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Antibiotic Susceptibility of Major Bacteria Cause Ovine Mastitis in River Nile State, Sudan

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Antibiotic Susceptibility of Major Bacteria Cause Ovine Mastitis in River Nile State, Sudan Imperial Journal of Interdisciplinary Research (IJIR) Vol-3, Issue-1, 2017 ISSN: 2454-1362, http://www.onlinejournal.in Antibiotic Susceptibility of Major Bacteria Cause Ovine Mastitis in River Nile State, Sudan Fadlalla E.2 ,1٭Abubaker A. El Ayis 1Department of Medicine, College of Veterinary Medicine, Bahri University (Sudan). 2Department of Biochemistry, College of Veterinary Medicine, Bahri University (Sudan). Abstract: In this study a total of 40 milk samples Procaine Penicillin and Cloxacillin. 50.0% of positive for California Mastitis Test (CMT) were Klebsiella pneumoniae isolates were sensitive to collected from ewes in the three localities of River Ciprofloxacin, Gentamycin and Kanamycin. All Nile State (Atbara, Barbar and El Damer). Samples strains were resistant to Ampicillin, Cephalothin, were submitted for bacteriological examination. Chloramphenicol, Erythromycin Procaine The aerobic bacteria isolated and identified from Penicillin, Cloxacillin and Tetracycline. All ewes’ milk samples were 19 Staphylococci Enterobacter spp. Isolates were sensetive to (21.0%), 9 Streptococci (9.9%), 5 Echerichia coli Cephalothin and Chloramphenicol, 66.7% to (5.6%), 4 Corynebacterium bovis (3.3%), 3 Ciprofloxacin, Tetracycline and Gentamycin and Enterobacter spp. (3.3%), 2 Klebsiella pneumoniae all isolates were resstant to Ampicillin, spp, (2.2%) and 2 Enterococcus faecalis (2.2%). Erythromycin, Kanamycin, Procaine Penicillin and All isolates were subjected to sensitivity tests using Cloxacillin. 10 antibiotics in use for treatment of mastitis in Sudan. 89.5% Staphylococcal isolates were I. INTRODUCTION sensitive to Ampicillin, 73.3% to Cephalothin, 68.4% to Ciprofloxacin and Kanamycin, 63.2% to Mastitis is one of the more common health Gentamycin, 57.9% to Chloramphenicol, 26.3% to problems affecting sheep and goats. The bacteria which are known to cause mastitis in cows, sheep Tetracycline, 15.8% to Procaine Penicillin and and goats are Streptococcus sp., Staphylococcus Cloxacillin and 5.3 to Erythromycin. 88.9% of sp., Pasteurella sp., and coliforms, such as E. coli. sterptococcal isolates were sensetive to Ampicillin, The most commonly isolated CNS species in 66.7% to Gentamycin, 55.6% to Ciprofloxacin and persistent subclinical in goats and sheep are Kanamycin, 44.5% to Cephalothin, 33.4% to Staphylococcus epidermidis, S. caprae, S. Chloramphenicol, 22.2% to Procaine Penicillin simulans, S. chromogenes and S. xylosus [1, 2, 3]. Other pathogens such as Streptococcus spp., and Cloxacillin and 11.1% to Erythromycin and all Enterobacteriaceae, Pseudomonas aeruginosa, strains were resistant to Tetracycline. 80% of Mannheimia haemolytica, Corynebacteria and Echerichia coli isolates were sensitive to fungi can produce mastitis in small ruminants, but Ciprofloxacin, 60.0% to Ampicillin, Cephalothin, occurrence rates are lower. In addition, severe Chloramphenicol and Tetracycline, 40.0% to cases of mastitis related to incorrect preventative Gentamycin and Kanamycin, 20.0% to strategies have been attributed to the pathogens Erythromycin and Cloxacillin and all strains were Aspergillus fumigatus, Serratia marcescens, P. aeruginosa or Burkholdelia cepacia [4, 5, 6, 7]. [8] resistant to Procaine Penicillin. All isolated many bacteria from cases of subclinical Corynebacterium bovis isolates were sensitive to and clinical mastitis. These include: S. aureus, S. Gentamycin, 66.6% to Cephalothin, 33.3% to epidermidis, Str. agalactiae, Str. ubris, E. coli, Ps. Ampicillin, Kanamycin, Tetracycline and aeruginosa and K. pneumoniae. Enterobacter spp Cloxacillin and all strains were resistant to were found to cause bovine mastitis [2]. Chloramphenicol, Ciprofloxacin, Erythromycin, Corynebacterium spp alone were found to be and Procaine Penicillin. All Enterococcus faecalis associated with clinical mastitis [3]. The choice of antibiotic to be used can be decided isolates were sensitive to Ciprofloxacin and by antibiotic sensitivity test [9]. [10] studied the Gentamycin. 50.0% to Cephalothin, antibiotic sensitivity of 281 S. aureus strains Chloramphenicol, Kanamycin and Tetracycline. All isolated from bovine milk in U.S.A, using 21 strains were resistant to Ampicillin, Erythromycin, antimicrobial agents. They reported that more than Imperial Journal of Interdisciplinary Research (IJIR) Page 908 Imperial Journal of Interdisciplinary Research (IJIR) Vol-3, Issue-1, 2017 ISSN: 2454-1362, http://www.onlinejournal.in 90% of S. aureus isolates were sensitive to II. MATERIALS AND METHODS Bacitracin, Cephalothin, Chloramphenicol, Area of the study Cloxacillin, Erythromycin, Gentamycin, A total of 40 milk samples positive for CMT were Kanamycin, Lincomycin, Neomycin, collected from goats in River Nile State. Milk Nitrofurazone, Oleandomycin, Tetracycline and samples were collected from Barbar, Omer Amir Vancomycin. Ampicillin, Penicillin and Polmyxin farm, El Damer Vet Hospital, Abdelghafarm B were ineffective against the majority of S. aureus farm,ood safety Center, Atbara vet hospital and isolates. [11] studied the antimicrobial Akram farm (table 1). susceptibility of coagulase–negative staphylococci Sampling procedure isolated from bovine mastitis in Argentina and no Before collection of milk samples from the tested resistance was detected for Gentamycin, cows, the udder was thoroughly cleaned with soap Cephalothin, Ampicillin, Cloxacillin, Erythromycin and water, rubbed dry, and the teat area was rubbed and Clindamycin. The results also indicated that thereafter with a piece of cotton soaked in 70% coagulase–negative staphylococci isolates in alcohol . The first stream of milk was discarded. Argentina exhibited the highest degree of The California Mastitis Test was directly applied resistance to Penicillin of all antimicrobial agents for quarter's milk and samples were collected from tested. [12] found that Streptococcus agalactiae positively reacted milk into sterile bottles.The and S. intermedius isolates were resistant to collection of samples was at (2-5) pm. The tetracycline (61.2%), followed by lincomycin collected samples were put in ice box containing (43.2%), gentamycin (35.3%), oxacillin (34.3%), ice and transported to the laboratory. In most cases and erythromycin (28.6%). Cephalothin and the time between collection and arrival to the penicillin were the only antimicrobial agents to laboratory was 1-2 hrs. In the laboratory mastitic which most of the streptococci (92%) were milk samples were kept in a deep-freeze. All susceptible. E. coli showed variation in their samples were examined on the next day. On the susceptibility to various chemotherapeutic agents in next day mastitic milk samples were removed from use for treatment of calf diarrhoea such as the deep–freezer and left on the bench to thaw. Amoxicillin, Ampicillin, Chloramphenicol, Samples were then cultured. Ciprofloxacin, Gentamycin, Kanamycin, Nalidixic Isolation, identification and characterization of acid, Streptomycin, Sulphamethoxazole, and bacterial isolates Erythromycin [13]. [14] reported that E. coli All media (Oxoid media) were prepared and isolates were highly sensitive to Nitrofuran(98%), sterilized according to the manufacturer Colistin sulphate (98%), Gentamycin (97%) and instructions. For the primary isolation of bacteria, a Cotrimoxazole (82%) and resistant to the rest of loop full milk sample was streaked onto blood agar, antibiotics (Streptomycin, Tetracycline, Nalidixic McConkey's agar, and nutrient agar using sterile acid and Ampicillin). Enterococcus faecalis usually wire loop. The cultures were incubated aerobically susceptible to Ampicillin and Penicillin. It can at 37oC for 18-24 hours. Cultures on semi-solid acquire resistance to Vancomycin, usually due to media were examined grossly for colonial van A or van B and canoccasionally produce beta- morphology and haemolysis on blood agar. lactamase [15]. [16] found that coryneform bacteria Whereas, broth media were checked for turbidity, were 100% resistant to penicillin G except change in colour, accumulation of gases in CHO Corynebacterium ulcers (60%). In contrast media and for sediment formation. One half colony Gentamycin was the drug of choice for from each plate was used for performing gram Coryneform. staining. Colonies which showed Gram positive The most sensitive antimicrobial was amikacin and cocci were sub cultured on nutrient agar. imipenem with 62 (86.1%) Klebsiella pneumoniae Purification was based on the characteristics of isolates susceptible to it, followed by gentamicin colonial morphology and smear. This was obtained and tobramycin with 58 (80.6%), ofloxacin with 57 by sub culturing of a typical discrete colony on (79.2%), ciprofloxacin with 55 (76.4%). 30 blood agar plate. Pure cultures were preserved on (41.7%) Klebsiella pneumoniae isolates were slants of blood agar and egg media at 4Co. multiresistant to all the antimicrobial agents used Biological and biochemical identification [17]. Resistance to ampicillin and penicillin in The purified isolates were identified as previously Enterobacter spp. is primarily due to changes in the described [18] and [19]. The identification include: penicillin-binding proteins (PBPs) which decrease Gram’s reaction, presence or absence of spores, the affinity of the PBP target proteins for beta- shape of organism, motility, colonial characteristics lactam drugs. Some enterococci produce on different media, aerobic and anaerobic growth, aminoglycoside-modifying enzymes that confer sugars fermentation ability and biochemical tests highlevel aminoglycoside
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