Efficiency of natural oils as alternative adjuvants to mineral oils in inactivated avian influenza vaccine formulation.

1. The effectiveness of inactivated vaccines depends on selecting the suitable adjuvant for vaccine formulation. The potency of vaccines with low antigen content can be improved with the appropriate adjuvant. This could allow production of more doses and lower the production cost.2. This study evaluated the efficiency of vaccines prepared using oil extracted from natural sources including argan oil, almond oil, sesame seed oil, pumpkin oil, cactus oil and black seed oil as alternative adjuvants for improving the protection capacity of inactivated influenza virus vaccine as compared to commonly used mineral oils.3. Each vaccine formulation was evaluated for stability, safety and immunogenicity in chickens, as well as for reducing the viral shedding after challenge infection.4. The cactus, sesame and pumpkin seed oil-based vaccines were found to be potent and successfully induced the production of humoral immunity in vaccinated chickens.

Adenovirus type D and type E infection in broiler chickens: the effect on CD4 and CD8 T cell response, cytokines expression and their immunopathology.

1. A total of 150-day-old chicks were divided into three groups of 50 birds (G1-G3); G1 and G2 were orally inoculated at 1-day old with 0.5 ml of 107 TCID50/ml FAdV-D serotype 2 (MT386509.1) and FAdV-E serotype 8a (MW847902), respectively, and G3 was blank control group.2. Cell-mediated immune response was evaluated by detection of CD4, CD8 T lymphocytes and the mRNA expression of IL6 and IL8 in the chicken spleen using q-PCR. Additionally, immunopathology was performed at 3, 5 and 7 day post infection (dpi) and weekly until the end of the experiment.3. Results revealed that transcription of inflammatory cytokines (IL6, IL8) was up regulated in the spleen of FAdV type D and type E infected chickens at various time points relative to the control group. A marked decrease in the number of CD4 and CD8 T lymphocytes at 5 and 7 dpi in G1 of chickens infected with FAdV type D. Whereas, in chickens infected with FAdV type E, the CD4 and CD8 T lymphocytes were markedly decreased at 7 dpi.4. In contrast, there were no significant differences in humoral immune responses against NDV vaccine in (G1 and G2) at different intervals post-vaccination compared to the control group. The histopathology of the bursa, thymus, and spleen in the infected groups showed lymphocytolysis with severe reticular cells hyperplasia and lymphoid depletion.5. In conclusion, fowl adenovirus types D and E have an immunosuppressive effect in broilers which may be considered one of the main causes of the continuous co-infections with other viruses reported in the field during the last 10 years.

Efficacy of live attenuated vaccine derived from the Streptococcus agalactiae on the immune responses of Oreochromis niloticus.

Streptococcus agalactiae species have been recognized as the main pathogen causing high mortality in fish leading to significant worldwide economical losses to the aquaculture industries. Vaccine development has become a priority in combating multidrug resistance in bacteria; however, there is a lack of commercial live attenuated vaccine (LAV) against S. agalactiae in Malaysia. The aim of this study is to compare two methods using attenuated bacteria as live vaccine and to evaluate the efficacy of selected LAV on the immune responses and resistance of Oreochromis niloticus (tilapia) against S. agalactiae. The LAV derived from S. agalactiae had been weakened using the chemical agent Acriflavine dye (LAV1), whereas the second vaccine was weakened using serial passages of bacteria on broth media (LAV2). Initial immunization was carried out only on day one, given twice-in the morning and evening, for the 42 day period. Serum samples were collected to determine the systemic antibody (IgM) responses and lysozymal (LSZ) activity using ELISA. On day 43 after immunization, the fish were injected intraperitoneally (i.p) with 0.1 mL of S. agalactiae at LD50 = 1.5 × 105 (CFU)/fish. Fish were monitored daily for 10 days. Clinical signs, mortality and the relative percent of survival (RPS) were recorded. Trial 1 results showed a significant increased (P < 0.05) in serum IgM titers and LSZ activity as compared to LAV2 and the control group (unvaccinated fish). The efficacy of LAV1 was proven effective as determined by the RPS values, LAV1 at 81.58% as compared to LAV2 at 65.79%. Trial 2 of LAV1 and control group were further determined by administering primary and booster doses revealed a RPS value for LAV1 of 82.05%, with the significant enhancement on the immune responses of tilapia as compared to control group. In conclusion, LAV revealed to elevate antibody IgM levels, LSZ activity and provide long-term protection when added to feed. LAV is a low-cost vaccine shown to rapidly increase the immune response of fish and increase survival rates of fish against S. agalactiae infection.

Mutations in foot and mouth disease virus types A and O isolated from vaccinated animals.

Foot and mouth disease (FMD) is responsible for serious economic losses in Egypt. Although vaccination is practised as the main control strategy, failure of vaccination has been reported in many cases, which can be due to a number of factors. Selection of FMD antigenic variants under the immune pressure of partially immunised hosts has been previously recorded. This study was designed to isolate and characterise foot and mouth disease viruses (FMDVs) circulating in Egyptian vaccinated animals. Serotype O, A and Southern African Territories (SAT) 2 FMDVs were detected in different Egyptian governorates during 2015, 2016 and 2017. The successful isolation of 15 FMDVs of the three serotypes is reported in this paper. Phylogenetic analysis based on the viral protein (VP) 1 gene showed that all serotype O isolates had East Africa (EA)-3 topotypes. There was variation in 15-17 amino acids between the serotype O isolates of 2015 and those of 2016 and 2017. The serotype A isolates belonged to the A-Iran-05 lineage, with the exception of one isolate of 2016 which clustered with the African strains of G-IV. Serotype SAT2 FMDV was detected in two samples of 2017 and both were of lineage Alx-12 of topotype VII. The virus neutralisation test using sera raised against vaccine strains confirmed the serotyping of the isolates and determined the antigenic relatedness between the isolates and the currently used vaccine strains. A decrease in the neutralising antibody titre of some serotype O and A isolates could be attributed to mutation in critical amino acids in the neutralising antigenic sites. Hence, this work supports previous studies describing the significance of amino acid substitutions within the antigenic sites of the virus in antibody neutralisation and immune escape.

La fièvre aphteuse est à l'origine d'importantes pertes économiques en Égypte. Si la vaccination constitue la principale stratégie de lutte contre cette maladie, de nombreux échecs vaccinaux ont été rapportés, dus à différents facteurs. Il a été constaté par le passé que la pression immunitaire exercée par des hôtes partiellement immunisés contre la fièvre aphteuse entraînait une sélection de variants antigéniques du virus. La présente étude avait donc pour objet d'isoler et de caractériser les virus de la fièvre aphteuse présents en Égypte chez les animaux vaccinés. Les sérotypes O, A et Southern African Territories (SAT) 2 du virus de la fièvre aphteuse ont été détectés dans plusieurs gouvernorats égyptiens en 2015, 2016 et 2017. Les auteurs font état de 15 isolements réussis de souches virales appartenant à chacun des trois sérotypes. Il ressort de l'analyse phylogénétique basée sur le gène de la protéine virale 1 (PV1) que tous les isolats de sérotype O correspondaient au topotype East Africa (EA) 3. Une variation de 15 à 17 acides aminés a été observée entre les virus de sérotype O isolés en 2015 et ceux du même sérotype isolés en 2016 et en 2017. Les isolats de sérotype A appartenaient tous à la lignée A-Iran-05, à l'exception d'un isolat de 2016 qui était proche des souches africaines du lignage G-IV. Le sérotype SAT-2 du virus de la fièvre aphteuse a été détecté dans deux échantillons prélevés en 2017. Les deux souches isolées appartenaient à la lignée Alx-12 du topotype VII. La neutralisation virale utilisant des antisérums produits contre les souches vaccinales a permis de confirmer le sérotypage des souches isolées et de déterminer le degré de similitude entre les isolats et les souches vaccinales utilisées actuellement. La diminution du titre d'anticorps neutralisants dirigés contre certains isolats des sérotypes O et A est probablement imputable à une mutation d'acides aminés déterminants au sein des sites de neutralisation des antigènes. Ce travail corrobore les résultats d'études antérieures qui avaient révélé que les substitutions d'acides aminés au sein des sites antigéniques du virus peuvent avoir un rôle dans la neutralisation d'anticorps et dans l'échappement du virus au système immunitaire.

La fiebre aftosa es causante de graves pérdidas económicas en Egipto. Aunque la vacunación viene siendo la principal estrategia de lucha, se han descrito numerosos casos en los que ha resultado ineficaz, hecho que puede deberse a varios factores. Anteriormente ya se había observado que la presión inmunitaria de hospedadores parcialmente inmunizados conduce a la selección de determinadas variantes antigénicas de la fiebre aftosa. Los autores exponen un estudio encaminado a aislar y caracterizar los virus de la fiebre aftosa circulantes en los animales egipcios vacunados. En los años 2015, 2016 y 2017 se detectaron en diferentes provincias del país los serotipos víricos O, A y SAT (Southern African Territories) 2. Los autores dan cuenta del aislamiento de 15 virus de la fiebre aftosa pertenecientes a uno u otro de estos tres serotipos. El análisis filogenético basado en el gen de la proteína vírica (VP) 1 demostró que todos los virus del serotipo O aislados correspondían al topotipo EA (East Africa)-3, con diferencias localizadas en los aminoácidos 15 a 17 entre los virus aislados en 2015 y los de 2016 y 2017. Los virus del serotipo A pertenecían al linaje A-Iran-05, con la excepción de uno de los de 2016, que formaba un conglomerado con las cepas africanas del linaje G-IV. El serotipo SAT2, por su parte, estaba presente en dos muestras de 2017, pertenecientes ambas al linaje Alx-12 del topotipo VII. Empleando la prueba de neutralización vírica con sueros sensibilizados contra cepas vacunales se confirmó el serotipo de los virus aislados y se determinó el grado de parentesco antigénico entre esos virus y las cepas vacunales utilizadas actualmente. El decremento observado en el título de anticuerpos neutralizantes frente a algunos de esos virus de los serotipos O y A podría explicarse por la mutación de aminoácidos fundamentales de los sitios antigénicos neutralizantes. Este trabajo, por lo tanto, viene a corroborar anteriores estudios que señalaban la gran influencia de la sustitución de aminoácidos en los sitios antigénicos del virus en los procesos de neutralización de anticuerpos y escape inmunitario.

Histological and immunohistochemical investigations of ovarian interstitial glands during non-breeding season in camels (Camelus dromedarius).

The aim of this was to investigate the histology and immunohistochemistry of interstitial glands during non-breeding season in camel ovaries. A total of 21 mature, non-pregnant and apparently healthy camels aged between 8 and 12 years were slaughtered. The ovaries were removed within 15 min, cleaned from adipose tissue, weighted and examined grossly. The histological preparation was made, and then, the blocks were cut at 3-5 microns thickness and stained by H&E for histological examinations. Moreover, some sections were stained with Sudan Black for lipid detection. Immunohistochemical analysis of paraffin-embedded ovarian tissues was performed to detect the localization of S-100, vimentin, progesterone receptors (PR) and oestrogen receptors (ER). Immunoreactive signals were detected using UltraVision Detection System. The results revealed that the interstitial glands were located in the cortical region and they were arranged in various arrangements either single, in couple or in groups rich in lipid droplet. All interstitial gland arrangements were enclosed by connective tissue capsules containing fibroblasts and collagenous fibres separated them from the surrounding ovarian structures. Both interstitial glands and their surrounding CT were penetrated by several blood vessels. There was a strong immunoreactive signal for S-100 in the nuclei of interstitial cells, and no signals were detected either in cells of the interstitial glands or their connective tissue with PR. We could conclude that the interstitial gland is distinct in ovary of camel and further studies are needed to elucidate its rule in steroid synthesis.

Pathogenicity of the Egyptian A/H5N1 avian influenza viruses in chickens.

Long-term circulation of highly pathogenic avian influenza H5N1 viruses of clade 2.2.1 in Egyptian poultry since February 2006 resulted in the evolution of two distinct clades: 2.2.1.1 represents antigenic-drift variants isolated from vaccinated poultry and 2.2.1.2 that caused the newest upsurge in birds and humans in 2014/2015. In the present study, nine isolates were collected from chickens, ducks and turkeys representing the commercial and backyard sectors during the period 2009-2015. The subtyping was confirmed by hemagglutination inhibition (HI) test, RT-qPCR and sequence analysis. The Mean Death Time (MDT) and Intravenous Pathogenicity Index (IVPI) for all isolates were determined. Sequence analysis of the HA gene sequences of these viruses revealed that two viruses belonged to clade 2.2.1.1 and the rest were clade 2.2.1.2. Antigenic characterisation of the viruses supported the results of the phylogenetic analysis. The MDT of the isolates ranged from 18 to 72 h and the IVPI values ranged from 2.3 to 2.9; viruses of the 2.2.1.1 clade were less virulent than those of the 2.2.1.2 clade. In addition, clade-specific polymorphism in the HA cleavage site was observed. These findings indicate the high and variable pathogenicity of H5N1 viruses of different clades and host-origin in Egypt. The upsurge of outbreaks in poultry in 2014/2015 was probably not due to a shift in virulence from earlier viruses.

Protective efficacy of a prime-boost protocol using H5-DNA plasmid as prime and inactivated H5N2 vaccine as the booster against the Egyptian avian influenza challenge virus.

In this study, a recombinant DNA plasmid was constructed, encoding for HA1 of a selected Egyptian H5N1 virus (isolated during the 2012 outbreaks). In the immunization and challenge experiments, SPF chickens received 1 or 2 doses of H5-DNA plasmid prime, and boosted with the inactivated H5N2 vaccine. Haemagglutination inhibition (HI) titers, protection levels, and the magnitude of virus shedding were compared to that of the chickens that received either DNA plasmid or inactivated H5N2 vaccine alone. H5N1 virus A/chicken/Egypt/128s/2012 (H5N1) highly pathogenic avian influenza (HPAI) clade 2.2.1/C was used for the challenge. Chickens immunized with 1 or 2 doses of H5-DNA vaccine failed to overcome the challenge with 0% and 10% protection, respectively. Quantitative real-time reverse transcription-PCR revealed virus shedding of 2.2 x 104 PCR copies/ml 3 days post challenge (dpc) in the only surviving bird from the group that received 2 doses of plasmid. However, chickens immunized with 1 or 2 doses of H5-DNA plasmid as prime and inactivated H5N2 vaccine as booster, showed 80% protection after challenge, with a viral shedding of 1.2 x 104 PCR copies/ml (1 dose) and 1.6 x 104 PCR copies/ml (2 doses) 3 dpc. The surviving birds in both groups did not shed the virus at 5 and 7 dpc. In H5N2-vaccinated chickens, protection levels were 70% with relatively high virus shedding (1.8 x 104 PCR copies/ml) 3 dpc. HI titers were protective to the surviving chickens. This study reports the efficacy of H5-DNA plasmid to augment reduction in viral shedding and to provide better protection when applied in a prime-boost program with the inactivated AI vaccine.

Assessment of serum adiponectin in Egyptian patients with HCV-related cirrhosis and hepatocellular carcinoma.

BACKGROUND AND AIM: Hepatitis C-associated insulin resistance is involved in the development of various complications including hepatocarcinogenesis. Low plasma levels of adiponectin contribute to the pathogenesis of insulin resistance and type II diabetes mellitus. The aim of this study was to determine the value of serum adiponectin in Egyptian patients with hepatitis C-related cirrhosis and hepatocellular carcinoma. PATIENTS AND METHODS: 90 Egyptian patients with hepatitis C-related liver cirrhosis were enrolled in this study. Patients were divided into two groups as follows: group I: 61 patients with hepatitis C-related cirrhosis and hepatocellular carcinoma, group II: 29 patients with hepatitis C-related cirrhosis (hepatocellular carcinoma was excluded in these patients at the time of recruitment in the study). Serum adiponectin level was measured and correlated with all other studied parameters. RESULTS: Serum adiponectin was significantly lower in patients with hepatocellular carcinoma and it had significant negative correlations with both the overall tumor size and the number of tumor foci. Highly significant negative correlations were found between adiponectin and all markers of insulin resistance in both groups. At a cut-off value ≤5.4 µg/ml, adiponectin had a sensitivity of 60.7%, a specificity of 93.1%, a positive predictive value of 94.9%, and a negative predictive value of 52.9% for detection of hepatocellular carcinoma (with an overall accuracy of 77.6%). CONCLUSION: An independent association exists between serum adiponectin and hepatocellular carcinoma in Egyptian patients with hepatitis C-related cirrhosis. Therapy to increase circulating adiponectin concentration might represent a novel strategy to prevent hepatitis C-related hepatic complications.

Effect of long-term testosterone propionate or human chorionic gonadotrophin administration on reproductive glands in adult male rabbits.

The study was aimed to investigate the effect of testosterone propionate (TP) or human chorionic gonadotrophin (hCG) treatment on reproductive glands in sexually mature male rabbits. A total 36 adult male rabbits were randomly distributed to six equal groups. The first control group (CON), the second treated with low-dose TP (TPL), the third treated with high-dose TP (TPH), the fourth treated with low-dose hCG (CGL), the fifth treated with medium-dose hCG (CGM) and sixth treated with high-dose hCG (CGH). At the 16th post-treatment week, the animals were sacrificed, and the testes and accessory sex glands dissected, weighted and stored at -20 °C until assay. Testosterone propionate treatment in both doses resulted in reduction (P < 0.01) in testicular weight and increase (P < 0.01) in weight of vesicular gland, paraprostate and proprostate glands. High-dose TP increased the weight of prostate and bulbouretheral gland (BUG). Testosterone propionate increased total androgen (P < 0.01) with Testosterone (T) predominating in serum, dihydrotestosterone (DHT) predominating in testes and most accessory sex glands. High dose of hCG increased the weight of proprostate and paraprostate glands. Androgen level in serum, testes and accessory sex glands increased (P < 0.01) after hCG treatment.

Terminal 2q deletion and partial trisomy chromosome 15q: a clinical and cytogenetic study.

We report on a 5 years old female patient with a karyotype 46, XX, add (2), t(2;15) (q37;q22) associated with dysmorphic facial features, digital deformities, heart defect (mild mitral regurge) and severe mental retardation. This is the third reported case worldwide on the terminal 2q deletion and trisomy of chromosome 15q syndrome. The findings in this case and our literature review, delineates the pattern of malformations secondary to trisomy of 15q and deletion of 2q.

Acid-base parameters and steroid concentrations in pre-ovulatory follicles and plasma of lactating dairy cows with spontaneous and synchronized oestrus or follicular cyst.

The study aimed to compare the acid-base balance and steroid concentrations between follicular fluids (FF) of pre-ovulatory follicles derived from a spontaneous oestrus (SO), synchronized or induced oestrus (IO) and follicular cysts (CYS) and between FF and blood in dairy cows. Forty-two dairy cows were included in this study. The animals were allocated to three groups: SO (n = 23); IO (n = 11) using GnRH at day 0 and day 9 and PGF2 α at day 7; and animals with CYS (n = 10). The follicular fluids (FF) were aspirated from the cyst/pre-ovulatory follicles (∅ ≥ 15 mm) after SO and after second GnRH dose in IO by transvaginal ultrasound-guided ovum pick-up technique. Blood samples (BL) were collected in heparinized vacutainer tubes. The oxygen tension (pO2) in FF of IO was higher (p < 0.05) than in SO and CYS groups. There were negative correlations (p < 0.001, r = -0.89) between FF and blood pO2. The carbon dioxide tension (pCO2) and lactate level in FF of CYS group were higher (p < 0.05) than in SO and IO groups. There were negative correlations (p < 0.01, r = -0.73) between blood and FF pO2. Oestradiol-17ß concentration in pre-ovulatory follicles and plasma of the SO group was higher (p < 0.001) than in IO and CYS groups. Progesterone concentration in pre-ovulatory follicles and plasma of the SO and IO groups was lower (p < 0.01) than in CYS group. Plasma androstenedione concentration in SO and IO groups was higher (p < 0.05) than in CYS group. In conclusion, acid-base parameters, E2 and P4 levels in the follicular fluid of both IO and CYS groups were deviated greatly from the physiological level (disturbances of intrafollicular/intracystic environment), which may affect the quality of both the oocyte and the granulosa cells.

The emerging role of lncRNA MEG3 and MEG3 rs7158663 in hepatocellular carcinoma.

OBJECTIVE: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy in Egypt. Genetic and environmental factors play a role in its development. This study explored the association between the long non-coding RNA (lncRNA) MEG3 rs7158663 polymorphism, MEG3 expression, and the risk of HCC and other clinicopathologic characteristics in an Egyptian population. PATIENTS AND METHODS: This case-control study included 114 patients with HCC and 110 healthy controls. TaqMan Real-time PCR was used to analyze lncRNA MEG3 rs7158663. Serum MEG3 expression levels were measured using RT-PCR. RESULTS: The AA, GA+AA, and A alleles were associated with increased risk for HCC (adjusted odds ratio (OR) 11.84%, 95% CI 4.07-34.45, p < 0.0001; adjusted OR 3.18, 95% CI 1.79-5.67, p < 0.0001; and adjusted OR 2.87, 95% CI 1.91-4.34, p < 0.0001, respectively). The mutant genotype and allele were linked to an increased risk in male patients and patients ≥ 50 years old. MEG3 serum expression level was downregulated in HCC patients. The rs7158663 G > A polymorphism and downregulated MEG3 were significantly associated with larger tumor size and advanced disease stage. CONCLUSIONS: MEG3 rs7158663 single nucleotide polymorphisms and downregulated lncRNA MEG3 were associated with HCC risk and may represent diagnostic and bad prognostic factors for HCC patients.

Association of miR-146a rs57095329 with Behçet's disease and its complications.

BACKGROUND: Behçet's disease is a chronic relapsing and remitting autoimmune multisystem inflammatory disease characterised by oral aphthae, genital ulcers, skin lesions, gastrointestinal involvement, arthritis, vascular lesions and neurological manifestations. We hypothesised a link between rs57095329 of miR-146a and Behçet's disease, with further links with common clinical features. METHODS: We tested our hypothesis in 130 Behçet's disease patients and 131 age and sex-matched healthy controls. Behcet's disease current activity index (BDCAI) was used to assess patients' disease activity status. MiR-146a (rs57095329) was genotyped in all participants using RT-PCR and results in patients analysed according to clinical features. RESULTS: The frequency of the GG and AG genotypes in rs57095329 were strongly associated with Behçet's disease (adjusted OR 8.05, 95% CI 3.63-17.82; P < 0.001 and OR 2.26, 95% CI 1.27-4.04; P = 0.006, respectively), and in dominant (GG+AG > AA) and recessive (GG > AA+AG) models (both P < 0.001). Additionally, G allele distribution was significantly greater in Behçet's disease compared with controls (OR 2.85, 95% CI 1.98-4.11, P < 0.001). The AA genotype and A allele were linked to oral ulcers, the GG genotype and G allele to neurological disease, and the GG genotype and G allele to ocular disease (all P < 0.01). There were no links with genital ulceration, skin lesions, vascular disease or the result of the pathergy test. CONCLUSION: The miR-146a (rs57095329) is associated with Behçet's disease and certain genotypes and alleles with oral ulcers, and with ocular and neurological manifestations.

Impact of miR-155 (rs767649 A>T) and miR-146a (rs57095329 A>G) polymorphisms in System Lupus Erythematosus susceptibility in an Egyptian cohort.

OBJECTIVE: Systemic Lupus Erythematosus (SLE) is an autoimmune inflammatory disease. miR-155 and miR-146a were expressed in many autoimmune diseases such as rheumatoid arthritis. The aim of this study was to examine miR-155 rs767649 and miR-146a rs57095329 polymorphisms in SLE susceptibility in an Egyptian cohort and to investigate the correlation between them and clinical data and disease activity. PATIENTS AND METHODS: The two SNPs were analyzed in 120 patients with SLE and 100 healthy controls using RT-PCR. RESULTS: The TT genotype and T allele of miR-155 rs767649 were associated with a significant increase in the risk of SLE, particularly in females. On the other hand, miR-146a (rs57095329) polymorphism was not associated with SLE risk. The AT/TT genotypes of miR-155 rs767649 showed higher distributions among patients with higher SLEDAI and nephritis. CONCLUSIONS: This study had demonstrated for the first time the association between miR-155 rs767649 and the risk of development of SLE in an Egyptian cohort, mostly in females.

Genetic resistance of eight native Egyptian chicken breeds having chicken B-cell marker 6 gene post-challenge with field strain of Marek's disease-induced tumor virus.

AIM: The aim of this work was to detect chicken B-cell marker 6 (ChB6) gene in some native breeds in Egypt and find the relationship between founded genes in these different breeds to determine the resistance of native Egyptian breeds of chicken to Marek's disease (MD). MATERIALS AND METHODS: A total of 14 different chicken breeds (30 each) including ten native breeds in addition to SPF Lohmann, High Line, Bovans, and Roodiland were used. Blood samples were collected for the detection of (ChB6) by polymerase chain reaction (PCR) assay and sequenced to determine the presence or absence of ChB6 gene. Experimental infection was done using local field isolated MD virus (MDV) of 11 (1 day old) unvaccinated chick breeds having no maternal antibodies against MDV. Ten breeds of them carry ChB6 gene, eight breeds were native, and the rest two breeds were SPF Lohmann and High Line in addition to a group of ChB6 gene-lacking breed (Bovans) were infected. Spleen samples were collected from all infected breeds at 20th, 25th, 30th, 35th, and 40th weeks post-infection and tested by PCR assay for the detection of MDV. Furthermore, at 40th week post-infection, tumorized spleen sample of Bovans breed was collected and prepared for examination by transmission electron microscope (TEM) to confirm the presence of MDV. RESULTS: Our results revealed the positivity of 10 out of 14 breeds (Gimmizah, Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) to the presence of ChB6 gene and resistance to MDV infection, while the Bovans, Mandarah, Inshas and Roodiland breeds lack the ChB6 gene and are susceptible to MDV infection. The collected spleen samples revealed negative for the presence of challenged MDV by PCR in 10 breeds (Gimmizah, Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) and positive for Bovans breed. TEM is used to confirm MDV infection in Bovans group which demonstrated tumors. CONCLUSION: The study confirms the relationship between the presence of ChB6 gene in our native breeds and the absence of tumors.

Reverse spillover of avian viral vaccine strains from domesticated poultry to wild birds.

Transmission of viruses from the commercial poultry to wild birds is an emerging paradigm of livestock-wildlife interface. Here, we report the identification and isolation of vaccine strains of avian paramyxovirus serotype 1 (APMV1) and avian coronaviruses (ACoV) from different wild bird species across eight Egyptian governorates between January 2014 and December 2015. Surveillance of avian respiratory viruses in free-ranging wild birds (n=297) identified three species that harboured or excreted APMV1 and ACoVs. Genetic characterization and phylogenetic analysis of recovered viruses revealed a close association with the most widely utilized vaccine strains in the country. These results highlight the potential spillover of vaccine-viruses probably due to extensive use of live-attenuated vaccines in the commercial poultry, and close interaction between domesticated and wild bird populations. Further exploring the full spectrum of vaccine-derived viral vaccine strains in wild birds might help to assess the emergence of future wild-birds origin viruses.

Introduction and enzootic of A/H5N1 in Egypt: Virus evolution, pathogenicity and vaccine efficacy ten years on.

It is almost a decade since the highly pathogenic H5N1 avian influenza virus (A/H5N1) of clade 2.2.1 was introduced to Egypt in 2005, most likely, via wild birds; marking the longest endemic status of influenza viruses in poultry outside Asia. The endemic A/H5N1 in Egypt still compromises the poultry industry, poses serious hazards to public health and threatens to become potentially pandemic. The control strategies adopted for A/H5N1 in Egyptian poultry using diverse vaccines in commercialized poultry neither eliminated the virus nor did they decrease its evolutionary rate. Several virus clades have evolved, a few of them disappeared and others prevailed. Disparate evolutionary traits in both birds and humans were manifested by accumulation of clade-specific mutations across viral genomes driven by a variety of selection pressures. Viruses in vaccinated poultry populations displayed higher mutation rates at the immunogenic epitopes, promoting viral escape and reducing vaccine efficiency. On the other hand, viruses isolated from humans displayed changes in the receptor binding domain, which increased the viral affinity to bind to human-type glycan receptors. Moreover, viral pathogenicity exhibited several patterns in different hosts. This review aims to provide an overview of the viral evolution, pathogenicity and vaccine efficacy of A/H5N1 in Egypt during the last ten years.

[Perineal hypospadia in a red Holstein-calf. A case report]. / Perineale Hypospadie bei einem rotbunten Deutsche-Holstein-Kalb. Ein Fallbericht.

Hypospadia is a congenital deformation of the urethra. Several surgical correction methods have been described, but there are also cases when an operation is not necessarily required. In this report, the case of a bull calf which did not need an invasive therapy will be presented.

[Incidence of puerperal diseases during the first 10 days after foaling in the mare]. / Häufigkeit puerperaler Erkrankungen während der ersten 10 Tage post partum bei Stuten.

OBJECTIVE: The aim of this study was to evaluate the frequency of puerperal diseases in breeding mares in the first 10 days after birth by analysing patient data. MATERIAL AND METHODS: In a university clinic patient data of 308 breeding mares with puerperal disorders which presented within the first 10 days postpartum were evaluated over a period of 10 years. A distinction was made between diseases which were able to be diagnosed at the first examination and diseases which developed during the patient's stay in the clinic. RESULTS: A total of 21 diseases were diagnosed, with a retained placenta, lochiometra and injuries to the perineum being the most common. Many mares displayed more than one disease. Mares with a retained placenta most commonly also presented with perineal ruptures, followed by animals who also had lochiometra. Mares suffering from lochiometra commonly presented together with a retained placenta and injuries as a result of birth. Some of the mares developed further diseases. In mares with a retained placenta, this was most commonly lochiometra, followed by puerperal laminitis and thrombophlebitis. CONCLUSION AND CLINICAL RELEVANCE: The data collection shows that several diseases could relatively frequently be diagnosed in mares with puerperal disorders. Therefore, a higher percentage of further diseases must be assumed for mares which have a puerperal disease.

Effects of propylene glycol on the metabolic status and milk production of dairy buffaloes.

OBJECTIVES: The study was designed to investigate the effects of drenching with propylene glycol (PG) on body condition, serum metabolites and milk production during the transition period of dairy buffaloes. MATERIAL AND METHODS: Animals were randomly allocated to a control group (n=5) and a PG group of 10 buffaloes that were drenched with 500 ml of propylene glycol once daily from 10 (9±3) days prepartum until 2 weeks postpartum. Ultrasound measurements of backfat thickness (BFT) were performed weekly, while blood samples were taken at -4, -2, 2, 4, 6, and 8 weeks from parturition for estimation of hematological and biochemical metabolites. RESULTS: At -4, -3, and -2 weeks from calving, BFT did not differ between the two groups, but decreased after calving and was higher for the control group than the PG group at weeks -1 and 1. Hematological analysis revealed insignificant changes between the two groups. Serum concentrations of non-esterified fatty acids (NEFA), ß-hydroxybutyric acid (BHBA) and glucose did not differ between the two groups before parturition. At 2 and 4 weeks from parturition, NEFA was higher for the control group than the PG group. Serum concentrations of BHBA were higher at 2, 4, 6, and 8 weeks in control animals than in treated buffaloes. In contrast, the glucose level was significantly increased in PG group when compared to the control group at week 2 postpartum (p<0.05). Serum concentrations of total cholesterol, triglycerides, total proteins, albumin, and globulins did not differ significantly between the two groups (p>0.05). Serum enzyme activities of aspartate aminotransferase and γ-glutamyl transferase were significantly higher in the control than in the PG group. In treated buffaloes significantly (p<0.05) higher average 60-day milk yields were recorded (8.4±0.22 vs. 10.7±0.40 kg/day). Milk composition did not differ between the two groups. CONCLUSION AND CLINICAL RELEVANCE: Drenching of dairy buffaloes with propylene glycol may reduce the risk of ketosis, improve the metabolic status, and increase the milk yield.