ISOLATION AND IDENTIFICATION OF BACTERIA FROM KIDNEY LESIONS IN BROILERS IN SULAIMANIA PROVINCE

ISOLATION AND IDENTIFICATION OF BACTERIA FROM KIDNEY LESIONS IN BROILERS IN SULAIMANIA PROVINCE

AMEEN N.A.^*; AREF E.D.^**; AL-OBAIDI R.M.^*** and RAOOF H.H.^****

^*Dept of Histopathology, College of Veterinary Medicine, Sulaimania University, Kurdistan Region, Iraq

^**Dept. of Microbiology, College of Veterinary Medicine, Sulaimania University, Kurdistan Region, Iraq

^***Dept. of Basic Sciences, College of Veterinary Medicine, Sulaimania University, Kurdistan Region, Iraq

^****Department of Animal Health, Sulaimania Veterinary Directorate, Ministry of Agriculture and Water Recourses, Kurdistan Region, Iraq

Email: nzarameen@yahoo.com                                                                               Assiut University Email: www.aun.edu.eg

INTRODUCTION

Bacteria are pathogenic living organisms but, not all bacteria are detrimental to poultry health. In fact, many bacteria are beneficial and necessary for such processes as food digestion and manufacturing of some dairy products. Classification of bacteria into species is done so disease producing organisms may be separated from those that are harmless or beneficial, (Berry, and Whitenack, 2011).

Bacterial nephritis usually occurs when bacteria enter the kidney secondary to systemic disease through the renal arteries or the renal portal system (Speer, 1997 and Phalen et al., 1990). A wide range of bacteria has been reported to cause bacterial nephritis including Enterobacteriaceae, Pasteurella spp, Pseudomonas spp, Streptococcus spp, and Staphylococcus spp (Speer, 1997, Schmidt et al., 2003, Phalen et al., 1990 and Lierz, 2003). Listeria monocytogenes has been reported (Schmidt et al., 2003), whereas Erysipelothrix rhusiopathiae has been reported in quail and chicken (Schmidt et al., 2003 and Mutalib et al., 1995). Mycobacterium avium can, rarely, cause renal lesions (Schmidt et al., 2003 and Sato et al., 1996).

Among bacterial infections, colibacillosis is a worldwide major cause of morbidity and mortality in poultry and responsible for significant economic losses to the poultry industry, (Stordeur, and Mainil, 2002) they also reported that the causative bacteria, avian pathogenic Escherichia coli (APEC), induce various syndromes including respiratory tract infection (airsacculitis), acute colisepticemia, salpingitis, and cellulitis. The most common form of colibacillosis occurs in 3- to 10-week old chickens. It is characterized by an initial respiratory infection usually induced by mycoplasmal and/or viral infections and enhanced by a high level of ammonia in poultry houses. The disease evolves as a systemic infection (perihepatitis, pericarditis, and septicaemia) due to the invasive abilities of the Escherichia coli strains (Dho-Moulin, and Fairbrother,1999), hence, can infect both kidneys and cause damage to the kidneys and impair its function (Al-Hiyali et al., 2005).

Successful control of bacterial diseases entails isolating and identifying disease-producing species, if present, and preventing multiplication and spread of the organism within the chick's body or to other chicks, also antibacterial resistance has profound clinical implications. The objectives of the present study are to isolate and identify bacterial species infect broiler chicks kidney, furthermore to investigate antibacterial susceptibility and describe the changes in the kidneys by means of histopathology.

MATERIALS and METHODS

A total of ninety seven broiler chicks suffering from nephritis were collected from different flocks in Sulaimania region at different dates. Samples of Affected kidneys were cultured on 5% blood agar base, Mannitol salt agar, MacConkey agar plates (Quiun et al., 2002) the inoculated plates were incubated at 37c°for 24-48 hours. Primary cultures were evaluated by visual examination of the morphology of the bacterial colonies and were sub cultured for further examination. The identification of the isolated colonies was performed using standard bacteriological and biochemical procedures as described by carter and Cole (1990) and barrow and felthem (1993) and conformed by API 20E system  (biomerieux, Marcy-1 Etoile, france) as per manufactures directions.

Antibacterial susceptibility tests performed to detect the susceptibility of the  isolated bacteria to antibiotic which included, Doxycycline 30mg, Ciprofloxacin 5mg, Neomycin 30mg, Ampicillin 10mg, Chloramphenicol 30mg, Co-trimoxazole 25mg (England) (Brown, 2005).

To study histopathological changes, 1 cubic centimeter size were obtained from the affected kidneys samples of all chicks and kept in 10% neutral buffered formalin for at least 24 hours. After that, these samples were exposed as described by (james, 1976) for dehydration, clearing, paraffin embedding, blocking, sectioning, mounting and staining with Hematoxylen and Eosin stains  and were examined by light microscope.

RESULTS and DISCUSSION

bacteriological and biochemical tests results showed 89.69 % (87) of isolates were E.coli, 9.27 % (9) isolates were Proteus milabilis and 1.03 % (1) isolates was Klebsiella pneumonia as shown in table (1). The isolates were confirmed by The API 20E test kit (biomerieux, Marcy-1 Etoile, france).

Table 1: Shows bacterial isolates from kidneys samples.

The isolation results of the present study revealed high percentage of E.coli which agree with results mentioned by (Al-Hiyali et al., 2005) and (Sokker    et al., 1998), the high percentage of the E.coli isolates (89.69%) (Table 1), confirmed the importance of this bacteria for inducing the lesions in kidneys of broiler chicks which is identical to the results of (Al-Hiyali  et al., 2005) and (Al- Sultan et al., 1998) in other animals (calves) and also resemble the result revealed by (Al Rajab and Al qazzaz, 1984) in urinary tract infection of human; the explanation of this results is thought due to the absence of lymph nodes and the presence of renal and hepatic portal systems which increase the risk of systemic or gastrointestinal microbes affecting the kidney (Lierz, 2003 and Lumeij 2000), also bacterial nephritis usually occurs when bacteria enter the kidney secondary to systemic diseases through the renal arteries or the renal portal system, Rarely, bacteria ascend the ureters secondary to conditions such as chronic cloacitis (Speer, 1997, Lierz 2003 and Phalen et al., 1990).

Other bacteria including Proteus mirabilis and Klebsiella pneumonia were isolated out of several kidneys samples but in a lower proportion, mentioned in Table (1), these results were similar to results reported by (Al-Hiyali et al., 2005) in chicken  and also similar to results showed by (Quinn et al., 1997) in cattle, buffalo and dog.

In vitro Antimicrobial susceptibility test of the most frequent bacterial Isolates (E.coli isolate) were susceptible to a (Ciprofloxacin and Co-trimoxazole) but showed resistance to (Doxycycline, Ampicillin and -Chloramphenicol) as shown in Table (2) and figure (1), The results of susceptibility testing are identical with those mentioned by Carter and Cole (1990), Carter et al. (1995).

Table 2: Shows Antimicrobial susceptibility test of E.coli isolate in the present study.

(S: sensitive, R: resistant, I: intermediate)

Figure 1: Antimicrobial susceptibility test of E.coli isolate shows susceptible to a (Ciprofloxacin and Co-trimoxazole) but resistant to (Doxycycline, Ampicillin and -Chloramphenicol)

The histopathological examination of affected kidney included the enlargement of epithelial cells of renal tubules with pale cytoplasm, blood capillaries were engorged with large numbers of Red blood cells in both renal glomeruli and interstitial tissues, and congestion of blood vessels in addition to reduction in the size (atrophy) of renal corpuscles as shown in figure (2), the results of histopathological examination of the current study are identical with those mentioned by (Sokker et al., 1998).

Figure 2: Congestion of blood vessels and atrophy glomeruli (400X)

In conclusion, this work revealed that the E.coli infections are the most real important causes of kidneys lesions leading to economic losses in the broiler chickens thus prevention and control of E.coli infections in broiler farms are very important.

ACKNOWLEDGMENT

This work was supported by College of Veterinary Medicine, University of Sulaimania.

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