2020, Scienceline Publication  
JWPR  
Poultry Research  
J. World Poult. Res. 10(2S): 247-249, June 14, 2020  
Journal of World’s  
Research Paper, PII: S2322455X2000030-10  
License: CC BY 4.0  
DOI: https://dx.doi.org/10.36380/jwpr.2020.30  
Detection of Bacterial Contamination in Imported Live Poultry  
Vaccines to Egypt in 2018  
Mohamed Morsi Elkamshishi1*, Haitham H. Ibrahim2 and Hanan M. Ibrahim3  
1Department of Animal Hygiene and Zoonoses, Faculty of Veterinary Medicine, Matrouh University, Egypt  
2Department of Avian and Rabbit Medicine, Faculty of Veterinary Medicine, Aswan University, Egypt  
3Head of researcher Central Laboratory for Evaluation of Veterinary Biologics (CLEVB) El-Seka El-Baida, Abbasia, Cairo, Egypt  
*Corresponding author’s Email: elkamshishymohamed@yahoo.com; ORCID: 0000-0002-2145-0663  
Received: 19 Feb. 2020  
Accepted: 29 Mar. 2020  
ABSTRACT  
The vaccine is one of the most important biological products used in the poultry industry, thus it must be safe, potent,  
and effective. This work presents the results of a large-scale diagnostic survey performed in Egypt to study hygienic  
epidemiology and how vaccination may affect the viral circulation in the field. This study aimed to detect bacterial  
contamination in live poultry vaccines imported to Egypt during 2018. In this study, 285 consignments poultry  
vaccines, including 114 consignments live vaccine, 103 consignments recombinant vaccines, and 68 consignments  
killed vaccines (imported through Cairo airport during 2018) were examined for bacterial contamination. The  
vaccines were imported from USA, Italy, France, Spain, Mexico, and China. Bacterial contamination with  
Salmonella species was detected using the VITEK 2 system in two samples (1.8%) (IB+HB1 vaccine imported from  
Italy and ILT vaccine imported from USA).  
Key words: Bacterial contamination, Egypt, Poultry, Vaccine  
INTRODUCTION  
even disappeared after the discontinuance of homologous  
vaccination (Franzo et al., 2014 and 2016). Live vaccine  
strains may spread to unvaccinated flocks, regain  
virulence because recombine with other circulating strains,  
possibly resulting in the emergence of new strains  
(Matthijs et al., 2008; Jackwood and Lee, 2017; Moreno  
et al., 2017).  
Hence, the aim of this work was to detect bacterial  
contamination in live poultry vaccines imported to Egypt  
during 2018.  
Egypt has a large sector of poultry, more than 50 thousand  
commercial producers and poultry farming systems (Ali  
et al., 2013).  
Egyptian requirements for imported animals and  
poultry by-products are governed by the epidemiological  
situation of the country and by the law of Agriculture No.  
53, 1966, and Ministerial Decree No. 47, 1967 concerning  
veterinary quarantine, and its modifications.  
Vaccines are widely used in the prevention and  
reduction of incidence as well as control of endemic  
poultry and animal diseases. For a vaccine to be useful in  
the maintenance of animal and poultry health, it must be  
pure, safe, potent, and effective (OIE, 2014).  
In Egypt, the Central Laboratory for Evaluation of  
Veterinary Biologics (CLEVB) is the authorized  
governmental laboratory for the evaluation and  
certification of veterinary vaccines and biological products  
prior to their release into the markets (Ministerial Decree  
No. 2978/2003). Despite essential to proper control of the  
disease, the massive use of live vaccines has some  
drawbacks. The impact of vaccination protocols on  
circulating strains proved remarkable and some lineages  
MATERIAL AND METHODS  
Vaccine samples  
As shown in Table 1, about 114 consignments of  
live poultry vaccines entered Egypt during 2018 each  
consignment represented by 5 vials.  
Table 1. Types and numbers of imported poultry vaccines  
to Egypt  
Recombinant  
vaccines  
Killed vaccines  
Live vaccines  
68 consignments  
103 consignments  
114 consignments  
To cite this paper: Elkamshishi MM, Ibrahim HH and Ibrahim HM (2020). Detection of Bacterial Contamination in Imported Live Poultry Vaccines to Egypt in 2018. J. World  
Poult. Res., 10 (2S): 247-249. DOI: https://dx.doi.org/10.36380/jwpr.2020.30  
247  
Elkamshishi et al., 2020  
The media used for detection of bacterial  
contamination  
RESULTS  
During 2018 Egypt imported 285 consignments poultry  
vaccines, including 114 consignments, live vaccine, 103  
consignments recombinant vaccines, and 68 consignments  
killed vaccines through Cairo airport. As shown in table 2,  
the vaccines imported from different countries as USA,  
Italy, France, Spain, Mexico, and China. All imported live  
vaccines were examined for bacterial contamination, two  
samples one is IB+HB1vaccine imported from Italy and  
the other was ILT vaccine imported from USA are  
detected bacterial contamination (by ratio 1.8%), these  
contaminated bacteria were identified as Salmonella  
species using VITEK 2 system.  
Broth media  
Tryptose broth (Non-selective enrichment) produced  
by Oxoid Company, batch No.147644/318 prepared and  
sterilized according to the manufacturer’s instruction.  
Selective isolated media  
Two selective isolated medium (MacConkey of  
batch No.2301758 code CMO115, produced by Oxoid  
company, and Salmonella-Shigella agar of batch  
NO.2235930 produced by Oxoid company are prepared  
and sterilized according to the manufacturer’s instruction.  
Xylose Lysine Deoxycholate Agar (XLD): Batch  
No.2283742 produced by Oxoid Company, prepared and  
sterilized according to the manufacturer’s instruction.  
Table 2. The import companies of poultry vaccines in  
Egypt  
Procedure  
The procedures are applied according to OIE (2016)  
1. Sample preparation:  
Import company  
Country of origin  
M.Ghannam  
USA, Italy, France, Spain, Mexico  
● If the vaccine sample contains 1000 doses or  
more so the vaccine was rehydrated at a rate of 30 ml sterile  
distal water/1000 dose.  
Intervet  
USA  
USA  
Axis Agency Services LTD.co.  
Zoetis Egypt  
USA  
● In case the vaccine sample in liquid phase the test  
portion will be 5 ml or one-half of the container contents,  
whichever is the lesser.  
Biopharma trading  
China  
International free trading  
company  
USA, Italy, France, Mexico  
2. Inoculation and incubation:  
●
Tryptose broth (Non-selective enrichment) 100  
ml is inoculated with 5 ml of the rehydrated sample or 50  
ml or one-half of the container contents, whichever is the  
lesser.  
98.2  
100  
● The inoculated broths are incubated for 18-24  
hours at 35-370 C.  
● Two selective isolated medium (MacConkey and  
Salmonella-Shigella agar) are inoculated from the  
nonselective broth (Tryptose broth) then incubated for 18-  
24 hours at 35-370 C and examined.  
50  
1.8  
0
negative samples  
positive samples  
● If on growth of typical Salmonella is noticed, the  
agar plates should be incubated an additional 18-24 hours at  
35-370 C and re-examined.  
● If colonies typical of Salmonella are observed,  
further subculture on to the Xylose Lysine Deoxycholate  
agar (XLD) is incubated at 35-370 C.  
Figure 1. Prevalence of bacterial contamination in  
imported live poultry vaccines to Egypt during 2018  
DISCUSSION  
3. Interpretation:  
● Interpretation depends on the presence or absence  
of the typical colony of Salmonella.  
● No growth (Not detected) means satisfactory  
results and Salmonella growth (detected) means  
unsatisfactory results.  
4. The identification:  
The bacterial contamination was identified as  
Salmonella species using the VITEK 2 system.  
Vaccine is still the most important tool for the control and  
prevention of poultry diseases. Their use in the poultry  
industry avoids or minimizes the clinical disease in the  
farms and increase production. Sterility testing is a process  
that confirms the presence or absence of any viable  
contaminating microorganisms in biological products  
(Lee, 1990).  
248  
J. World Poult. Res., 10(2S): 247-249, 2020  
use of IBV 793B vaccine needs reassessment after its withdrawal  
led to the genotype's disappearance. Vaccine, 32:6765–6767. DOI:  
10.1637/11557-121216-ResNote.1  
In this study, all live poultry viral vaccines enter  
Egypt during 2018 were examined for bacterial  
contamination, only two consignments out of 114 (1.8%)  
were refused due to contamination with Salmonella. Most  
researchers have carried out their investigation on the  
vaccine samples for the detection of viral contamination  
either by complete virus, antigens, or nucleic acids  
(Kamboh et al., 2009c). Kamboh et al. (2009a) recorded  
the 7.5% bacterial contamination in livestock vaccines and  
Kamboh et al. (2007) isolated 10.71% extraneous  
contaminants in local livestock vaccines used in Sindh  
province of Pakistan.  
Franzo G, Tucciarone CM, Blanco A, Nofrarías M, Biarnés M,  
Cortey M, Majó N, Catelli  
E and Cecchinato M (2016).  
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1952.1998  
Jackwood MW and Lee DH (2017). Different evolutionary trajectories of  
vaccine-controlled and non-controlled avian infectious bronchitis  
viruses in commercial poultry. PLoS One, 12(5):e0176709. DOI:  
10.1371/journal.pone.0176709  
The results obtained in the present study are in  
agreement with that obtained by Kamboh et al. (2009b)  
who found that lower bacterial contamination (2.08%) in  
poultry vaccines.  
Kojima A, Takahashi T, Kijima M, Ogikubo Y, Nishimura M, Nishimura  
S, Harasawa R and Tamura Y (1997). Detection of Mycoplasma in  
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In this study, the isolated bacterial contaminants  
were identified as Salmonella species using VITEK 2  
system which is a rapid system for identification of the  
gram-negative rods (including both members of the family  
Enterobacteriaceae and non-enteric bacilli) this system  
has been evaluated for identification of gram-negative  
bacilli by Guido et al. (1998). Samad (2001) reported the  
contamination of Bacillus megaterium, B. cereus, B.  
mycoids and B. subtilis in the local anthrax livespore  
vaccines. Whereas, Kojima et al. (1997) reported the  
contamination of avian Mycoplasma DNA in the avian  
live virus vaccines. Kamboh et al. (2009b) found E. coli in  
the vaccine of hydropericardium syndrome. Landman et  
al. (2000) found contamination of Marek’s disease vaccine  
by Enterococcus faecalis.  
Kamboh AA, Rind R, Shah AH, Gandahi JA and Rajput N (2007).  
Bacteriological study on local and imported livestock vaccines used  
in Sindh, Pakistan Journal of Agricultural Sciences, 3: 125–128.  
http://www.fspublishers.org  
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(2009a). Biochemical properties of bacterial contaminants isolated  
from livestock vaccines. Pakistan Journal of Nutrition, 8: 578–58.  
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Kamboh AA, Gandahi JA, Shah AH, Zhu WY, Malhi M,  
Rind R, and Babar ME (2009c). An evaluation for  
bacterial contamination of vaccines for buffaloes used  
in Pakistan. Pakistan Journal of Zoology, 9: 639-643.  
https://www.researchgate.net/publication/209836381.  
Kamboh AA, Arain ZM, Rajput N and Abro SH (2009b). A study  
on bacterial contamination of poultry vaccines. Journal of  
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DECLARATIONS  
Competing interests  
The authors declare that they have no competing  
interests.  
Matthijs MGR, Bouma A, Velkers FC, van Eck JHH and Stegeman JA  
(2008). Transmissibility of infectious bronchitis virus H120 vaccine  
strain among broilers under experimental conditions. Avian  
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Authors' contributions  
Moreno A, Franzo G, Massi P, Tosi G, Blanco A, Antilles N, Biarnes M,  
Majó N, Nofrarías M, Dolz R et al. (2017). A novel variant of the  
infectious bronchitis virus resulting from recombination events in  
Italy and Spain. Avian Pathology, 46:28–35. DOI:  
10.1080/03079457.2016.1200011  
Mohamed Morsi Elkamshishi laboratory design and  
publishing, Haitham.H.Ibrahim collected the data and  
wrote the manuscript, Hanan M. Ibrahim performed the  
laboratory work and analysis of the results.  
OIE (2014). Infection disease in OIE quality standard and guidelines for  
veterinary laboratories (world organization for health).  
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