Houssam Shaib

Evidence of infection with H4 and H11 avian influenza viruses among Lebanese chicken growers.

Human infections with H5, H7, and H9 avian influenza viruses are well documented. Exposure to poultry is the most important risk factor for humans becoming infected with these viruses. Data on human infection with other low pathogenicity avian influenza viruses is sparse but suggests that such infections may occur. Lebanon is a Mediterranean country lying under two major migratory birds flyways and is home to many wild and domestic bird species. Previous reports from this country demonstrated that low pathogenicity avian influenza viruses are in circulation but highly pathogenic H5N1 viruses were not reported. In order to study the extent of human infection with avian influenza viruses in Lebanon, we carried out a seroprevalence cross-sectional study into which 200 poultry-exposed individuals and 50 non-exposed controls were enrolled. We obtained their sera and tested it for the presence of antibodies against avian influenza viruses types H4 through H16 and used a questionnaire to collect exposure data. Our microneutralization assay results suggested that backyard poultry growers may have been previously infected with H4 and H11 avian influenza viruses. We confirmed these results by using a horse red blood cells hemagglutination inhibition assay. Our data also showed that farmers with antibodies against each virus type clustered in a small geographic area suggesting that unrecognized outbreaks among birds may have led to these human infections. In conclusion, this study suggests that occupational exposure to chicken is a risk factor for infection with avian influenza especially among backyard growers and that H4 and H11 influenza viruses may possess the ability to cross the species barrier to infect humans.

Modulation by essential oil of vaccine response and production improvement in chicken challenged with velogenic Newcastle disease virus

To evaluate the impact of Eucalyptus and peppermint essential oils on immune modulation and production of broiler chicken challenged with a molecularly characterized velogenic NewCastle disease virus (vNDV).

Evaluation of a Salmonella Enteritidis vaccine and related ELISA for respective induction and assessment of acquired immunity to the vaccine and/or Echinacea purpurea in Awassi Ewes.

The aim of this study was to evaluate an experimental Salmonella Enteritidis (SE) bacterin and an indirect ELISA system to assess quantitatively the acquired immunity in Awassi ewes to the vaccine and/or Echinacea purpurea (EP) dried roots. Four treatments of the ewes were included in the experimental design, with 6 ewes/treatment. The first treatment (T1) had the controls that were non-vaccinated and non-treated with EP. The T2 ewes were only treated with EP. The T3 and T4 ewes were vaccinated at D1 (initiation of trial) and D10, while the T4 ewes were additionally administered the EP dried roots. Blood was collected from the jugular vein of all ewes at D1, D10, D21 and D45. The construction of the vaccine and the ELISA are detailed within the manuscript. The ELISA was able to detect quantitatively the significant acquired primary and secondary immunity to the vaccine in T3 and T4 ewes, compared to their low level of background immunities at initiation of the experiment (p<0.05). In addition, the ELISA detected the absence of seroconversion at all blood sampling times (p>0.05) in T1 control ewes, and in the T2 ewes that were given only the (EP) (p>0.05). Moreover, the ELISA was able to uncover the significant seroconversion of secondary immune response in T4 ewes at D21 compared to that at D10 (p<0.05), and the absence of significant seroconversion of secondary response in T3 ewes. This is the first work in literature that reports the need to supplement the vaccination by the experimental SE bacterin with daily oral intake of 250mg of EP-dried roots, effective the first vaccination day and up to 21 days, for obtaining a statistically significant seroconversion.

Comparison of phenotypic and virulence genes characteristics in human and chicken isolates of Proteus mirabilis

Abstract The objective of this work is to compare the phenotypic and virulence genes characteristics in human and chicken isolates of Proteus mirabilis. The bacterial examination of 50 livers of individual broilers, marketed by four major outlets, revealed a high recovery of P. mirabilis (66%), and a low recovery frequency of Salmonella spp. (4%), Serratia odorifera (2%), Citrobacter brakii (2%), and Providencia stuartii (2%). The phenotypic biochemical characterization of the recovered 33 chicken isolates of P. mirabilis were compared to 30 human isolates (23 urinary and six respiratory isolates). The comparison revealed significant differences in the presence of gelatinase enzyme (100% presence in chicken isolates versus 91·3 and 83·3% presence in human urinary and respiratory isolates, respectively, P<0·05). The H2S production occurred in 100% of chicken isolates versus 95·6 and 66·7% presence in human urinary and respiratory isolates, respectively, P<0·05). The other 17 biochemical characteristics did not differ significantly among the three groups of isolates (P>0·05). Two virulence genes, the mrpA and FliL, were having a typical 100% presence in randomly selected isolates of P. mirabilis recovered from chicken livers (N = 10) versus isolates recovered from urinary (N = 5) and respiratory specimens of humans (N = 5) (P>0·05). The average percentage similarity of mrpA gene nucleotide sequence of poultry isolates to human urinary and respiratory isolates was 93·2 and 97·5-%, respectively. The high similarity in phenotypic characteristics, associated with typical frequency of presence of two virulence genes, and high similarity in sequences of mrpA gene among poultry versus human P. mirabilis isolates justifies future investigations targeting the evaluation of adaptable pathogenicity of avian Proteus mirabilis isolates to mammalian hosts.

A Mini Review of qRT-rtPCR Technology Application in Uncovering the Mechanism of Food Allergy and in the Search for Novel Interventions

This mini review targets the inclusion of recent selected citations, between the year 2006 and 2012, that implement the qRT-rtPCR technology in their experimental designs, targeting the uncovering of the mechanism of food allergy. In addition, this same technology was implemented in specific experimental designs, aiming at finding novel nutritional, herbal medicine, and tolerance interventions against food allergy. The approach of using qRT-rtPCR technology helped in studying the dynamics of transcription of cytokines and chemokines in intestinal dendritic cells of the experimental animals during the allergic reaction to food. The suppression of transcription of specific cytokines or chemokines by nutritional, herbal medicine, and tolerance interventions was instrumental in the search for finding novel remedies for this health condition, that was traditionally managed by avoidance of offending foods in the diet.

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.

Evaluation of Antibiotics to Control Mycoplasma gallisepticum in Broiler Breeder Chickens

This study aims at the evaluation of the efficacy of Pulmotil and Denagard in comparison to the generic tylosin against Mycoplasma gallisepticum (MG) infection in broiler breeders. A total of 600 twenty four-week old broiler breeder pullets along with their 60 roosters of the Ross 308 strain, were equally subdivided into four treatments of 150 birds each with 3 replicates per treatment (50 pullets and 5 roosters/pen). The MG-free birds of Group 1, the Control, were kept in a separate house and left with no MG-challenge or drug administration while those of groups 2, 3, and 4 were raised in another house and previously challenged with MG at 2 and 24 weeks of age respectively. Every 4 weeks and for 3 consecutive days, birds of group 3 were administered Pulmotil (3–21 Weeks) and Denagard (25–44 weeks), whereas those of Group 4 were treated throughout the trial with generic Tylosin. Production performance parameters were not significantly different among the differently treated birds. At 30 weeks of age, Pulmotil-Denagard treatment significantly reduced the tracheal MG counts to a similar level to that of the controls and restored the fertility of the MG challenged birds at 39 weeks of age. At 35 weeks of age, Pulmotil-Denagard treatment significantly reduced the frequency of day-old progeny with positive airsac lesions. It also reduced MG colonization in the airsacs of the day-old offsprings of the 35 and 39 week old breeders. As the Pulmotil-Denagard treatment significantly reduced the MG colonization of bird tissues, it reduced MG sera titers at 12, 20 and 30 weeks of age, and increased titers against IB, IBD and ND at later stages. In conclusion, the use of Pulmotil/Denagard program is highly recommended in MG-infected breeder farms, and protects against any potential MG endemic infection.

Dietary Hemicell® Improved Reproductive Performance of Broiler Breeders under Commercial Settings

The objective of this work is to study the effect of dietary Hemicell®, a fermentation product that contains high amount of β-mannanase, on performance and health parameters of broiler breeders grown under commercial settings. Twenty six thousand broiler breeders of Ross 308 strain with an average egg production of 35% and 2600 roosters aged 26.5 weeks were used in this study and were subdivided equally into six individual environmentally controlled houses with 4333 hens per house and 10% roosters. Birds in three houses were fed regular diet provided by the Ross Company while those in the remaining three other houses were fed rations containing 250 g of Hemicell® per ton of finished feed for a period of 8.6 months. Hemicell® in breeder diets reduced feed cost and numerically improved number of total and hatching eggs per hen housed. It also increased hatchability by 0.32% during the 8.6 months trial period. Although Hemicell® improved yolk colour, it had no effect on other egg quality parameters such as egg weight and shell thickness. Regarding health parameters, Hemicell® reduced the incidence of pododermatitis in hens and increased the serum titer for NDV towards the end of the trial. Along with the little saving in feed cost, Hemicell® resulted in a gain of around 1.20 US dollar per hen housed. These results indicate that Hemicell® inclusion in the diets of broiler breeders is beneficial to parent flock growers and makes their operations more profitable.

Characterization of a canarypox virus from an outbreak among canaries (Serinus canaria domesticus) in Lebanon

ABSTRACT A canarypox outbreak resulted in the death of about 50% of canaries (Serinus canaria domesticus) in several breeding farms in Lebanon. Infected birds showed thickened eye lid and skin scab-like lesions at the beak, foot and caudal regions and died 5–6 days post disease symptoms onset. Out of seven sick birds that were autopsied for gross pathology evaluation, one bird demonstrated turbid airsacs with absence of any lesions in the trachea, oesophagus, liver and lung. Histopathological examination showed hyperplasia, heterophils infiltration and Bollinger body formation in the skin of five out of the seven autopsied canaries. Hyperplasia and infiltration of heterophils were also observed in the airsacs of one bird. PCR analysis of specimens taken from the skin and feet, targeting the fpv167 gene of the canarypox virus, was positive for all of the analyzed canaries. PCR analysis also revealed that two birds had concurrent infection with Mycoplasma gallisepticum. Sequencing and alignment of the amplified fpv167 gene of the canarypox showed 100% similarity with the Iranian canary pox isolate IR/H913/14. Smuggling of pet birds through the borders should be strictly controlled and biosecurity measures must be adequately applied to control the circulation of the two identified pathogens.

Standardization of a Protocol for Quantitative Evaluation of Anti-AerosolizedInfluenza Virus Activity by Vapors of a Chemically-Characterized Essential OilBlend

The aim of this research is to standardize the conditions of a constructed impinger, enabling to evaluate quantitatively the anti-aerosolized H9N2 avian influenza virus (AIV) activity by vapors of a chemically-characterized essential oil blend. The standardization resulted in 100% recovery of the aerosolized H9N2 virus when the impinger’s conditions were set at aerosolized viral particles count of 1.2 × 106/c.c. of Tryptose Phosphate Broth, temperature of 35°C, average micelle diameter of 44.3 μm, negative pressure of 6 mbar, air-suspension time of H9N2 virus of 1.5 min, collection chamber and its transport medium volumes of 250 cc and 25 cc, respectively. The adoption of the above standardized conditions, with an inclusion of vaporized essential oil (EO) at 1.0 × 10-4 μl EO/μl volume of the pulverization chamber, and contact times of 0.5-1.5 min with the H9N2 virus, resulted in 84.6% reduction in viral titer at 1.5 min contact time, compared to the control virus, deprived from contact with vaporized EO (P<0.05). This new finding will help in future investigations related to application of safe essential oils in reduction of air-suspended influenza virus in closed systems harboring domestic animals and human populations.