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Salmonella Infection in Calves

Salmonella Infection in Calves

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Published by Mihalca Elena

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Published by: Mihalca Elena on Jan 04, 2013
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SALMONELLA
INFECTION INCALVES: VIRULENCE PROTEINS AND ITSIMMUNOGENIC PROPERTIES
Seleim, R. S.*; Sahar, R. Mohamed; Novert, M. Hafez ; andGobran, R.
 
A.
Bacteriology Department, Animal Heahth Research Institute, Doki, Cairo,Egypt. *Corresponding author
 
SUMMARY
Faecal samples collected from calves suffering from diarrhoea revealedthe isolation of 
Salmonella
in 17.5% of the cases, while in the contactapparently healthy calves the incidence was 3.4%. Four
salmonella
serovares were elucidated namely, S. typhimurium (7.3% and 1.7%),S. dublin (5.1% and 0.8%), S. enteritidis (2.9% and 0.8%) and S.anatum (2.2% and 0%) from diarrhoic and apparently healthy calvesrespectively. SDS-electrophoretic analysis of the outer membraneprotein (OMP) revealed common antigen protein bands especiallybetween
S. dublin
and S. enteritidis, due to the greater similarity intheir antigen structure. All serovars showed intense protein bands inthe range from 20K to 45K. In the Western blot analysis, serumantibodies from calves infected with S. typhimurium (serogroup B)reacted with protein bands at the range of 17K, 24K, and 31K. TheOMP of the two serovars S. dublin and S. enteritidis (both serogroupD1) reacted relatively similar in Western blot with the antiseracollected from calves infected with their corresponding serovars. Twoprotein bands were characteristic for S.dublin and S. enteritidis, 14.4Kand 24K. Only one protein band, 24K from the blotted OMP of S.anatum (serogroup E1) reacted with serum from infected
calvesinfected with that serovar. Using the heterologous serum in the Western blotanalysis gave weaker results than the homologous serum.
ELISA
results detected the presence of serovar specific antibodies,
 
S.typhimurium ELISA detected 10.9% and 4.3%, S. dublin ELISAdetected 7.3% and 2.6%, S. enteritidis ELISA detected 5.1% and1.7%, while S. anatum ELISA detected 2.9% and 0.9% of the serumsamples collected from diarrhoic and apparently healthy calvesrespectively. It could be concluded that
Salmonella
OMP were major
 
immunogenic antigens that could be used in ELSA or Western blot todetect and monitor
Salmonella
infection in calves.
INTRODUCTION
Salmonella
infections in calves continue to be a major problemworldwide. Substantial economic losses were manifested throughmortality and poor growth of infected animals as well as the hazard of transmitting food poisoning to humans. Many outbreaks of 
salmonella
infections has been reported world wide which encountered the mostfrequently isolated serovars as S. typhimurium, S. enteritidis, S.anatum S. newport, S. cerro, S. montevideo, S. agona and S. dublinwhich was considered the major host-adapted
salmonella
for cattle(Mitz et al. 1981, Konrad et al. 1994; Ritchie et al. 2001 and Veling etal. 2002).The infection was always aggravated by poor hygienic conditions andinadequate nutrition and young calves were most susceptible toinfection due to their immature immune responses, undevelopedmicroflora in their gastrointestinal echo-system and the permanentexposure to the source of infection from the environment and theirdams. Calves suffering from acute clinical salmonellosis may shed 10
8
-10
10
 
Salmonella
 /gram faeces and animals recovering from the infectionmay continue to be shedders of the organism for 2-12 weeks postinfection. Shedding
Salmonella
by animals without clinical signs maybe associated with chronic
Salmonella
infection (carrier),convalescence after acute infection or a recent starting colonization(Smith et al. 1989, Galland et al. 2000, Roy et al. 2001 and Radke etal. 2002).Unlike the intermittent shedding of 
Salmonella
in the faeces, antibodylevels do not fluctuate greatly on daily basis. Sero-conversion wasused in many countries as a tool for
Salmonella
surveillance andcontrol in cattle, pig and poultry industry at slaughter to identifyinfected flocks as a regulatory procedures for food safety and securityprogram. Application of ELISA can provide information about theinfection status of the animal and the flock, moreover the repeatedtesting of animals differentiates those recently infected calves(increasing antibody titre) and those convalescent ones (decreasingtitres) from
salmonella
carriers, which would have relatively constanttitres (Smith et al. 1989; Nielsen et al. 1995 and Veling et al. 2002).
 
In order to control and stop the losses incurred by
Salmonella
in calvesformalin-killed
Salmonella
bacterins has been used commercially in thefield in many areas of the world (Wray and Sojka 1984; Roden et al.1992), Yet, further investigations has to be conducted on the outermembrane protein (OMP) and their immunogenic potentials as a firststep for potent vaccine production.The objective of this study was to investigate the infection of calveswith
salmonella
in certain localities, as well as to evaluate theantigenicity of the outer membrane proteins (OMP) by Western blotand ELISA as a preliminary phase for vaccine production.
MATERIALS AND METHODS
Specimens
A total of 253 faecal samples were collected from 137 diarrheic calvesand 116 contact apparently healthy calves aged 1month-1year. At thesame time 253 blood samples were collected from the previouslymentioned animals in order to correlate the incidence of 
salmonella
tothe sero-conversion of each individual animal.Samples were collected during the period of one year from June 2001to June 2002 from calves aged (1month - 1year) at governmental andprivate farms in Giza, Kafr El-Sheikh and Dakahlea governorates.(buffaloe calves were not included in this study).Specimens were transferred to the laboratory in ice box with minimumdelay, where faecal samples were examined bacteriologically for
Salmonella
isolation and identification. Blood samples were allowed toclott, then centrifuged at 700xg for 15min. Sera were collected andstored in frozen aliquot’s until used in the serological tests.
Salmonella
isolation and identification
Faecal samples were cultured into tetrathionate broth and RappaportVassiliadis broth, incubated at 37°C and 42°C respectively for 24hr.Loopful from these broth cultures were then streaked onto Mac Conkeyand S.S agar plates, incubated at 37°C for 24-48hrs and
Salmonella
suspected colonies were identified morphologically, then biochemicallyby the API 20E Kit system (BioMereaux, France) and serologicallyaccording to the Kauffman-White Scheem by slide agglutination test

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