: Phylogenetic analysis revealed close clustering and highest sequence identity in all 8 genes to HPAI H5N1 isolates belonging to unified H5 clade 2.2, sub-lineage EMA-3 recovered from Afghanistan during the same time period. Two subgroups within Pakistani H5N1 viruses, from domestic and wild birds, were observed on the basis of their sequence homology and mutations. HPAI motif, preferred receptor specificity for α-(2, 3) linkages, potential N-linked glycosylation sites and an additional glycosylation site at the globular head of HA protein of four Pakistani H5N1 isolates. While, the amino acids associated with sensitivities to various antiviral drugs (Oseltamivir, Zanamivir, Amantadine) were found conserved for the Pakistani H5N1 isolates. Conspicuously, some important mutations observed at critical positions of antigenic sites (S141P, D155S, R162I & P181S) and at receptor binding pocket (A185T, R189K & S217P) of HA-1. A high sequence similarity between Pakistani HP H5N1 and LP H9N2 viruses was also observed. Avian like host specific markers with the exception of E627K in PB2, K356R in PA, V33I in NP, I28V in M2 and L107F in NS2 proteins were also observed. CONCLUSIONS: Various point mutations in different genes of H5 viruses from Pakistan were observed during its circulation in the field. The outbreaks started in Khyber Pakhtoon Khawa (North West) province in 2006 and spread to the Southern regions over a period of time. Though migratory birds may have a role for this continued endemicity of clade 2.2 H5N1 viruses during 2006-2008 in Pakistan, the possibility of their transmission through legal or illegal poultry trade across the borders cannot be ignored.

PIXE analysis of PM2.5 and PM(2.5-10) for air quality assessment of Islamabad, Pakistan: application of chemometrics for source identification.

A Gent sampler was used to collect 379 pairs of filters from Nilore, a suburban area of Islamabad city. The study was designed to assess the concentration variations of trace elements in fine and coarse particulate matter due to anthropogenic activities and naturally occurring events. Source identification was performed by applying MATLAB software for principal component analysis (PCA), and cluster analysis (CA). The average fine and coarse particulate masses during the study period were 15.1 ± 11.9 and 37.3 ± 28.0 µg/m(3) respectively which complies with the 24-h air quality limits set by the government of Pakistan. The application of PCA to PM(2.5) data suggests the PM contribution from sources such as soil, automobile exhaust and coal combustion, road dust and wearing of tyres, wood combustion, biomass burning and fertilizers and fungicides whereas for the PM(2.5-10) data shows signatures of suspended soil, automobile exhaust, road dust and wearing of tyres, wood and biomass burning, refuse incineration, Ni smelter, fertilizers and fungicides are obtained. Cluster analysis of PM(2.5) and PM(2.5-10) datasets reveals that there are mainly three contributory pollution sources and these are suspended soil particles, automobile related sources and wood and coal combustion.

Pollution level and health risk assessment of road dust from an expressway.

Thirteen road dust and four soil samples were analyzed using instrumental neutron activation analysis (INAA) to determine the elemental composition of road dust collected from Islamabad Expressway. The amounts of most of the elements determined were generally in the lower range of the global data with the exception of Ce, Nd, Sn and Zr, which were measured at higher concentrations in Islamabad. The magnitudes of Enrichment Factors (EFs) showed that the area around Islamabad Expressway is low to moderately polluted especially by elements such as Mg and Sb. Elemental health risk was assessed through dose calculations for carcinogenic and non-carcinogenic metals; and by the determination of LADD (lifetime average daily dose). The carcinogenic metals studied showed the following variation in their LADD values; Cr>Ni>Co>Cd>As. Dose calculations for non-cancerous and cancerous elements showed the data obtained to lie within the safe threshold of 10(-3)-10(-6) mg kg(-1) day(-1) for most elements. The Soil-based elements Al, Fe, K, Mg, Na and Ti were the exception and originated from the exposed soil around the highway. Moreover highest Hazard Index (HI) was found to be associated with the presence of Al, Cr, Pb and V in road dust showing soil, transport and industry to be the major sources of road dust.

H7 avian influenza virus vaccines protect chickens against challenge with antigenically diverse isolates.

Vaccination has been a critical tool in the control of some avian influenza viruses (AIV) and has been used routinely in Pakistan to help control sporadic outbreaks of highly pathogenic (HP) H7 AIV since 1995. During that time, several AIV isolates were utilized as inactivated vaccines with varying degrees of success. In order to evaluate which H7 AIV strains may serve as optimal vaccines for diverse H7 AIVs from Pakistan we conducted vaccination-challenge studies with five H7 vaccines against challenge with two HPAIVs: A/chicken/Murree/NARC-1/1995 H7N3 and A/chicken/Karachi/SPVC-4/2004 H7N3. To further characterize the isolates antigenic cartography was used to visually demonstrate the antigenic relationships among the isolates. All vaccines provided similar protection against mortality, morbidity and shedding of challenge virus from the respiratory tract. However, some minor (not statistically significant) differences were observed and correlated with antibody levels induced by the vaccine prior to challenge.

Sequence and phylogenetic analysis of H7N3 avian influenza viruses isolated from poultry in Pakistan 1995-2004.

BACKGROUND: Avian influenza virus (AIV) infections have caused heavy economic losses to the poultry industry in Pakistan as well as numerous other regions worldwide. The first introduction of H7N3 AIV to Pakistan occurred during 1995, since then H7N3, H9N2 and H5N1 AIVs have each been sporadically isolated. This report evaluates the genetic origin of the H7N3 viruses from Pakistan collected 1995-2004 and how they disseminated within the country. To accomplish this we produced whole genome sequences for 6 H7N3 viruses and data for the HA and NA genes of an additional 7 isolates. All available sequence from H7N3 AIV from Pakistan was included in the analysis. RESULTS: Phylogenetic analysis revealed that there were two introductions of H7 into Pakistan and one N3 introduction. Only one of the H7 introductions appears to have become established in poultry in Pakistan, while the other was isolated from two separate outbreaks 6 years apart. The data also shows that reassortment has occurred between H7N3 and H9N2 viruses in the field, likely during co-infection of poultry. Also, with the exception of these few reassortant isolates, all 8 genes in the predominant H7N3 virus lineage have evolved to be phylogenetically distinct. CONCLUSIONS: Although rigorous control measures have been implemented in commercial poultry in Pakistan, AIV is sporadically transmitted to poultry and among the different poultry industry compartments (broilers, broiler breeders, table egg layers). Since there is one primary H7 lineage which persists and that has reassorted with the H9N2 AIV in poultry, it suggests that there is a reservoir with some link commercial poultry. On a general level, this offers insight into the molecular ecology of AIV in poultry where the virus has persisted despite vaccination and biosecurity. This data also illustrates the importance of sustained surveillance for AIVs in poultry.

Evaluation of dietary status with respect to trace element intake from dry fruits consumed in Pakistan: a study using instrumental neutron activation analysis.

Instrumental neutron activation analysis has been utilized for the quantification of 24 major, minor and trace elements in dried apricots (Prunus armeniaca), dates (Phoenix dectylifera), figs (Ficus carica) and raisins (Vitis vinifera) using a low-power reactor as the neutron source. The weekly intakes of aluminium, arsenic, boron, calcium, chlorine, cobalt, chromium, iron, mercury, potassium, sodium, magnesium, manganese, molybdenum, antimony, selenium and zinc have been calculated and compared with the Recommended Dietary Allowance/Estimated Safe and Adequate Dietary Intake values through consumption of 100 g dried fruit. Fairly adequate levels of calcium, cobalt, iron, potassium and magnesium were determined in these dried fruits while the contribution of chlorine, chromium, molybdenum and sodium to the Recommended Dietary Allowance was found to be insignificant. Apricot is a nutritionally rich source of potassium, magnesium and cobalt, while chlorine and manganese contents were found to be highest in dates. Figs are an ample source of calcium, iron, magnesium and manganese, whereas raisins provide the highest intake of chromium, molybdenum and sodium. The toxic element contents of these dry fruits are considerably below the tolerance limits.

Urban air quality in the Asian region.

Over the past decade, member states of the Regional Co-operation Agreement (RCA), an intergovernmental agreement for the East Asia and Pacific region under the auspices of the IAEA with the assistance of international organizations and financial institutions such as the World Bank and the Asian Development Bank, have started to set in place policies and legislation for air pollution abatement. To support planning and evaluate the effectiveness of control programs, data are needed that characterizes urban air quality. The focus of this measurement program describe in this report is on size segregated particulate air pollution. Such airborne particulate matter can have a significant impact on human health and urban visibility. These data provide the input to receptor models that may permit the mitigation of these impacts by identification and quantitative apportionment of the particle sources. The aim of this report is to provide an overview of the measurements of concentrations and composition of particulate air pollution in two size fractions across the participating countries. For many of the large cities in this region, the measured particulate matter concentrations are greater than air quality standards or guidelines that have been adopted in developed countries.