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Bovine Respiratory Disease (BRD) complex in buffaloes--a study of associated bacterial isolates and their antibiograms.


Diseases impacting bovine respiratory tract have been grouped into an overall category known as Bovine Respiratory Disease (BRD) complex. This includes shipping fever syndrome, mucosal disease, enzootic calf pneumonia, acute respiratory distress syndrome, hemorrhagic syndrome and atypical interstitial pneumonia (Apley, 2006). BRD complex is a result of complex interaction of bacterial and viral agents, environmental conditions, managemental factors and animal health (McVey, 2009; Nickell and White, 2010). Though viral agents can induce clinical signs without bacterial infection, naturally occurring BRD complex generally involves concurrent infection with bacteria (Hogdson et al., 2005; Taylor, 2010). These bacteria are considered commensals of upper respiratory tract and nasopharynx, however stress and viral infection are predisposing factors for bacterial infection in lower respiratory tract (Apley, 2006; Confer, 2009; Griffin et al., 2010). BRD complex is a major health problem occurring worldwide in both dairy and feedlot cattle (Edwards, 2010) responsible for high morbidity and mortality and has been reported to cause heavy economic losses in terms of drug and Veterinary costs, extra labour and production losses (Van der Fels-Klerx et al., 2001; Gagea et al., 2006). The disease is estimated to cost $800 million-$1 billion annually in US (USDA/Agriculture Research Service, 2011), but there is no report regarding the economic loss in India due to BRD complex. Antimicrobial therapy is the most effective method for prevention and treatment of BRD complex (Gibbs, 2001). Therefore, the aim of this study was to identify the bacterial isolates from nasal swabs of BRD complex infected buffaloes and determine their antibiogram profile.

Materials and Methods

History, Clinical observations and Diagnosis

The present study was conducted on 24 clinical cases of BRD complex in buffaloes admitted with complaint of serous to mucopurulent nasal and ocular discharge, moist cough, rapid and shallow respiration, crusty muzzle, prolonged inappetance, high fever and depression. These cases were brought from different places of Haryana and border areas of Rajasthan state. The diagnosis was based on clinical signs, haematological findings (leuckocytosis with neutrophilia) and confirmed by cultural and radiological examination.

Collection of nasal swab samples and bacteriological examination

Nasal swabs from affected animals were collected aseptically and brought to Central Laboratory, Veterinary college for cultural examination and determination of antimicrobial sensitivity pattern. For bacteriological examination, nasal swab samples were inoculated and streaked on blood agar and MacConkey's lactose agar plates and incubated at 37[degree]C for 18-24 hours. Primary identification of bacterial isolates was undertaken on basis of colony morphology, Gram's staining, catalase and oxidase tests (Quinn et al., 2004). Bacteria were further characterized upto species level on basis of biochemical tests. Antimicrobial susceptibility testing was performed on all isolates employing standard disk diffusion method (Bauer et al., 1966).

Results and Discussion

Different bacteria isolated in order of frequency from a total of 24 nasal swab samples taken from buffaloes affected with BRD complex were Staphylococcus aureus (37.50%), Streptococcus pyogenes (29.17%), E. coli (29.17%), Diplococcus spp. (8.33%), Pasteurella multocida (4.17%) and Klebsiella pneumoniae (4.17%) including 3 cases of mixed infection in combinations of Staphylococcus aureus + Streptococcus pyogenes, Staphylococcus aureus + Diplococcus spp. and Staphylococcus aureus + Klebsiella pneumoniae (Table 1). Our study represents Staphylococcus aureus (37.50%), Streptococcus pyogenes (29.17%) and E. coli (29.17%) as major bacterial agents responsible BRD complex in buffaloes, whereas, Mohammadi et al. (2006) isolated Staphylococcus spp., Streptococcus spp. and E. coli only from 2.31%, 3.08% and 3.84% cases in calves respectively. Most of the workers (Mohammadi et al., 2006; Portis et al., 2012; Kurcubic et al., 2013) reported Pasteurella multocida, Mannheimia haemolytica, Bacillus spp., Staphylococcus spp., Streptococcus spp., Pseudomonas spp., Proteus spp., E. coli, Histophilus somni, Aeromonas viridians and Corynebacterium bovis as different bacteria isolated in order of frequency from respiratory tract of bovine particularly cattle from different parts of world indicating Pasteurella multocida as most important causative agent responsible for BRD complex in bovines. This difference in type of bacterial pathogens isolated could be due to that the work reported by above workers has been conducted in cattle, whereas our work was in Murrah buffalo (Bubalus bubalis). Another important reason could be that Pasteurella multocida mostly causes acute disease (Haemorrhagic Septicaemia) in buffaloes and affected animals either treated and recovered in field (if treatment provided as early as possible) or died even before reaching to referral hospital. Apart from these BRD complex is a multifactorial disease and nasal microbial flora varies greatly with age, geographic location, nutrition and climate (Yimer and Asseged, 2007) and relatively poor hygienic and sanitary measures maintained for animals under Indian conditions. An interesting finding in our study was isolation of Diplococcus spp. from BRD complex affected buffaloes because usually bacteria affects human population. The possible reason could be that the farmers under Indian conditions are in close contact with their livestock population.

Various antimicrobials have been tested and used with variable efficacy for therapeutic purpose in BRD complex from time to time, which are selected by Veterinarians on basis of perceived efficacy, cost, convenience, availability, toxicity and residue profile (Songer and Post, 2005). Commonly used antimicrobial drugs are Penicillin, Enrofloxacin, Oxytetracycline, Ceftiofur, Levofloxacin, Florfenicol and Tulathromycin, Marbofloxacin and Ciprofloxacin. New infection, recrudescence of pre-existing infection, misdiagnosis and irreparable damage caused by previous disease may also influence response to treatment (Radostits et al., 2007). Keeping in view, the multifactorial nature of disease, 21 different antimicrobials were tested against different bacterial isolates from nasal swabs of buffaloes affected with BRD complex (Table 2). Results of antibiogram profile revealed Ceftiofur as the most sensitive (100%) antimicrobial against these isolates followed by Levofloxacin (87.5%), Amikacin (83.33%), Moxifloxacin (79.6%), Gentamicin (75%), Neomycin (75%), Chloramphenicol (70.83%), Cephalexin (70.83%), Enrofloxacin (66.6%), Ciprofloxacin (62.5%), Ceftriaxone (62.5%), Clindamycin (62.5%), Cefoperazone (58.33%), Cloxacillin (58.33%), Pencillin-G (58.33%), Streptomycin (50%), Tetracycline (50%), Co-trimoxazole (45.83%), Amoxyclav (41.66%), Ampicillin (37.5%) and Amoxicillin (33.33%). Even Ceftiofur (@ 2.2 mg/kg b.wt.) was found to be most effective of the tested drugs (Levofloxacin @ 4 mg/kg b.wt. and Moxifloxacin @ 5 mg/kg b.wt.) in clinical cases of BRD complex in buffaloes during our study. Ceftiofur has been reported to be most sensitive antimicrobial agent by some other researchers also (Burton et al., 1996; Salmon et al., 1996; Portis et al., 2012; Jamali et al., 2014) in cases of BRD complex and other respiratory diseases of animals. A decrease in treatment efficacy, as indicated by increased mortality or number of animals requiring re-treatment, usually encourages practitioners to submit samples for bacteriological culturing and antimicrobial susceptibility testing. Delayed diagnosis and treatment is one of the most important factor related to relapse of BRD complex in cattle (Radostits et al., 2007; Babcock et al., 2009). Thus, monitoring of antimicrobial susceptibility trends of BRD complex pathogens is an important aid to Veterinarians in selecting the most efficacious and cost-effective therapeutic agents (Mohammadi et al., 2006).


The authors are thankful to The Dean, College of Veterinary Sciences, LUVAS for the facilities provided to conduct the study.


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Praveen Kumar (1), Ashok Kumar (2), Anshu Sharma (3) and Rajendra Yadav (4)

Department of Veterinary Medicine College of Veterinary Sciences

Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS)

Hisar--125004 (Haryana)

(1.) Post Graduate Scholar

(2.) Professor

(3.) Senior Bacteriologist and Incharge, College Central Laboratory

(4.) Assistant Professor and Corresponding author. E-mail:
Table 1: Bacterial isolates from nasal swabs
of buffaloes affected with BRD complex

Bacterial isolates        No. of     Percentage
                         samples     of samples
                         positive   positive (%)
Staphylococcus aureus       9          37.50
Streptococcus pyogenes      7          29.17
E. coli                     7          29.17
Diplococcus spp.            2           8.33
Pasteurella multocida       1           4.17
Klebsiella pneumoniae       1           4.17

Table 2: Antimicrobial sensitivity pattern of
bacterial isolates from nasal swabs of BRD
complex affected buffaloes.

Antimicriobals                Percent
                          sensitivity (%)

Ceftiofur                     100.00
Levofloxacin                   87.50
Amikacin                       83.33
Moxifloxacin                   79.60
Gentamicin                     75.00
Neomycin                       75.00
Chloramphenicol                70.83
Cephalexin                     70.83
Enrofloxacin                   66.60
Ciprofloxacin                  62.50
Ceftriaxone                    62.50
Clindamycin                    62.50
Cefoperazone                   58.33
Cloxacillin                    58.33
Pencillin-G                    58.33
Streptomycin                   50.00
Tetracycline                   50.00
Co-trimoxazole                 45.83
Amoxyclav                      41.66
Ampicillin                     37.50
Amoxicillin                    33.33
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Title Annotation:Research Article
Author:Kumar, Praveen; Kumar, Ashok; Sharma, Anshu; Yadav, Rajendra
Publication:Intas Polivet
Article Type:Report
Date:Jul 1, 2015
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