Background: -During In vitro fertilization technology, embryo potential for
implantation and successful pregnancy is usually evaluated depending on
mostly morphological criteria. Embryo fragmentation is often used as the
main criterion for qualifying the human embryo grade and its implantation
potential. Fragments of human embryoblastomeres are apoptotic bodies and
or anuclear cytoplasmic pieces that release mitochondrial DNA (mtDNA)
into the embryo secretome. Cell freemtDNA content of human embryo
secretome can be used as a noninvasive tool to evaluate the fragmentation of
the embryo cells and thus its grade and implantation potential.
Aim: the aim of the present research was to evaluate the correlation between
Mitochondrial DNA content of embryo culture medium and human embryo
blastomeres fragmentation as an objective noninvasive criterion for embryo
quality.

Materials and Methods:-50 spent embryo culture mediawere collected
from17 Intracytoplasmic sperm injection (ICSI) cycles.After extraction and
purification of the free circulating DNA, mitochondrial DNA (mtDNA) was
profiled by specific semi quantitative method (Agarose gel
electrophoresis).Agarose gel electrophoresis revealed the amplification of the
mtDNA from many samples but with different concentrations (the intensity
of the bands were vary greatly) and some other samples failed to give a
positive amplification.

Results: - mtDNA was semi quantified in 50 spent media samples collected
from 17 female patients. In 12 samples (24%) mtDNA was undetectable. 6
samples showed mild mtDNA profile, 21 samples with moderate mtDNA
profile and 11 samples showed severe mtDNA profile. Insignificant
correlation was found between mtDNA profiling in D3 of embryonic
development and embryo fragmentation, but when correlating mtDNA
profile on D3 and embryo fragmentation and grade; highly significant
correlation was found with embryos of G2 or lower quality.

Conclusion: -Results of the work shown in the present paper are suggestive
of a significant correlation between the human embryo culture media
mtDNA content and the degree of embryo blastomeres fragmentation
especially in bad quality embryos; that may constitute a promising
noninvasive objective criterion for human embryo quality grading and for
selecting single embryo transfer policy.

The endemic highly pathogenic H5N1 avian influenza viruses (A/H5N1) of clade 2.2.1 in Egypt
compromise the poultry industry and pose a serious public health threat. In spite of vaccination, infections of
commercial poultry flocks have been frequently reported. The 2.2.1.2 viruses were isolated from vaccinated
commercial poultry and are postulated to be immune escape variants (IEV). In a trial to control the wide spread
of Highly pathogenic avian influenza (HPAI) H5N1 virus outbreaks among poultry flocks in Egypt this study
has been pursued to Evaluate the efficacy of possible available AI (H5N1, H5N2 and H5N3) vaccines against
challenging with the current recently isolated HPAI H5N1 (A/duck/Egypt/CLEVB-24_N00238/2015) field
strain this trail was planned to explain why some poultry farms are vaccinated against AI and suffered from
outbreaks of HPAI infection. in this study 5 commercially available inactivated avian influenza vaccines were
evaluated for their efficacy using both SPF and broiler chickens, SPF groups were vaccinated at 21st
day old
while broiler groups were vaccinated at 10th
day old to simulate field condition, the groups were serologically
monitored on a weekly basis post vaccination (PV) using HI test, then challenge test were conducted at 28th

PV for SPF groups and at 28th
day old for broiler groups using the recently isolated (A/duck/Egypt/CLEVB-
24_N00238/2015) field strain s challenge virus, oropharyngeal swab samples were collected for detection of
virus shedding titer by real time RT- PCR). The results of HI against the homologous antigens for each vaccine
showed positive seroconversion for the 5 H5 AI vaccines in both SPF and broiler chickens, Meanwhile only 3
H5 vaccines included positive seroconversion against the heterologous A/duck/Egypt/CLEVB-24_N00238/2015
HPAI antigen with mean HI titers of 5.9, 4.7 and 3.9 log2 in broiler chickens and 7.5, 5.1 and 4 log2 in SPF
chickens for poulvac H5N3, Nobilis H5N2 and Re-5 H5N1 AI vaccines respectively. While the two local Re-
H5N1 AI vaccines included HI titers < 3 (negative seroconversion) in both broiler and SPF chickens. These
mean HI values were reflected on the protection percentages which were 90%, 85%, 85%, 50% and 40% in
broiler chickens and 90%, 85%, 90%, 55% and 45%, in SPF chickens vaccinated with poulvac H5N3, Nobilis
H5N2, Re-H5N1, Mevac H5N1 and serovac H5N1 AI vaccines, respectively. Finally concerning the reduction
on the challenged HPAI H5N1 virus shedding, only the same 3 vaccines (poulvac H5N3, Nobilis H5N2 and Re
H5N1 – induced significant reduction in the titer of the shedding virus (more than 2 logs) compared to the non-
vaccinated challenged groups in both broiler and SPF chickens while the rest two did not provide significant
reduction in the titer of shedding virus compared to the non-challenged control group in both boiler and SPF
chickens.

Poultry production has experienced tremendous change in Egypt in the last three
decades. Small-scale family poultry production, otherwise termed household
poultry, was part of this transformation but to date no concise description has
been made of Egyptian household poultry. In this report, this is described using
surveys and reviews. Inputs and outputs of this production system were evaluated
and the pro
fi
tability of the household poultry was estimated. Household poultry
contribute immensely to food security in Egypt; providing income for individual
families. A mean
fl
ock size of 73 (mixed breeds) was determined and this yielded a
net annual pro
fi
t of 2287.67LE (US$397.34) per annum. The important household
poultry diseases are principally viral and bacterial. While Egyptian household
poultry are similar to others in Africa in terms of multi-species
fl
ocks, women-
driven projects, labour and marketing structures, it differs in input systems,
hatchery method, disease management, and other indices. Suggestions for
improvement of this sector of the poultry industry are offered.

Domestic ducks have been implicated in the
dissemination and evolution of H5N1 highly pathogenic
avian influenza (HPAI) viruses. In this study, two H5N1
HPAI viruses belonging to clade 2.2.1 isolated in Egypt in
2007 and 2008 were analyzed for their pathogenicity in
domestic Pekin ducks. Both viruses produced clinical signs
and mortality, but the 2008 virus was more virulent,
inducing early onset of neurological signs and killing all
ducks with a mean death time (MDT) of 4.1 days. The
2007 virus killed 3/8 ducks with a MDT of 7 days. Full-
genome sequencing and phylogenetic analysis were used to
examine differences in the virus genes that might explain
the differences observed in pathogenicity. The genomes
differed in 49 amino acids, with most of the differences found in the hemagglutinin protein. This increase in path-
ogenicity in ducks observed with certain H5N1 HPAI
viruses has implications for the control of the disease, since
vaccinated ducks infected with highly virulent strains shed
viruses for longer periods of time, perpetuating the virus in
the environment and increasing the possibility of trans-
mission to susceptible birds.

This study was carried out to investigate the efficacy of mono-specific antisera prepared in rabbits for the detection and
differential diagnosis of classic and variant strains of infectious bronchitis virus (IBV) in comparison with those prepared in
specific pathogen free (SPF) chickens in addition to their ability in detection of the degree of cross neutralization and antigenic
relationship between these strains. Two serological tests were used; indirect haemagglutination inhibition test (IHI) and serum
neutralization test (SNT). Also, two classic (H120, MA5) and two variant (4/91, CR88) IB antigens were used. IHI test gave
higher titers not only in case of homologous strains than heterologous but also higher in rabbits than chickens (H120 strain Vs
H120 antisera 12 log2 in rabbits and 9.6 log2 in chickens. The results of IHI were confirmed by using SNT that classic (H120) and
variant (4/91) strains gave high neutralizing index (NI) when using antisera for rabbit (0.5) while in chickens H120 and CR88
gave (3.8). So, rabbit is recommended animal for preparation of commercial monospecific hyperimmune sera which can be
successfully used in detection and differentiation of classic and variant strains of IBV and evaluation of antigenic relationship
between these strains.

Formaldehyde concentration is very crucial in the potency of inactivated vaccines and its adverse
effects. High level of formaldehyde will affect on potency of the vaccine by masking the B and T cell epitopes.
Determination of formaldehyde concentration can be measured by visual method (phenyl hydrazine) and by
spectrophotometric methods (phloroglucinol and ferric chloride methods). In the current study, the sensitivity
and specificity of these methods were investigated. The visual method is not accurate where it is depended on
naked eye in matching of a coloured complex product while the two spectrophotometric methods gave nearly
the same values and required inexpensive instrumentation(spectrophotometer) with simple operation. The ferric
chloride quantitative colorimetric method depends on complicated principle, easily used in routine analysis and
the used chromogenic agent is fairly expensive. In addition, this method is highly sensitive and accurate
compared to phloroglucinol method where it could determine the formaldehyde concentration of all inactivated
veterinary vaccines either bacterial or viral. Also, it could differ between the oil adjuvanted and the gel
adjuvantedinactivated vaccines during the operating process.

In the present study, several molecular techniques were used to analyze the two serovars (A and C) that used in the quality control assays of Avibacterium paragallinarum (A. paragallinarum) vaccines attained to our laboratory (Central Laboratory For Evaluation Of Veterinary Biologics CLEVB). Western blotting analysis clearly revealed a differences in bands intensity when reacted to antisera prepared against either serovar A or C especially at area of 40-55 KDa. On the other hand nucleotide sequence analysis could revealed three single nucleotide polymorphisms (SNPs) between serovar A and C at position of 17(T/C) , 46 ( G/A) and 178 ( T/C) and one area of deletion in serovar C at nucleotide position 94 – 102. Hence these findings represent a good molecular marker for conformity and differentiation between the two tested serovars.

In the present study, several molecular techniques were used to analyze the two serovars (A and C) that used in the quality control assays of Avibacterium paragallinarum (A. paragallinarum) vaccines attained to our laboratory (Central Laboratory For Evaluation Of Veterinary Biologics CLEVB). Western blotting analysis clearly revealed a differences in bands intensity when reacted to antisera prepared against either serovar A or C especially at area of 40-55 KDa. On the other hand nucleotide sequence analysis could revealed three single nucleotide polymorphisms (SNPs) between serovar A and C at position of 17(T/C) , 46 ( G/A) and 178 ( T/C) and one area of deletion in serovar C at nucleotide position 94 – 102. Hence these findings represent a good molecular marker for conformity and differentiation between the two tested serovars.

In a trial to control the wide spread of Highly pathogenic avian influenza (HPAI) H5N1 virus
outbreaks among poultry flocks in Egypt, many inactivated oil adjuvant AI virus vaccines were used. All
of these vaccines were either low pathogenic H5N2 AI viruses or genetically re-assorted H5N1 AI virus. In
the current study a recombinant fowl pox-avian influenza (AI) H5 vaccine (reFP-AIV-H5); expressing the
hemagglutinin of the A/turkey/Ireland/1378/83 (H5N8) AI isolate; was evaluated in comparison with the
genetically re-assorted inactivated H5N1strain A/Goose/Guandong/1/96 and inactivated H5N2
strainA/CK/Mexico/232/CPA/94 in SPF chickens. The potency of the 3 vaccines using HI test against
homologous and heterologous AI antigens were 5, 10.2 and 7.7 log2 HI unites, respectively. While, HI
titre against the heterlogous AI local antigen prepared from A/chicken/Egypt/12378 N3-CLEVB/2006/H5N1
strain were 2, 6.4 and 5 log2 HI unites, respectively and 0, 4.6 and 0 log , respectively against the heterologous 2
AI local antigen prepared from A/chicken/Egypt/9402 NAMRU3-CLEVB 213/2007/H5N1 strain. On the
other hand, the efficacy of the 3 vaccines in SPF chickens was studied. The protection percentages were
40, 90 and 80% against HPAI isolate 2006 and were 0, 31 and 19 against HPAI isolate 2007, respectively. AIV
shedding was detected and titrated in both vaccinated and control challenged birds. It was concluded that
the reFP-AIV-H5 vaccine is not suitable to be used to protect poultry flocks in Egypt against the circulating
AIV either 2006 or 2007 strains.