Veterinary Science and research Volume 1: 1 Vetry Sci Rech 2019

Advanced Bacteriological Studies on Bumblefoot in Broiler with Some Clinicopathological Alteration

Fatma M Youssef1* 1Department of Clinical Pathology, Animal Health Research Abdelmohsen A Soliman1 Institute, Ismailia, Egypt Ghada A Ibrahim2 2Department of bacteriology, Animal Health Research Institute, Hend A Saleh2 Ismailia, Egypt

Abstract Article Information S. aureus is responsible for Bumblefoot and septic arthritis in broilers Article Type: Research and layers. The present work is designed to investigate and evaluate Article Number: VSR101 the most common bacterial causes of bumblefoot disease and their Received Date: 08 February, 2019 hematological, biochemical and immunological effects in broiler chicks. Accepted Date: 13 February, 2019 For bacteriological examination, one hundred and twenty foot swabs were Published Date: 15 February, 2019 collected from diseased chicks and investigated for the bacterial causes. Also, blood samples were collected from the same cases for hematological, *Corresponding author: Fatma Mohamed Yousseff, biochemical and immunological analysis. In this study, S. aureus was Department of Clinical Pathology, Animal Health Research isolated in 45.8% of diseased broilers. It was isolated either in pure form Institute, Ismailia, Egypt. Tel: +01025250063; Email: fatmayousseff(at)yahoo.com (18.18%); or in a mixed form with other species like: E.coli (58.18%), Proteus mirabilis (14.67%) and Pseudomonas aeruginosa (9.1%). Molecular characterization of Coa and Spagenes of S. aureus isolates Citation: Youssef FM, Soliman AA, Ibrahim GA, Saleh HA (2019) Advanced Bacteriological Studies on the highest sensitive antibiotic drug against the isolated species followed Bumblefoot Infections in Broiler Chicken with Some Clinicopathological Alteration. Vetry Sci Rech Vol: 1, Issu: were detected with PCR in 70% and 80%, respectively. Levofloxacin was 1 (01-09). most recovered isolates exhibited moderate to high resistance against by gentamycin, ciprofloxacin and amoxicillin antibiotics. However, Copyright: © 2019 Youssef FM. This is an open-access trimethoprim, oxaciliin, enerofloxacin, tetracycline and erythromycin. article distributed under the terms of the Creative Commons concentration and PCV in the diseased broiler chicks indicated anemia. Attribution License, which permits unrestricted use, InFor addition, hematological leucocytosis, parameters, lymphocytosis a significant and decrease monocytes in RBCs is in count,naturally Hb distribution, and reproduction in any medium, provided the original author and source are credited. uric acid, creatinine, gamma, alpha globulin, IL-6 and TNF-α levels were infectedrecorded groups in naturally were recorded.infected chicks. Also, a Total significant protein increase and albumin in AST, levels ALT, ofwere total significantly leukocytic decreased.count, lymphocyte The levofloxacine and monocyte treated as groups compared revealed with significant increase in RBCs, Hb, and PCV as well as decreased number representsinfected groups. a promising Levofloxacine candidate also improved to treat the levelbacterial of liver infections. function, Staphylococcalkidney function, infections IL-6 and TNF.are higher It could in bebroilers concluded than that,other levofloxacin organisms. Therefore, the epidemiological importance and good hyagenic measures to humans. should be taken in this field to minimize the transmissible diseases of Keywords: S. aureus Haematological, AST, Antioxidant, IL6, TNF. , Isolation, PCR, Spa gene, Coagulase, Levofloxacin, Introduction degenerative condition of the avian foot. It usually occurs with age (14 -70Bumblefoot days), but most is a cases broad occurred term thataround includes 35 days any old inflammatory[1]. It may range or from a very mild redness or swelling to chronic, deep seated abscesses and bony changes [2]. www.innovationinfo.org S. aureus is one of the important diseases of Sampling poultry that are common in commercial broilers and layers. Bacteriological swabs from (pus or infected lesions) It causes mortality rate (0-15%) and reduces production diseased cases showing signs of arthritis or Bumblefoot or performance of . S. aureus is a normal inhabitant of the leg affections also, lung and liver organs (from those suffering skin and upper respiratory tract in [3]. In poultry, from respiratory signs) were collected for bacteriological it has been implicated in arthritis, osteomyelitis, synovitis, examination. In addition, blood samples (anticoagulant cellulites, dermatitis, endocarditis, septicemia, wound blood and serum) were collected from broiler chicks (20 infection, ophthalmitis and omphalitis [4]. for each)for hematological, biochemical and immunological This disease is most commonly caused by S. aureus [5] studies. Also, after 5 days of treatment, another twenty blood and sometimes involved with E.coli [6] which is of veterinary samples from the treated broiler chicks were collected. importance in broilers. In such birds, the most common Bacteriological examination Isolation and identification of bacterial microorganisms: Mostform bacterial of infection pathogens involves could tenosynovitis play a role ( in the incidence of All sample swabs and organ samples were cultured onto oftendon respiratory sheaths) disease and arthritis in domestic of the hockpoultry and species stifle joints [8]. The[7]. nutrient broth, incubated aerobically at 37°C for 24 hours, respiratory tract infections are of eminent importance in the then streaked on blood and macConkey agar plates and manufacture of poultry because of high mortality in poorly re-incubated for 24 h at 37°C. Following incubation, the managed cases [9] which lead to large economic losses [10]. colonies were picked up and sub-cultured till pure growths Hematological alterations were recorded the severity of isolation of S. aureus with streaking method, then incubated. were obtained. Mannitol salt agar media used for the specific concentration and PCV in the affected birds indicate anemia The hemolysin activity of S. aureus and E.coli isolate was ofbacterial microcytic infection, hypochromic a significant [11]. decrease Moreover, in theRBCs biochemical count, Hb Also, EMB (Eosin Methyline Blue) and Pseudomonas F agar examined on sheep blood agar medium plates at 37˚C [16]. change in protein. Also, Hypoalbuminemia was observed as E.coli and wellanalysis as increase recorded of increaseserum uric in ASTacid andand ALTcreatinine and a significant[11]. Pseudomonas species. The purity of the isolated organisms mediawas determined were used foron species the basis specific of detectiontheir morphological, of cultural characteristics and Gram’s staining. Biochemical which has been extensively used for the treatment of bacterialLevofloxacin infections. is a Itmember is known of withthe fluoroquinolone its wide antimicrobial group according to [17]. activity against both Gram-negative (including Pseudomonas identification of the isolated bacterial species also was done Serological identification of the isolates E.coli : E.coli species) and some Gram-positive organisms (including S. aureus). It also has enhanced activity against Streptococcus by using pneumoniae, S. aureus and Enterococcus species, besides its isolatesof polyvalent were biochemicallysomatic antisera, identified for application then were of subjectedthe slide good activity against Mycoplasma and Chlamydia species toagglutination the serological test. identificationThis work was according performed to in [18], the serology [12]. Its pharmacokinetics studies explained its tremendous unit of Animal Health Research Institute (AHRI). tissue penetration, large volume of distribution and relatively short elimination half-life in animal body. Takizawa et al. Congo red Binding Assay [19]: E.coli positive samples streaked on Congo red agar plates to determine the useful in a variety of animal infections including urinary, pathogenicity of E.coli isolates. Each isolate was streaked on (1999) [13] proved that levofloxacin drug was extremely sterile separate plate and kept at 37°C for 24hrs. After 24hrs incubation, the cultures were kept at room temperature pharmacokinetics studies had been reported in birds and respiratory, soft tissues, bones and joints. Levofloxacin for 48 hours. Pathogenic E.coli red positive isolates produced brick red colonies. The non- were identified by Congo layers [14]. The oral administration of levofloxacin(10mg/ pathogenic isolates appeared as colorless after 48 hours in on hematological or biochemical values were observed [14 room temperature. andkg) in15]. broiler Thus, birds the wasaim foundof this safe study as non-significantwas to investigate change the bacterial causes of Bumblefoot disease in broiler farms Antibiotic sensitivity testing of isolated : with molecular detection of some genes as well as the S. aureus, E.coli, Proteus and hematological, biochemical, and immunological changes Pseudomonas species were tested against a panel of nine The identified isolates of accompanied with the disease. Materials and Methods commonly used antimicrobial agents: Levofloxacin (1 µg), Amoxicillin+clavulanic acid (10 µg), Gentamicin (120 µg), History and clinical examination: A total of one hundred Ciprofloxacin (5 µg), Erythromycin (15 µg), Tetracycline and twenty broiler chicks of age (20-35 days) that were method(10 µg), [20]. Enrofloxacin The results (5 µg),were Trimethoprim interpreted according (30 µg) and to reared in broiler farms at Ismailia and Zagazig Governorates theOxacillin criteria (1µg)with recommended the standard by the Kirby–Bauer Clinical and disc Laboratory diffusion in Egypt were used in the present study. They all showed Standards Institute for antimicrobial susceptibility testing the clinical signs of arthritis, leg affections or Bumblefoot disease and about eighty cases accompanied with some respiratory signs. [21]. The susceptibility of identified to isolates resistant to three or more antibiotics were classified as Multidrug Drug Resistance (MDR) strains. Vetry Sci Rech 2019 02 www.innovationinfo.org Table 1: Oligonucleotide primer sequences of coa and spa genes of S.aureus. Immunological studies Target gene Primer sequence (5’-3’) References Coa F ATA GAG ATG CTG GTA CAG G Iyer and Kumosani • (2011) Polyacrylamide gel electrophoresis according to [29] in F GCT TCC GAT TGT TCG ATG C theProtein animal electrophoresis health research institute was done in the biochemistry using SDS- TCA ACA AAG AAC AAC AAA Spa F Wada et al. (2010) department. ATG C The collected serum F GCT TTC GGT GCT TGA GAT TC • Determination of interleukin-6: showed 100% cross-reactivity with the ELISA kit, and

Molecular detection of Coa and Spa genes of S.aureus assays were performed according to the manufacturer’s DNA extraction: it was done for ten selected S.aureus protocol.the limit of serum IL-6 detection was 31.2 pg/ml. ELISA The test was GmbH) were performed at RLQP (Reference Laboratory • Determination of Tumor Necrosis Factor: drawn according to [30]. forisolates Veterinary using theQuality QIAamp Control DNA on Mini Poultry kit (Qiagen, Production) Germany, with Coa, Field treatment: Naturally infected chicks received and Spa genes of S.aureus some manufacturer’s modifications for screening for isolates. Briefly, 200µlofthe sample treatment to farms as the following: Levofloxacin (10 mg/ ofsuspension 100% ethanol was incubated was added with to 10µl the lysate.of proteinase The sample K and was200 askg b.w)the following: [31], was addedGrinding to theof tablets drinking (500 water. mg) Levofloxacin with 10 ml thenµl of lysiswashed buffer and at centrifuged 56 °C for 10 following min. After the incubation, manufacturer’s 200µl distilledwas purchased water fromthen Sanofidrinking Aventis to chicks company, according and prepared to dose of elution buffer provided in the kit. The Oligonucleotide recommendations. Nucleic acid was eluted with 100 µl Primers used were supplied from Metabion (Germany) were under(levofloxacin observation 10 mg/kg after theb.w) end as ofconcentrate treatment. 0.2 ml per os/ chick/once daily) for four successive days. Birds were kept Statistical analysis: The obtained data were statistically listed in (Table 1 and 2). For PCR amplification, the primers analyzed by an ANOVA (one way) variance method were utilized in a 25µlreaction containing 12.5µl of Emerald considering P < 0.05 using MiniTab17© software. The Amp Max PCR Master Mix (Takara, Japan),1µl of each biosystemprimer of 202720 pmol thermal concentrations, cycler. 4.5µl of water, and 6µl tests to compare the means [32]. of DNA template. The reaction was performed in an applied significant differences were taken to Fisher multiple range Analysis of the PCR Products: The products of PCR Results were separated by electrophoresis on 1.5% agarose gel (Applichem, Germany, GmbH) in 1x TBE buffer at room In this study, the observed clinical signs in diseased broiler chicks were: swollen foot pad Figure 1: (1), foot bad of the uniplex PCR products were loaded in each gel slot. A temperature using gradients of 5V/cm. For gel analysis, 20µl (3), inability to stand (4). Post Mortem examination showed dermatitis (2), swelling and inflammation of their hock joint gene ruler 100 bp ladder (Fermentas, Sigma) were used to that 45 out of 120cases showed congestion of liver with clear determinegel pilot 100 the bp fragment DNA ladder sizes. (Qiagen, The gel Germany,was photographed GmbH) and by a gel documentation system (Alpha Inno tech, Biometra) and the data were analyzed through computer software. (Table 1 and 2) Hematologicalstudies: erythrocytic count, hemoglobin concentration, packed cell volume, total leukocytic countDetermination and the differential of theleukocytic total count were determined according to [22]. Serum biochemical analysis: The collected sera were assayed for serum biochemistry. The level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) according to [23] creatinine [24], uric acid [25], total serum proteins [26], albumin [27] and globulin [28]. These parameters were spectrophotometrically assayed by Figure 1: (1), foot bad dermatitis (2), swelling and inflammation of their hock using semi-automated spectrophotometer (Erba-Chem7, joint (3), inability to stand (4). Post Mortem examination showed that 45 out Germany) using commercial kits purchased from (Spectrum, of 120cases showed congestion of liver with clear perihepatitis, congestion of Cairo, Egypt). lung and pericarditis of heart (5 and 6).

Table 2: Cycling conditions and predicted sizes of PCR products for Coa, and spa genes of the isolated strains. Actual cycles (35) °C/sec Final extension °C/ Target gene Initial denaturation Amplified product Size (bp) Denaturation Annealing Extension min Coa 94/10 94/ 60 55/60 72/10 72/10 570 Spa 94/5 94/30 55/30 72/30 72/10 226

Vetry Sci Rech 2019 03 www.innovationinfo.org Table 3: Bacterial causes for affection with arthritis/ Bumblefoot in chicken cases. Total No. of positive isolates bacteria from 120 samples Bacterial species Cases No % S. aureus (pure form) Bumblefoot 10/120 8.33% S. aureus mixed with E. coli 32/120 26. 67% S. aureus mixed with Proteus 8/120 6. 67% species Bumblefoot with respiratory signs S. aureus mixed with P. 5/120 4. 17% aeruginosa Total 55/120 45. 8%

Table 4: The results of Serotyping of isolated E.coli and Congo red binding test. E.coli Serotypes Isolates (32) Congo red binding test were mixed with E.coli (26.67%); eight isolates were mixed liver organs of other cases with respiratory signs: 32/120 O78 10 Positive with Proteus species Pseudomonas

O55 5 Positive aeruginosa (4.17%) as shown in Table 3.

O158 6 Negative (6.67%) and five with O 5 Negative Serotyping of the yielded E.coliisolates revealed that ten 86A isolates were of O , six of O , four O1 4 Positive 78 55 158 86A of O and two ofO as shown in the table (4). With Congo O11 2 Negative 1 11 Total +ve No. of E.coli isolates with Congo red red testing, serotypes, five of Oof O, O and O were, five positive of O while 19 78 55 1 binding test others didn’t give the reaction. The Congo red positive result was indicated by the development of orange or bright red Table 5: Antibiotic susceptibility testing of the isolated species. colonies, while the white color indicated as a negative result. The recovered bacterial isolates (Table 4). Antibiotics S. aureus E. coli Pr. Mirabilis P. aeruginosa (55) (32) (8) (5) The results of antibiotic sensitivity testing of the Levofloxacin (1 µg)) S S S S recovered isolates ofS. aureus, E.coli, P. mirabilis and P. Amoxicillin+clavulanic aeruginosain this study developed highest sensitivity level S I S R acid (10µg) Gentamicin (120 µg) S I S I Ciprofloxacin (5 µg) S I S I erythromycinagainst levofloxacin. exhibited Gentamycin, showed ciprofloxacin, varying degree amoxicillin, from Erythromycin (15 µg) R I S R intermediatetrimethoprim, to oxaciliin,resistant as enerofloxacin, showing in Table tetracycline 5. and Tetracycline (10 µg) R S S I Molecular characterization of coa and spa genes of S. Enrofloxacin (5 µg) R I I R Trimethoprim (30 µg) R R R R aurues isolates showed that 70% and 80% of the tested Oxacillin (1 µg) R R R R

size:isolates 570 which bp, and were 226 subjected bp respectively, to conventional as shown PCR in Figures reaction 2 perihepatitis, congestion of lung and pericarditis of heart (5 andwere 3. positive and gave clear bands at the specific amplicon and 6). The bacteriological investigation for 120swabs sample broiler chicks in this study are reported in the table 6. The from broilers suffering from bumblefoot lesions revealed The hematological findings of naturally infected that S. aureus encountered in this study. The overall percentage of S. RBCs count, Hb concentration and PCV in the diseased birds hematological parameters revealed a significant decrease in aureus was the most frequent bacterialS. aureus isolate was indicated anemia as compared with the control group. Also, from legin lesions this study of bumlefoot was 45.8% diseased (55/120). cases. In addition, and monocyte in diseased groups as compared with control itisolated was isolated in pure in formmixed in form a percentage with other of bacterial 18.18%(10/55) species a significant increase in total leucocytic counts, lymphocyte from the bumble foot diseased cases and also from lung and increases of RBCs, Hb concentration and PCV levels were groups. In the levofloxacin treated group, significant

Figure 2: Showed Agarose gel electrophoresis showing positive amplification of the product 570bp fragment of Coa gene of S.aureus performed with specific primer. L: 100–1500bp DNA ladder; Pos: positive control of S.aureus; lanes1-10: 1, 2, 3, 5, 6, 9&10 +ve results of S.aureus; lane 4, 7and 8 –ve result; Neg: negative control.

Vetry Sci Rech 2019 04 www.innovationinfo.org

Figure 3: Showed Agarose gel electrophoresis showing positive amplification of the product 226bp fragment of spa gene of S.aureus performed with specific primer. Lane L: DNA molecular size marker (100-600bp), lanes 1-10:1, 3, 5, 6, 7, 8, 9&10 +ve results of S.aureus; lane 2and 4 –ve result; lane (Pos): positive control and lane (Neg): negative control.

Table 6: Hematological parameters in apparently healthy naturally infected and treated broiler chicks in farms. N=20. serum was observed in diseased birds when compared with Groups/ Apparently Diseased broiler Treated chicks by present study, the significant increase ofIL-6 and TNF-αin Parameters healthy chicks levofloxacin 6 a b a RBCs (10 /µl) 6.5±0.15 3.17±0.2 7.1±0.42 treated birds. The levofloxacin treated group revealed an a b a HB (Gm %) 11.5±0.3 7.1±0.22 10.82±0.43 Discussionimprovement in the levels of IL-6 and TNF-αin serum. PCV (%) 35.0±0.43a 21.3±0.66b 33.1±0.6a WBCs (103/ µl) 9.95±0.17b 13.55±0.2a 9.86±0.22b Lameness or leg weakness is symptomatic words Heterophile describing a condition resulting from several causes either 5.50±0.32a 3.50±0.22b 5.20±0.28a (103/ µl) bacterial or viral. It is adversely affected by the poultry Lymphocyte performance and may increase morbidity and mortality 4.00±0.15b 8.50±0.13a 4.20±0.33b (103/ µl) Monocyte (103/ 0.35±0.07a 1.40±0.06a 0.32±0.05a chondronecrosis with osteomyelitis were considered the μl) mostlevels common in broiler causes flocks. of lameness Purulent in commercial arthritis, bacterialbroilers Eosinophile 0.10±0.11a 0.15±0.06a 0.14±0.04a (103/ μl) [33]. In the present study, the observed clinical signs of Table 7: Some biochemical and immunological parameters in apparently healthy naturally infected and treated broiler chicks in farms. N=20. to stand, foot bad dermatitis, reluctance to move, gradual Treated broiler Apparently Diseased broiler diseased birds were swollen hock joint, keel bone, unable Group /Parameters chicks by healthy chick with [34,35]. Postmortem examination of naturally infected levofloxacin emaciation and finally death. The same signs were recorded ALT (u/l) 9.50±0.08 b 13.40±0.11 a 9.23±0.09 b AST (u/l) 117.5±0.3 b 152.3±0.4 a 111.33±0.32 b Uric acid (mg/dl) 8.0±0.4 b 11.40±0.32 a 8.7±0.5 b ofchickens internal showing organs. swelling These results of joints agreed filled with inflammatoryrevealed that Creatinine (mg/dl) 1.50±0.02 b 2.00±0.01 a 1.62±0.06 b exudates, arthritis of the hock, stifle, as well as congestion T. protein (gm/dl) 0.01b 8.35±0.12c 10.36±0.09a Albumin (gm/dl) 5.93±0.05a 3.50±0.01b 6.31±0.06a postmortemarthritis of the examination hock, stifle, coxofemoralof naturally joints infected and vertebralchickens Globulin (gm/dl) 3.11±0.04 b 4.85±0.05 a 4.05±0.2 b showedosteomylitis. swelling Youssef and andnecrosis Dalia, of (2012) the comb, [35] necrosis reported and the χ γλοβυλιν (γµ/δλ) 1.92±0.01 b 2.50 ±0.01 a 2.00±0.03 b congestion of liver, spleen, kidney and lungs. Other birds had βγλοβυλιν (γµ/δλ) 0.19±0.14a 0.20±0.19 a 0.22±0.15a α γλοβυλιν (γµ/δλ) 1.50±0.03 b 2.20±0.05 a 1.60±0.2 b Staphylococcus is normal inhabitants of the skin and IL-6 (pg/ml) 122.82±2.5b 139.2±2.2a 126.4±3.2b affected swollen joint filled with inflammatory exudates. TNF-α (pg/ml) 48.27±0.56b 66.3±0.84a 53.45±0.92b mucous membranes and common organisms in environments where poultry were hatched, reared or processed. Most Values with superscripts a, b and c differ significantly (P<0.05) in a row Staphylococcus shown in compared with the diseased group and they were but when a breakdown in the natural defense mechanism closely related to an apparently healthy group. of host occurs, speciesa pathogenic are considered infection will to beraise normal leading flora, to In addition, the results of some biochemical and decrease in the rates of weight gain, egg production and immunological parameters in naturally infected and treated great economic losses in broiler farms [35]. According to the bacterial investigation results of increase in (AST & ALT), uric acid, creatinine, an elevation bumblefoot or arthritic broiler and or respiratory diseased broiler chicks are revealed in the table 7. A significant in the alpha and gamma globulins. Hypoproteinemia, cases, S. aureus encountered in the recent study (45.8%) among 120 diseased in diseased group were recorded as compared with examined cases either was in the pure most or mixed frequent form. bacterial Similar results isolate hypoalbuminemia and significant increase in total globulin were achieved with [36,37] who detected the prevalence improvement in the levels of total protein, albumin, gamma, of septic arthritis caused by S. aureus within the range of andcontrol alpha group. globulin The as levofloxacin compared to treated the diseased group group. revealed In the an 50.98% collected from different broiler chicken farms with

Vetry Sci Rech 2019 05 www.innovationinfo.org symptoms of arthritis. Meanwhile, higher prevalence ratio It is the most common avian pathogens which produce of S. aureus a variety of toxins and enzymes that may contribute to (Feizi et al., 2012) [38] stated that a high prevalence ratio pathogenicity [50,51]. John Barnes (1997) [52] stated (85.71%) ofwas septic recorded arthritis up due to 81%to S. fromaureus hock in broilersjoint. Also, in that Pseudomonas infection might occur due to either skin S. aureus (36.6%) of contaminated syringe needles during egg dipping or egg Iran. Differently, lower isolation rate of inoculation,wounds or inadequate the use of contaminated hygiene like (contaminated premises from vaccines, infected arthritic broiler joints in Sharkia GovernorateS. aureus [39].infection in The most frequent sites of flock).The occurrence of antimicrobial resistance in badpoultry management, are bones, dietary tendons, and sheaths traumatic and factors joints, within especially the staphylococci of poultry origin has increased over time in ’stibiotarsal enclosure, and stifle including joints improper [36]. Arthritis perching, was poor claimed hygiene, to piercing on the bottom of the foot and leg fractures [40]. in poultry husbandry. Indeed, antimicrobial agents have been Moreover, pneumonia caused by S. aureus represents a administeredpoultry farms fordue many to the years, frequent not onlyuse of to antimicrobial control and prevent agents disease, but also for growth promotion and improved feed patients affected by immunosuppressive therapy [41]. serious complication in individuals with cystic fibrosis and high resistance of most antibiotics (trimethoprim, oxaciliin, In addition, mixed cultures of S. aureus with other conversion efficiency. The results indicated moderate toS. bacterial species were also isolated. The results indicated aureus, E.coli,Pr. Mirabilis and Ps.aeruginosa isolated strains that 32 isolates were mixed with E.coli , 8 isolates were wereenerofloxacin, recorded. tetracyclineHowever, and erythromycin) against mixed with Pr. Mirabilis Pseudomonas sensitivity rate against all isolated bacteria followed by as shown in table 3. In the same way [42] recovered that levofloxacin developed the highest four positive S. aureus isolatesand fivemixedout of sixteen with isolates in pure cultures, 10 isolates were mixed with E.coli and 2 isolates gentamycin, ciprofloxacin, and amoxicillin with varying were mixed with Proteus mirabilis. Similar results of mixed degreesGram-positive of sensitivity bacteria as shownincluding in the S. tableaureus 5. Levofloxacin[53]. Also, infection of S. aureus with E.coli (28.6%), or Salmonella is(Rohner a fluoroquinolone et al., 1992) with [54] improvedshowed that activity this group in vitro exhibited against (1.3%), or Pseudomonas (1.95%) and Pasteurella multocida an additive effect in combination with standard anti- (1.95%)were isolated from arthritic broiler cases [43]. In staphylococcal therapy in severe S. aureus infections in some addition, (Pleydell et al., 2007 and Lutfulkabir, 2010) [44,45] stated that the recovered isolates of Staphylococcus spp., S. from the infected bone of broilers; were mixed with other experimentalaureus [55] and studies. demonstrated Levofloxacin, good the activity isomer ofin theforeign race bacteria like E.coli, Mycobacterium avium, Salmonella spp., matebody ofloxacin,experimental has modelsa high level [56]. of Based in vitro on activitypharmacokinetic against and Enterococcus. Serotyping of the examined E.coli parameters, when levofloxacin of a dose of (10 mg/kg b.w); the most prevalent serogroups of E.coli isolates were O78 maintains effective plasma concentrations with bacterial isolates clarified that was given intramuscularly/orally every 24 h in turkeys, it followed by O55, O158 then O86A, O1 and O11 as shown in the infections [31]. table 3. Nearly similar results were detected by [46] reported The virulence of S.aureus is complex. It depends on that serotype O was the most common (28.2%) from 78 an array of virulence genes which are clustered into 2 diseased broiler chicken. Also, [43] Labdah et al., (2015) categories: cell surface associated (adherence) and secreted isolates into 6 different serotypes: E.coli (exotoxins) genes [57]. Coa gene (or coagulase O (30), O (14), O (16), O (6) and O (4). 78 125 55 166 146 S. aureus strains. classified seventy As regards to Congo red binding activity as a pathogencity It is an extracellular protein that has traditionally) isbeen a major used factor of E.coli , todeterminant differentiate factor coagulase for the positiveidentification Staphylococcus of from the while the other serotypes did not bind Congo red dye up less virulent coagulase-negative Staphylococcus [58]. to 48 hours post19/32 inoculation. (59.4%) These isolates results gave werepositive similarly result to [47] who observed that 20 isolates of 56 (35.7%) were in all examined S. aureus strains was cleared at 570 bp. It Congo red positive and 36 isolates (64.3%) were negative wasIn recovered this study, in the 70% amplified of the examined product ofisolates the coagulase in this study. gene from broiler chickens in Egypt. Also, [48] showed that 32 out In addition, Spa gene which encodes for protein A; is mostly of 53 E.coli isolates (60.4%) were Congo red positive. used for typing of S. aureus [59]. Protein A is an important exo- In this study, as shown in table 3, a lower isolation rate protein virulence factor which enables the microbe to evade (6.67%) of Pr. Mirabilis in mixed culture with S. aureus was recorded. In the same manner, [49,43] Hassan et al., 2012 covalently anchored to the peptidoglycan of S. aureus. About host immune responses. Its molecular weight of 42KD. It is mentioned that Pr. mirabilis was isolated from 18 months 90% of protein A is found in the cell wall and the remaining old hens with Bumblefoot and it was able to re-induce the 10% is free in the cytoplasm of bacteria. In MRSA strains of disease experimentally when inoculated into the footpad S. aureus, protein A is unable to adhere to the cell wall and of chickens. In the same table, a mixed infection with P. therefore is released into the media (secretary protein) [60]. In this study, it was found in 80% of the examined isolates. samples. This result nearly similar to (Hebat-allah, 2004) Vintov et al. (2003) [61] cleared that the polymorphic coa recordedaeruginosa Ps. was aeruginosa isolated infrom 4.17% 3.3% (5/120) of examined of all examined broilers. and spa genes could be used to investigate the diversity of S. aureus.

Vetry Sci Rech 2019 06 www.innovationinfo.org The obtained results of hematological parameters Our results are completely agreed with [11,71,72,77] who reported highest creatinine and urea levels in case of the concentration and PCV in the diseased birds indicated anemiarevealed that a was significant correlated decrease with the severity in RBCs of infection count, Hb of increased in respiratory affection birds. This increase bacterial organisms. These results were in accordance with mightrenal disease.be attributed Uric acid to the and increased creatinine protein were significantlycatabolism, febrile respiratory diseases, impaired cardiac function and lymphocyte, and monocyte between infected and control groups[62,11]. was Also, recorded. a significant Similarly, increase Sakiniene in total leucocyticet al. (1999) counts, [63] Interleukin-6 is one of the most pleiotropic interleukins reported that S. aureus infection could lead to leukocytosis decreased renal blood flow [78]. and [64] Lucke et al. investigators reported that IL-6 could activate osteoclasts in total leucocytic count in infected animals. In the released at sites of injury or infection [79]. Some (2003) recorded a significant increase process [80]. Increased serum IL-6 levels had been observed Hb concentration and PCV levels were shown in compared throughoutwhich increase the thecourse damage of S. of aureus joints duringarthritis the in arthriticseveral withlevofloxacin the diseased treated group group, and they significant were closely increases related of RBCs,to an animal models. Interleukin-6 is produced by many different cell types and acts on B-lymphocytes, T-lymphocytes, decrease in leukocyte in the treated group when compared hepatocytes, hematopoietic progenitor cells, and cells of withapparently the diseasedhealthy group. group. Our These results results indicated indicated a significant the the central nervous system [79]. In the present study, the Lee et al., (2017) [66] showed that the administration of in diseased birds when compared with treated birds, which levofloxacin antibacterial effect on microorganisms [12,65].E.coli. wassignificant similar increase to the reportsofIL-6 and by TNF-αin(Zhou et serum al., 2007) was observed[79] in a 5mg/kgS. aureus of levofloxacin secretes proteinsseems to whichbe effective inhibit in killingcomplement model of staphylococcal arthritis. However, the S. aureus- activation, lyes neutrophils, neutralizes antimicrobial peptides and reduces the effectiveness of neutrophil. It could levels of IL-6 in serum, as compared with the diseased survive in phagosomes, express polysaccharides and proteins birds.infected Zhou birds et al treated., (2007) with [79] levofloxacin showed that showed IL-6 activities reduced which inhibit opsonization by antibody and complement, and concentrations were reduced (P < 0.05) in the serum of and it’s cell resistant to lysozyme. Furthermore, S. aureus has several types of super antigen that corrupt the normal S. aureus. humoral immune response, resulting in immunosuppression infected broilers treated with levofloxacin compared with [67]. A characteristic manifestation of S. aureus-caused birds injected only with Tumor necrosis factor-α (TNF--α) is the major macrophage- characterized by a rapid and excessive recruitment of derived cytokines present in the rheumatoid joint and both neutrophilspneumonia to is the the site intense of infection host [68]. inflammatory response ofinduce a molecule the synthesis called and “cachectin”, secretion whichfrom synovial causes wastingfibroblasts in The increase in serum AST and ALT levels in this work of matrix-degrading proteases. The purification and cloning could be due to liver damage produced by the infected bacteria. Campbell and Coles, (1986) [69] mentioned that consideredchronic diseases, to be mediatedwas subsequently through foundthe interactions to be identical among to the increased of the activity of AST had been associated cytokines,TNF-α. In general,including the the systematic activation inflammatory of tumor necrosis response factor is with hepatocellular damage in birds. Also, they reported (TNF) and the corresponding receptors and neuroendocrine elevation of ALT in birds which were infected with bacteria. increase in (AST & ALT) in chicken infected with E.coli . The bypathways lymphocytes, (Woodcock monocytes and and Morganti-Kossmann, macrophages, but a number 2013). Our result agreed with [70,71] who observed a significant TNF-α is produced primarily by macrophages, a lesser extent work could be due to liver and kidney damage which could keratinocytes and smooth muscle cells, also produce TNF. significant change in total protein and albumin in the present of non-immune cell types, including fibroblasts, neurons, previously mentioned by [69,71-73]. immune response and is an acute-phase protein that initiates be associated with bacterial infection. Similar findings were aTNF-α cascade acts ofas cytokines.a key intermediary Furthermore, in the high local levelsinflammatory of TNF Bacterial toxins increase the capillary permeability result in increased vascular permeability, thereby recruiting and permitted the escape of plasma proteins into tissue resulting in hypoproteinemia [74]. An elevation in the alpha infection ( Esposito and Cuzzocrea, 2009 and Woodcock and and gamma globulins usually indicated an activation of the macrophages and neutrophils to the site of injury and/or diseases [75] and could be associated with bacterial Morganti-Kossmann, 2013). septicemiaimmune system or chronic which isinfection due to infection[74-76]. orAn inflammatory increase in family, possesses excellent potent activity against a beta and gamma globulin in chickens were recorded in wideLevofloxacin microbial spectrum. drug, a member Hu et al of., (2002) the fluoroquinolone [65] showed an experimentally infected chickens with E.coli and S. aureus staphylococcus is especially when was administered in improvement in the levels of total protein, albumin, gamma, thethat drinking Levofloxacin water. was It had effective also been for demonstrated the control ofthat avian the and alpha [76]. globulin The levofloxacinas compared treatedto the diseased group revealedgroup. an levels of tumor necrosis factor-α (TNF-α) and IL-6 in serum were associated with the severity of the infectious process. The increase in uric acid and creatinine could be due to Additionally, IL-1β and TNF-α could induce chondrocytes the effect of the microorganisms and its toxins on the kidneys.

Vetry Sci Rech 2019 07 www.innovationinfo.org and synovial macrophages to release a high production of 1st Published. 18. Ewing W H (1986) Enterobacteriaceae. 4th Ed.536. (Edwards’s) Ewing’s Identification of IL-6locally in bone, which consequently increases damage to 19. Congo red dye agar jointsConclusion during the arthritic process [79]. test as an indicator test for detection of invasive bovine Escherichia coli. VeterinarskiarchivSharma KK, Soni SS, 76 Sunil (4): Meharchandani363-366. S (2006) Staphylococcal infections in poultry are high especially 20. Antibiotic susceptibility in Bumblefoot disease or arthritis disease. Hence, due to its testing by a standardized single disc method. Am J Clin Pathol 45: 493- epidemiological importance, the measures must be taken 496.Bauer AW, Kirby WM, Sherris JC, Turck M (1966) 21. Clinical and Laboratory Standards Institute (CLSI) (2011) Performance from poultry to humans. The hematological, biochemical in the field to minimize the zoonotic disease transmissible informational supplement, CLSI document M100-S21; Wayne, PA,USA.; 31:standards 42-46. for antimicrobial susceptibility testing; twenty-first 22. Jain N C (2000) Schalm’s veterinary hematology.8th . Ed. Lea and succeededand immunological in the treatment analysis of the (IL-6 disease and TNF)in the couldbroiler reflect farm. Febiger, Philadelphia, U.S.A. the pathogenesis of the microbe. Levofloxacin treatment References 23. Reitman S, Frankel S (1957) A colorimetric method for determination of AST and ALT. Am. Am J Clin Pathol 28: 56-63. 1. Hester P Y (1994) The role of environment and management on leg abnormalities in meat-type fowl. Poultry Sci 73: 904-915. 24. Henry R J (1979) Colorimetric methods for determination of serum creatinine 2. Trauma Medicine. Avian Medicine: Principles and and Row. Application. Wingers Publishing Inc. Florida. . Clinical Chemistry, Principles and techniques, 2nd ed. Harper Degernes L A (1994) 25. Caraway W T (1963) Uric acid. Standard Methods of Clin. Chem 4: 239- 3. Enterotoxigenicity of 247. Staphylococcus aureus strains isolated from chickens. J Food Prot 43: 683-685.Shiozawa K, Kato E, Shimizu A (1980) 26. Henry R J (1974) Colorimetric method for determination of serum total protein. Clinical Chemistry, Harper and Row Publishers, New York. 4. Antimicrobial susceptibilities of Staphylococcus aureus isolated from commercial 27. Colorimetric determination of serum albumin. Clin broilersWhite DG, in Ayersnortheastern S, Maurer Georgia JJ, Thayer SG, Hofacre C (2003) Chem Acta: 400-403. Doumas B (1971) 5. McNamee PT, Smyth JA (2000) Bacterial chondronecrosis with 28. . In: . Avian Dis 47: 203-210. osteomylitis (femoral head necrosis) of broiler chickens: a review. Avian Standard Methods of Clinical chemistry. 7, ed. G. R. Cooper. New York Pathology 29: 477-495. AcademicDoumas BT, press. Bigs H G (1972) Determination of serum globulin 6. Chansiripornchai V R (2009) 29. Cleavage of structural proteins during the assembly vaccine in Broilers against avian E. coli serotype O78 Infection. Thai J Vet of the head of bacteriophage T4. Nature 227: 680-685. Laemmli UK (1970) Med 39: 337-342. The Efficacy of Escherichia coli Aro A-live 30. A 7. Histopathology of purulent arthritis . Biol Psychiatry 67: 446- of chickens. Jap J.Vet. Sci 38: 451-459. 457.Dowlati Y, Herrmann N, Swardfager W, Liu H, Sham L, et al (2010) Itakura C, Kurisu K, Goto M (1976) meta-analysis of cytokines in major depression 8. Glisson JR (1998) Bacterial respiratory diseases of poultry. Poult. Sci77: 31. Bioavailability and 1139-1142. intramuscularAboubakr M1, and Uney oral administration K, Elmas M in turkeys (2014) . Br Poult Sci 55: 115- 9. Molecular survey of 119.pharmacokinetic profile of levofloxacin following intravenous, respiratoryRoussan DA, diseases R Haddad,G in Jordan Khawaldeh. Poult. Sci 87: (2008) 444-448. 32. Ryan B F, Joiner B L (2005) Minitab Handbook: update for release 14. avian respiratory pathogens in commercial broiler chicken flocks with 6th edition. 10. Garmyn A, A Martel, R Froyman, C Ludwig, H Nauwynck, et al (2009) 33. Clinical and morphological investigations on the course of respiratory disease caused by avian metapneumovirus and prevalence of lameness, associated with femoral head necroses in ornithonThe effect bacterium of reduced rhinotracheale treatment time. Poult and Scidosage 88: 2315-2323. of enrofloxacin on the broilerDinev Ichickens (2009). British Poultry Science 3: 284-290. 11. Mona S Zaki, Olfat, Fawzy ,Osfor M H (2012) Effect of E. coli O: 157 34. Bakheet A A (2011) Bacterial Aspects of Arthritis in Broiler Chickens. on Baladi Broiler Chicken and some Biochemical studies. Life Science Zagazige vet J 39: 22-30. Journal 9: 91-94. 35. Methicillin Resistant Staphylococcus 12. A review of its antibacterial aureus (MRSA) associated with arthritis in broiler farms in Ismailia Youssef A I, Dalia M Hamed (2012) . SCVMJ 17: 309- Davis R, Bryson HM (994) Levofloxacin. 321. 13. activity, pharmacokinetics and therapeutic efficacy. Drugs 47: 677-700. province, Egypt and its zoonotic potential significance . Hum Exp 36. Adayel S A (2005) Incidence of Staphylococcus aureus causing Arthritis Toxicol18:Takizawa T,392-399. Hashimoto K, Minami T, Yamashita S, Owen K (1999) of broiler dreeders. 4th Int. Sci. Conf. Mansoura. The comparative arthropathy of fluoroquinolones in dogs 14. 37. Rasheed B Y (2011) arthritis in chickens Isolation and identification of bacteria causing birdsPatel . JProceedings H, R D Varia, of the U D8th Patel, Annual S KConference Bhavsar, Aof MIndian Thaker Society (2008) of 38. Feizi A, Nazeri M, Pilevar A (2012) Isolation of Staphylococcus spp. VeterinaryPharmacokinetics, Pharmacology bioavailability and Toxicology and safety and of levofloxacinNational Symposium in broiler . Iraqi Journal of Veterinary Sciences 25: 93-95. Prevalence and antimicrobial susceptibility. African J Microbiol Res 6: 5819-5823.Genera from broiler breeder flocks in East AzerbaijanProvince of Iran: on Challenges, Scientific Validation and IPR Protection of Indigenous 39. Isolation of some bacterial agents associated with MathuraMedicinal India. Plants Based ITK and Emerging Risks to Wildlife Due to Drugs lesions in chickens in Sharkia Province of Egypt, Zag. Vet J 33: 100-107. and Toxicants and Ameliorative Measures, November 6-8, DUVASU, 15. Omayma KI (2005) 40. The Australian Registry of Wildlife Health (2005) . Indian veterinary Urban Wildlife: Birds. Part 2. Accessed on 14 Jan 2019. Varia R.D, Patel jH, Vihol UD, U D Patel, SK Bhavsar, et al (2012) Common Diseases of Evaluation of levofloxacin safety in broiler chickens 41. Antibiotic 16. Beutin L, Montenegro M A, Orskov I, Orskov F, Prada J, Zimmermann journal 89: 54-56. S, Stephan R (1989) Close association of verotoxin (Shiga-like toxin) Conway SP1, Brownlee. Am J Respir KG, Denton Med 2: 321-32. M, Peckham DG (2003) production with enterohemolysin production in strains of Escherichia treatment of multidrug-resistant organisms in cystic fibrosis organisms 42. Hassan A H, Hussein S A, AbdulAhad E A (2012) Pathological and coli. J. Clin. Microbiol 27: 2559-2564. in cystic fibrosis bacteriological study of Bumblefoot cases in Sulaimaniyah province. Al- 17. Anbar J Vet. Sci.5. . Blockwell Science Ltd. Quinn PJ, Markery B K, Carter M E, Donnelly WJ, Leonard F C (2002) Veterinary Microbiology and Microbial Diseases Vetry Sci Rech 2019 08 www.innovationinfo.org 43. Lebdah M A, Fatma M Youssef, Elwan E A (2015) Bacterial Leg Infections 61. Vintov J, Aarestrup FM, Zinn CE, Olsen JE (2003) Association between in Broiler Chickens. Zagazig Veterinary Journal 3: 179-188. phage types and antimicrobial resistance among bovine Staphylococcus 44. aureus from 10 countries. Veterinary Microbiology 95: 133-147. Sources of variation in the ampicillin-resistant Escherichia coli 62. Jain N C (1986) Schalm’s Veterinary Hematology. 4thEdn, Lea and concentrationPleydell EJ1, Brown in the PE,feces Woodward of organic MJ, broiler Davies chickens RH, French. Appl NP Environ (2007) Febiger, Philadelphia. Microbiol 73: 203-210. 63. Sakiniene E, Bremell T, Tarkowski A (1999) Complement depletion 45. Lutfulkabir S M (2010) Avian colibacillosis and salmonellosis: A closer aggravates Staphylococcus aureus septicemia and septic arthritis. Clin look atepidemiology, pathogenesis, diagnosis, control and public health Exp Immunol 115: 95-102. concerns. Int J Environ Res Public Health 7: 89-114. 64. Lucke M, Schmidmaier G, Sadoni S, Wildemann B, Schiller R (2003) A 46. Clinical, new model of implant- related osteomyelitis in rats. J Biomed Mater Res Pathological and Bacteriological Investigations on Air Sacculitis in B Appl Biomater 67: 593-602. Fatma M Yousseff, Mona A Ahmed, Dalia H Mansour (2008) Chickens in Ismailia Province (Egypt). Journal of Agricultural and 65. Hu, G Z, L J, Xu, L Yuan, et al (2002) . Veterinary Sciences 1: 71-79. Chin J Huazhong Agric Univ. Pharmacokinetical of levofloxacine 47. Amer M M, Mekky HM, Amer A M, Fedawy H S (2018) Antimicrobial 66. Ex vivo resistance genes in pathogenic Escherichia coli isolated from diseased broiler chickens in Egypt and their relationship with the phenotypic Lee HK, DeVito V, Vercelli C, Tramuta C, Nebbia P, et al (2017) resistance characteristics. Vet World 1: 1082-1088. inantibacterial broilers. Res activity Vet Sci of112: levofloxacin 26-33. against Escherichia coli and its pharmacokinetic profile following intravenous and oral administrations 48. Abdel Ameer H Zahid, Muna Turkey Mossa AL-Mossawei, Aaisha B 67. Foster TJ (2005) Immune evasion by Staphylococci. Nat Rev Microbiol Mahmood (2016) In vitro and In vivo Pathogenicity tests of Local 3: 948-58. Isolates APEC from Naturally Infected Broiler in Baghdad. Int. J. Adv. Res. Biol. Sci 3: 89-100. 68. Bubeck Wardenburg J, Patel RJ, Schneewind O (2007) Surface proteins 49. Pathogenicity of Proteus mirabilis in aureus pneumonia. Infect Immun 75: 1040-1044. Bumblefoot in chickens and exotoxins are required for the pathogenesis of Staphylococcus Ahmed H E, El Amin E D M (2008) 69. Campbell T, Coles E (1986) Avian clinical Pathology, ‘in “Veterinary 50. . The Sudan j. Vet. Res 23: 91-95. Clinical pathology” .4th Ed., W.B Saunders Company. Philadelphia. tierärztliche Wochenschrift: wissenschaftliche Zeitschrift für die London and Toronto. VeterinärmedizinHinz K H, Ryll M, HeffelsTieraerztlwochenschr Redmann U, Po ppel 99: 75-78. M (1992) Deutsche 70. Omaima A R (1987) Liver function tests in relation to some bacterial and 51. . Dtsch viral diseases in chickens. MVSc, of Biochemistry- Faculty of Vet. Med. Zagazig University (Benha branch). fromLin MY, sick Cheng or dead MC, chickensHuang KJ, Tsai WC (1993) Classification, Pathogenicity, and drug susceptibility of hemolytic gram–negative bacteria isolated 71. Mona A Ahmed, Fatma M Youssef, Abdel Rahman AG (2013) 52. John Barnes H (1997) Other bacterial disease: Pseudomonas . Avian Dis 37: 6-9. of Poultry.10th edition: 291-292. chicken by using multiplex PCR. Egypt J Vet Sci 44: 21-36. . In Diseases Differentiation between E. colis strains causing diarrhea in broiler 53. In vitro and 72. Pai CH, Gordon R, Sims HV, Bryan LE (1984) Sporadic cases of hemorrhagic colitis associated with Escherichia coli 0157: H7. Clinical. Fu KP1, Lafredo SC, Foleno B, Isaacson DM, Barrett JF (1992) . Antimicrob Agents Chemother 36: 860-866. Epidemiologic and bacteriologic features. Ann Intern Med 101: 738-742. in vivo antibacterial activities of levofloxacin (l‐ofloxacin), an optically 54. Rohner P, Herter C, Auckenthaler R, Pechere J C, Waldvogel F A, et al active ofloxacin 73. Infections with Escherichia (1992) coli 0157: H7 an emerging gastrrointestinal pathogen. Results of a one- resistant Staphylococcus species. Antimicrob Agents Chemother 33: year,Ostroff prospective, SM, Kobayashi population-based JM, Lewis JH study(1989). JAMA 262: 355-359. 2037-2041.Synergistic effect of quinolones and oxacillin on methicillin‐ 74. Coles E (1986) “Veterinary Clinical Pathology”. 4thed. W.B. Sanders 55. company, Philadelphia, London, Toronto, Mexico City, Sydney, Tokyo, treatment of bacterial infections in the United States Croom2802. KF, Goa KL (2003) Levofloxacin: a review of its use in the 75. Butler E J (1983) “Plasma proteins.” In: Physiology and Biochemistry of . Drugs 63: 2769- Hong Kong. 56. . B. M. Academic Press.London. 76. Fatma M Yousseff (2005) Clinicopathological studies on the effect of infectionMurillo O, by Doménech methicillin-susceptible A, Garcia A, Staphylococcus Tubau F, Cabellos aureus C, . et Antimicrob. al (2006) the Domestic Fowl AgentsEfficacy Chemother of high doses 50: 4011-4017. of levofloxacin in experimental foreign-body . 57. The role of virulence determinants in jojoba seeds as antibacterial agent and immunostimulant in chickens 77. community-associated MRSA pathogenesis. Trends in Microbiology 16: Department of Clinical Pathology. Fac. Vet. Med., Suez Canal University. 361-369.Diep BA1, Otto M (2008) (1987) The pathogenesis of hemorrhagic colitis caused by Escherichia coliTzipori 0157: S, H7 Wachsmuth in gnotobiotic IK, Chapmanpiglets C, Birden R, Brittingham J,et al 58. Ahlam A. Gharib, M.A. Adel Attia and M.M. Bendary (2013) 78. Abdalla O E, Emam E E (2005) of the Coa Gene in Staphylococcus aureus from different Sources by . J.Infect Dis 154: 712-716. Polymerase Chain Reaction. International Journal of MicrobiologicalDetection Evaluation of mabofloxacin and Research 4: 37-42. Pasteurella multocida in lambs. 4th Int. Sci. Conf. Mansoura. isoflupredone acetate as therapy of pneumonia associated with 59. 79. Role of interleukin-6 Alang, Hamid Vaez, et al. (2010) in the pathogenesis of an avian model of Staphylococcus aureus arthritis. Zhou Q1, Gu CQ, Hu XY, Wang DH, Li XM, et al (2007) StrainsFatemeh of Shakeri, Staphylococcus Abolfath aureus Shojai, in Masoud North of Golalipour, Iran. International Somaye JournalRahimi Poult Sci 86: 1245-1250. of Microbiology 2010: 5. Spa Diversity among MRSA and MSSA 80. Interleukin-6 attenuates 60. Movitz J (1974) A study on the biosynthesis of protein A in Staphylococcus agonist-mediated calcium mobilization in murine osteoblastic cells. J Green J, Schotland S, Sella Z, Kleeman CR (1994) aureus. Eur J Biochem 48: 131-136. Clin Invest 93: 2340-2350.

Citation: Youssef FM, Soliman AA, Ibrahim GA, Saleh HA (2019) Advanced Bacteriological Studies on Bumblefoot Infections in Broiler Chicken with Some Clinicopathological Alteration. Vetry Sci Rech Vol: 1, Issu: 1 (01-09).

Vetry Sci Rech 2019 09