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Microsoft word - antimicrobial susceptibility reaction of pasteurella multocida and.doc

The Sudan J. Vet. Res. (2011), 26: 83-86.
With 2 tables in the text.
Antibiotics Susceptibility Reaction of Pasteurella multocida and
Mannheimia haemolytica
to Selected Eight Antibiotics
Sabiel, Y. A1; Musa, M. T1. and Hadya E Ahmed2 (1)Veterinary Research Institute, P.O. Box 8067 (Al-Amarat), Khartoum, Sudan. E. mail: sabiel- (2) Animal Resources Research corporation , Khartoum, 8 (Mannhaemia haemolytica) مدل ةلاحلا ةيمھنامل ا نم ةلوزعم 1 نم اھلزع مت تلاوزعملا هذھ ر . شتنملا صرقلا رابتخ إةطساوب ةيبوركيملا تاداضملا نم عاونا هينامثل (Pasteurella multocida) ةكتافلا , نيساسكولفوربيس : ةيبوركيملا تاداضملا هذھ ل ش . نادوسلا – قرزلاا لينل ا ةيلاوب نيزامدلا ةناخلس نم تعمج ةبھتلم ناض تائر هساسح تدجو تلاوزعملا لك ن . يلكيسارتتو لوزازكوميارتوك ن . زازكوميارتوك و نيسيماتنجل مواقم اھمظعمو م The antimicrobial susceptibility reactions of 12 Mannheimia haemolytica and 8
Pasteurella multocida isolates from pneumonic sheep ,collected from Al-Damazin
slaughterhouse, the Blue Nile State, Sudan, were evaluated by disk diffusion method. Each
isolate was tested against ciprofloxacin, ceftriaxone, cefuroxime, chloramphenicol, ampicillin,
gentamicin, co-trimoxazole and tetracycline. All isolates of the two species were sensitive to
ciprofloxacin, ceftriaxone, cefuroxime and 90% of them were sensitive to chloramphenicol and
ampicillin. Seventy - five per cent (9/12) of M. haemolytica
isolates were resistant to gentamicin
and co-trimoxazole. Fifty per cent (4/8) of P. multocida
isolates were sensitive to tetracycline
whereas 87.5% (7/8) of them were resistant to gentamicin and 62.5% (5/8) to co-trimoxazole.

Respiratory diseases of sheep are spreading worldwide and in Sudan, incur significant economic losses. Bacterial and viral interactions with other predisposing factors play an important role in the occurrence of the diseases (Adlam, 1989). Members of the family Pasteurellaceae that are present in the mucous membrane of many animal species are opportunistic pathogens and some of them, including P. multocida and M. haemolytica cause diseases in animals and man (Zurlo, 2000). Antimicrobial agents are widely used for the treatment of ovine respiratory infections. During acute infections and outbreaks of infectious diseases in groups or flocks, it is important to use effective antimicrobial agents as early as possible. In acute cases aspirin, fluixin, or ketoprofen may be used for short duration in conjunction with antibiotics therapy to avoid gastric ulceration or renal complications (Kahn and Lane, 2005). This empirical treatment is generally based on knowledge of the resistant pattern of the different bacterial pathogens to antimicrobial agents used in the particular animal species (Hendricksen et al, 2008). Resistance to some antibiotics is frequent in P. multocida and M. Haemolytica, although it is less common in other pasteurellae (Carter and Wise, 2004). Antimicrobial sensitivity tests are important; they are usually carried out to determine antibiotic sensitivity for effective treatment of bacterial infection in vivo. The effectiveness of antibiotic therapy is based on knowledge of the aetiological agent and its relevant antibiotic sensitivity (Wikipedia, 2010). The objectives of this study were to determine the antibiotic sensitivity of P. Multocida and M. haemolytica isolated from pneumonic lungs of sheep and to evaluate the efficacy of the present antibiotics used for the treatment of pneumonia especially pneumonic pasteurellosis in sheep. Materials and Methods
The Kirby-Bauer agar diffusion method (Murray et al, 2003) was used to study antibiotic sensitivity reactions of twelve M. haemolytica and eight P. multocida isolates. Four morphologically similar discrete colonies of each isolate that were cultured onto Blood Agar (Oxoid, CM 271) were selected, picked up with a wire loop, transferred to a test tube containing 4 ml normal saline (0.85%) and incubated for two hours at 37˚C. Each suspension
was adjusted with normal saline to 0.5 McFarland standards (Wikipedia, 2009). They were
swabbed and streaked at three directions over the entire surface of the agar plate. Thereafter a
disc (Multidisc G. X1-minus, Himedia laboratories, India) impregnated with the antibiotics
was picked and placed onto the dried Muller-Hinton Agar plate with a sterile forceps and
pressed down gently to ensure even contact with the medium. The plates were incubated in an
inverted position for 18 h at 37˚C. The sensitivity zones were measured; faint growth and tiny
colonies were ignored. The results were recorded according to Patrick standards (Murray et
al, 2003) as sensitive (S), intermediate (I) or resistance (R) (Table 1).
Table 1: Interpretive standards for disk diffusion method.
Antibiotic Sensitive Intermediate Resistant Unit in mm Chloramphenicol >18 13-17 <12 mm. All isolates of both bacterial species were sensitive to ciprofloxacin, ceftriaxone, cefuroxime and 90% of them were resistance to chloramphenicol and ampicillin. Seventy-five (9/12) of M. haemolytica isolates were resistant to gentamicin and co-trimoxazole. Fifty percent (4/8) of P. multocida isolates were sensitive to tetracycline and 87.5% (7/8) and 62.5% (5/8) were resistant to gentamicin and co-trimoxazole, respectively (Table 2). Table 2: Antimicrobial drugs susceptibility of 12 M. haemolytica and 8 P. multocida
isolates from pneumonic lungs of sheep.

Antimicrobial Susceptibility of Pasteurella multocida and Mannheimia haemolytica Key: Isolates from No. 1 to12=M. haemolytica; from 13 to 20=P. multocida;
C=chloramphenicol; A= ampicillin; TE=etracycline; G= gentamycine; Co=co-trimoxazole
Ci=ceftrixone; Cu= cefuroxime; Cf= ciprofloxacin; S= sensitive; R=resistant;

Antimicrobial sensitivity tests to M. haemolytica and P. multocida have been practised in
many countries. Monitoring drug susceptibility of these bacterial species is essential to
determine resistance development (Post et al, 1991; Watts et al, 1994). The antibiotic
sensitivity profiles obtained are regionally different; therefore their continuous monitoring in
each region is important to select an effective antibiotic drug for treatment of sheep
pneumonia. An increase in the resistance against antibiotics in both organisms was reported in
recent years (Welsh et al, 2004; Hendricksen et al, 2008). In the present study, ciprofloxacin,
ceftriaxone, cefuroxime, chloramphenicol and ampicillin were found most effective
antibiotics against both P.multocida and M. haemolytica. However, P. multocida and M.
showed minimal susceptibility to gentamycin and co-trimoxazole. These results
are similar to those of Verna, or al (2006), Berge et al (2006) and Katsuda et al.
(2009) who reported that all M. haemolytica and P. multocida strains were sensitive to
ceftriaxone, cefuroxime and chloramphenicol.
Our findings are in agreement with Hendricksen et al (2008) who reported an increased resistance of M. haemolytica to ampicillin and tetracycline. Post et al (1991) reported that 90% of P. multocida isolates were moderately susceptible to ampicillin where M. haemolytica isolates were resistant to ampicillin. In a recent study, Milan et al (2009) have found that multi - resistance of P. multocida to multiple clinically used antibiotics is mediated by co-existence of small plasmids. In conclusion, the selection of antibiotic for effective therapy and prevention of increasing multi - drug resistance should be prudently performed on the basis of antimicrobial susceptibility tests, to treat cases of pneumonia in sheep with the suitable antibiotic effectively. Acknowledgements
We wish to thank Ministry of Animal Resources Khartoum State, Animal Resources Bank, Director of Port Sudan Veterinary Research Laboratory and Director of Animal
Resources, Blue Nile State. Thanks are extended to the Director of VRI and Director General
of ARRC for permission to public the article.
Adlam, C. (1989). The structure, function and properties of cellular and extracellular
components of Pasteurella haemolytica, In: C. Adlam and J. M. Rutter, (eds) Pasteurella and Pasteurellosis. Edn edition Edn. Academic Press London , UK. Pp : 75-92. Berge, A. B.; Sischo, W. M. and Craigmill, A. L. (2006). J.A.V.M.A, 229
(8): 1279 – 1281
Carter, G. R. and Wise, D. J. (2004).
Essentials of Veterinary Bacteriology and
. 6th edn., Iowa State Press, USA. Pp. 149–154.
Hendricksen, R. S.; Mevius, D. J.; Schroater, A.; Teale, C.; Meuier, D.; Butaye, P.;
Franco, A.; Utinane, A.; Armando, A.; Moreno, M.; Greko, C.; Stark, K.;
Berghold, C.; Myuyniemi, A.; Wasyl, D.; Sundle, M. and
Aarestrup, F. M.
Acta Vet. Scand. 50(22) .10 1186/1751.
Kahn, C. M. and Lane, S. (2005): The Merck Veterinary Manual, 9th edn. Publisher: Merck, (2005)
Katsuda, K.; Kohmoto, M.; Mikani, O. and Uchida, I. (2009). Vet. Microbiol., 139: 74-79.
Milan, A. S.; Escudero, J. A.; Belen, G.; Laura, H.; Nerea, M.; Montserrat, L.; Lucas,
D.; Bruno G. and Zorn, B. G. ( 2009).
Antimicrobial Agents Chemotherap. 53 (8):
Murray, P. R.; Baron, E. J.; Jorgensen, J. H.; Pfaller, M. A. and Yolken, R. H. (2003) .
Manual of Clinical Microbiology. 8thedn. American Society for Microbiology (ASM) ,Washington, USA, Pp. 1108-1127. Post, K. W.; Cole, N. A. and Raleigh, R. H . (1991). J. Vet. Diagn. Invest., 3: 124-126.
Verna or Vera, Y. A.; Rodriguez-Ferri, E. F; de la Fuente, A. J. and Gutierrez–
Martin, C. B. (2006). Am. J. Vet. Res. 67(4): 663-668.
Watts, J .L.; Yancey, R. J.; Salmon, S. A. and Case, C.A,(1994): J. Clin. Microbiol.,
Welsh, R. D.; Dye, L.B.; Payton, M. E. and Confer, A.W. (2004). J. Vet. Diagn.
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Wikipedia. (2009). http:// en.Wikipedia. org/wiki/McFarland Standards. Wikimedia
Wikipedia. (2010). Antibiotic sensitivity.
Zurlo, J. J. (2000). Pasteurella species. In : G. L. Mandel, R. F. Douglas and J. E. Bennet.
(eds.). Principles and practice of infectious diseases, 5th edn. Churchill Livingstone,
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