The stabilisation of clayey soil by using sawdust and sawdust ash.

Soil stabilisation by waste materials has been recently employed to enhance soil engineering properties. The purpose of this study is to compare the impact of utilising sawdust in its raw form versus sawdust ash as a soil stabiliser. This is to determine if sawdust (SD) can be considered as a substitute for sawdust ash (SDA) in order to reduce incineration and air pollution. To fulfil this aim, the Atterberg limits, modified Proctor test, and Direct Shear test were performed on both stabilised and non-stabilised mixtures of clayey soil. The soil was treated with 2%, 5%, 8%, 12%, 15%, and 20% by soil dry weight of both SD and SDA. The findings show that the use of SD and SDA leads to a reduction in the plasticity index and the maximum dry unit weight of the soil while increasing its optimum moisture content. The bearing capacity of the soil was greatest at 5% for both SD and SDA, with SD exhibiting a greater enhancement (31.89%) than SDA. Therefore, it is recommended to utilise SD instead of SDA for soil stabilisation due to its superior effectiveness and less harmful environmental impact.

Whole-genome sequencing of the clinical isolate of Legionella pneumophila ALAW1 from the West Bank allows high-resolution typing and determination of pathogenicity mechanisms.

Background: Legionella pneumophila is water-based bacterium causing Legionnaires' disease (LD). We describe the first documented case of nosocomial LD caused by L. pneumophila sequence type (ST) 461 and serogroup 6. The etiology of LD was confirmed by culturing the bronchoalveolar lavage sample retrieving L. pneumophila strain ALAW1. A 7-days treatment of the LD patient with Azithromycin and Levofloxacin allowed complete recovery. Methods: In details, we sequenced the whole genome of the L. pneumophila ALAW1 using Illumina HiSeq platform. The sequence of ALAW1 was aligned with the genome sequence from the closely related reference strain Alcoy 2300/99 and a whole-genome phylogeny based on single nucleotide polymorphisms (SNPs) was created using Parsnp software. Also, the TYGS web-server was used in order to compare the genome with type strain. Results: An analysis of the population structure by SNP and TYGS comparison clustered ALAW1 with the reference genome Alcoy 2300/99. Blastp analysis of the type IV secretion Dot/Icm system genes showed that these genes were highly conserved with (≤25%) structural differences at the protein level. Conclusions: Overall, this study provides insights into detailed genome structure and demonstrated the value of whole-genome sequencing as the ultimate typing tool for Legionella.

Cytotoxicity, Intracellular Replication, and Contact-Dependent Pore Formation of Genotyped Environmental Legionella pneumophila Isolates from Hospital Water Systems in the West Bank, Palestine.

Legionella pneumophila is the causative agent of Legionnaires' disease. Due to the hot climate and intermittent water supply, the West Bank, Palestine, can be considered a high-risk area for this often fatal atypical pneumonia. L. pneumophila occurs in biofilms of natural and man-made freshwater environments, where it infects and replicates intracellularly within protozoa. To correlate the genetic diversity of the bacteria in the environment with their virulence properties for protozoan and mammalian host cells, 60 genotyped isolates from hospital water systems in the West Bank were analyzed. The L. pneumophila isolates were previously genotyped by high resolution Multi Locus Variable Number of Tandem Repeat Analysis (MLVA-8(12)) and sorted according to their relationship in clonal complexes (VACC). Strains of relevant genotypes and VACCs were compared according to their capacity to infect Acanthamoeba castellanii and THP-1 macrophages, and to mediate pore-forming cytotoxicity in sheep red blood cells (sRBCs). Based on a previous detailed analysis of the biogeographic distribution and abundance of the MLVA-8(12)-genotypes, the focus of the study was on the most abundant L. pneumophila- genotypes Gt4(17), Gt6 (18) and Gt10(93) and the four relevant clonal complexes [VACC1, VACC2, VACC5 and VACC11]. The highly abundant genotypes Gt4(17) and Gt6(18) are affiliated with VACC1 and sequence type (ST)1 (comprising L. pneumophila str. Paris), and displayed seroroup (Sg)1. Isolates of these two genotypes exhibited significantly higher virulence potentials compared to other genotypes and clonal complexes in the West Bank. Endemic for the West Bank was the clonal complex VACC11 (affiliated with ST461) represented by three relevant genotypes that all displayed Sg6. These genotypes unique for the West Bank showed a lower infectivity and cytotoxicity compared to all other clonal complexes and their affiliated genotypes. Interestingly, the L. pneumophila serotypes ST1 and ST461 were previously identified by in situ-sequence based typing (SBT) as main causative agents of Legionnaires' disease (LD) in the West Bank at a comparable level. Overall, this study demonstrates the site-specific regional diversity of L. pneumophila genotypes in the West Bank and suggests that a combination of MLVA, cellular infection assays and hierarchical agglomerative cluster analysis allows an improved genotype-based risk assessment.

MiR-16-1-3p and miR-16-2-3p possess strong tumor suppressive and antimetastatic properties in osteosarcoma.

Osteosarcoma (OS) is an aggressive malignancy affecting mostly children and adolescents. MicroRNAs (miRNAs) play important roles in OS development and progression. Here we found that miR-16-1-3p and miR-16-2-3p "passenger" strands, as well as the "lead" miR-16-5p strand, are frequently downregulated and possess strong tumor suppressive functions in human OS. Furthermore, we report different although strongly overlapping functions for miR-16-1-3p and miR-16-2-3p in OS cells. Ectopic expression of these miRNAs affected primary tumor growth, metastasis seeding and chemoresistance and invasiveness of human OS cells. Loss-of-function experiments verified tumor suppressive functions of these miRNAs at endogenous levels of expression. Using RNA immunoprecipitation (RIP) assays, we identify direct targets of miR-16-1-3p and miR-16-2-3p in OS cells. Moreover, validation experiments identified FGFR2 as a direct target for miR-16-1-3p and miR-16-2-3p. Overall, our findings underscore the importance of passenger strand miRNAs, at least some, in osteosarcomagenesis.

Comparison of in situ sequence type analysis of Legionella pneumophila in respiratory tract secretions and environmental samples of a hospital in East Jerusalem.

Legionella pneumophila genotyping is important for epidemiological investigation of nosocomial and community-acquired outbreaks of legionellosis. The prevalence of legionellosis in pneumonia patients in the West Bank was monitored for the first time, and the sequence types (STs) from respiratory samples were compared with STs of environmental samples from different wards of the hospital. Sputum (n = 121) and bronchoalveolar lavage (BAL) (n = 74) specimens were cultured for L. pneumophila; genomic DNA was tested by 16S rRNA polymerase chain reaction (PCR) amplification. Nested PCR sequence-based typing (NPSBT) was implemented on DNA of the respiratory and environmental PCR-positive samples. Only one respiratory specimen was positive for L. pneumophila by culture. BAL gave a higher percentage of L. pneumophila-positive samples, 35% (26/74) than sputum, 15% (18/121) by PCR. NPSBT revealed the following STs: ST 1 (29%, 7/24), ST 461 (21%, 5/24), ST 1037 (4%, 1/24) from respiratory samples, STs from environmental samples: ST 1 (28.5%, 4/14), ST 187 (21.4%, 3/14) and ST 2070, ST 461, ST 1482 (7.1%, 1/14) each. This study emphasises the advantage of PCR over culture for the detection of L. pneumophila in countries where antibiotics are indiscriminately used prior to hospital admission. ST 1 was the predominant ST in both respiratory and environmental samples.

Oseltamivir activity against avian influenza H9N2 strain with different point mutations in their neuraminidase.

The present study has two aims: to optimize the antiviral activity of oseltamivir in chicken embryos against an avian influenza-H9N2 strain (P0) and to apply the optimized protocol for studying the drug susceptibility of 4 H9N2 mutants (M1, M2, M3, and M4). As for the first aim, oseltamivir antiviral activity was monitored upon its delivery into 9-day-old chicken embryo at a concentration of 0.27 mg/100 µl, against 7 doses of the P0 strain, ranging between 1.2 x 10(-5) and 2.0 Hemagglutination (HA) units. Oseltamivir showed its highest efficacy in reduction of viral propagation (95% reduction in HA titer) (P 〈0.05), when the inoculum level contained a minimum HA units of 1.2 x 10(-5). For the second aim of this study, the application of the 1.2 x 10-(5) HA units of the virus in inocula for the evaluation of oseltamivir-antiviral effect against the 4 H9N2 mutants revealed an emergence of a resistant mutant (M1), associated with 2 adjacent point mutations in its neuraminidase (N) amino acid (aa) sequence at positions 46 and 47. The other 3 mutants maintained a variable sensitivity to oseltamivir, resulting in the following reduction in HA titers: M2 (82.9%), M3 (61.5%), and M4 (100.0%). How the point mutations of the neuraminidase sequences affected the susceptibility of H9N2 virus to oseltamivir is still to be determined and deserve further investigations.

Optimization of Saanen sperm genes amplification: evaluation of standardized protocols in genetically uncharacterized rural goats reared under a subtropical environment.

The purpose of this research is to optimize quantitatively the amplification of specific sperm genes in reference genomically characterized Saanen goat and to evaluate the standardized protocols applicability on sperms of uncharacterized genome of rural goats reared under subtropical environment for inclusion in future selection programs. The optimization of the protocols in Saanen sperms included three production genes (growth hormone (GH) exons 2, 3, and 4, αS1-casein (CSN1S1), and α-lactalbumin) and two health genes (MHC class II DRB and prion (PrP)). The optimization was based on varying the primers concentrations and the inclusion of a PCR cosolvent (Triton X). The impact of the studied variables on statistically significant increase in the yield of amplicons was noticed in four out of five (80%) optimized protocols, namely in those related to GH, CSN1S1, α-lactalbumin, and PrP genes (P < 0.05). There was no significant difference in the yield of amplicons related to MHC class II DRB gene, regardless of the variables used (P > 0.05). The applicability of the optimized protocols of Saanen sperm genes on amplification of uncharacterized rural goat sperms revealed a 100% success in tested individuals for amplification of GH, CSN1S1, α-lactalbumin, and MHC class II DRB genes and a 75% success for the PrP gene. The significant success in applicability of the Saanen quantitatively optimized protocols to other uncharacterized genome of rural goats allows for their inclusion in future selection, targeting the sustainability of this farming system in a subtropical environment and the improvement of the farmers livelihood.

Impact of competitive non-protective antigens in a booster killed vaccine on seroconversions to protective antigens of Newcastle disease virus in chickens.

This study was designed to examine the impact of competitive non-protective antigens in a bivalent killed vaccine of Newcastle disease virus and infectious bronchitis (IB) virus on seroconversions against protective fusion protein of Newcastle disease (ND) virus (NDV), in free-range layers primed by live ND-clone 30 and IB-H120 vaccines. The experimental design included two free-range layer farms in which sera of randomly chosen layers were collected on two occasions from each of the two farms namely: at the time of administration of the killed booster vaccine (23 weeks of age) and three weeks later. The Western immunoblotting technique was used to react the individual pooled sera collected at different times from each farm with antigens used in priming, namely those of the ND-clone 30 virus and the IB-H120 virus. The optical density bands formed by reactions were compared statistically between seroconverted antibodies at 23 weeks with those of layers aged 26 weeks. The killed booster vaccine offered a significant seroconversion on both farms to the non-protective L-protein (248.0 kDa) of NDV and on one of the two farms to the non-protective NDV-matrix protein (40.0 kDa) (p<0.05). However, seroconversion to the protective fusion protein of NDV (60 kDa) failed on both farms (p<0.05). In addition, on one farm, a statistical significance was revealed by the killed booster vaccine seroconversion to non-protective IBV-nucleocapsid protein (510 kDa) and, on the other farm, to another non-protective IBV-glycoprotein (28.0 kDa) (p<0.05). The impact of competitive seroconversions to non-protective antigens and seroconversion failures to low molecular weights of NDV protective fusion protein is discussed.

Impact of synergism of calcium phosphomycin/tylosin tartarate supplements on the performance of broilers in the Lebanon.

The authors evaluate the impact of a synergistic preparation (SP) of supplements (a combination of calcium phosphomycin and tylosine tartarate) on the performance of broilers with a history of carcass condemnation at slaughter. The experiment included 120-day old broilers (Ross 308), divided equally into two treatment groups, with three replicates per treatment and 20 birds per replicate. The two groups included controls that did not receive SP and those that were treated with SP. The SP group received treatment at three intervals (at 1-5 days of age: 160 mg/kg body weight; at 21-25 days of age: 80 mg/kg; and at 29-33 days of age: 80 mg/kg body weight). The administration of SP at a low level improved performance in SP birds compared to controls and also resulted in the lowest cumulative mortality (1.67% vs 6.67%, respectively), the lowest feed conversion of 1.91 between 1 and 43 days of age and the highest live body weight (2,544.75 g vs 2,390.18 g). The administration of SP at a low level improved performance and reduced the frequency of specific gross lesions at market age (tracheitis, lung congestion, breast blisters and bursal congestion).

Standardisation of a new model of H9N2/Escherichia coli challenge in broilers in the Lebanon.

Primary infection by low pathogenic avian influenza (LPAI) predisposes for secondary infection by Escherichia coli in poultry, leading to significant economic losses. Future research in control of this ailment requires the establishment of a successful controlled challenge by avian influenza virus (AIV)/E. coli. Six groups of broilers (6 birds/group) were included for the standardisation of the controlled challenge by AIV/E. coli. Birds in groups 1, 2, 3, 4 and 5 received an intra-tracheal challenge of 0.5 ml of two haemagglutinating units of H9N2 virus at 20 days of age. At the age of 23 days, birds in group 1 received an intra-thoracic (right air sac)-E. coli challenge equivalent to 1.6 x 10 colony-forming units (cfu)/0.5 ml/bird, while birds in groups 2, 3, 4 and 5 received E. coli by the same route and in the following respective decreasing order of viable cells: 1.6 x 10(6), 1.6 x 10(5), 1.6 x 10(4) and 1.6 x 10(3); cfu. Birds in control group 6 were deprived of H9N2 and E. coli challenge. Results showed significant early mortality in group 1 that was challenged with the highest number of E. coli, in comparison to groups 2-6 (p<0.05); however, the average weight at 28 days of age was similar in surviving birds of groups 2-6 (p>0.05). The frequencies of four signs at 2 days and at 5 days post E. coli challenge (conjunctivitis, diarrhoea, ocular exudates and rales) in the surviving birds of groups 2-5 were most often higher than those observed in control group 6 (p<0.05). These four signs and five gross lesions (abdominal airsacculitis, left thoracic airsacculitis, pericarditis, right thoracic airsacculitis and tracheitis) had a decreasing pattern of frequency related to a decrease in the E. coli count used in the challenge.

Comparison of the level of organic contaminants in bile and muscle of Mugil spp. following a major oil spill in the eastern Mediterranean Sea.

The total polyaromatic hydrocarbons (TPAH) and the total polychlorinated biphenyls (TPCB) that originate from oil spills in sea and ocean waters are toxic to fish and their offspring. The authors compare the levels of organic contaminants (TPAH and TPCB) recovered from the bile versus the dorsal muscles of 120 individual Mugil spp. that were harvested from six sites in the eastern Mediterranean following a significant heavy oil spill. Results showed an insignificant difference between the mean of means of TPAH and TPCB (six means of individual Mugil spp. from six respective sites) in bile versus dorsal muscle. In addition, the correlation equation between the level of bile TPAH and the level of bile carcinogenic polyaromatic hydrocarbons (cPAH) was established. This data suggests the possibility of substituting the analysis of organic contaminants in muscles by using the liquid bile of Mugil spp., thus eliminating the time-consuming steps of lyophilisation and homogenisation of muscle. In addition, the bile cPAH could be predicted from the bile TPAH by a regression relationship.