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Indian Res. J. Genet. & Biotech. 5(3) : 160-164 (2013) Antibiotic sensitivity profile of the test isolates of E. coli isolated
from clinical cases of diarrhea in the kids of goat breeds
S. K. Rajput1, K. Gururaj2, D.V. Singh3 and Gitam Singh4
1, 2, 4 C. I. R. G, Makhdoom, Farah, Mathura, U.P., 3K.V.K., Nabarangpur, Odisha.
(Received : May, 2013 : Revised : June, 2013; Accepted : July, 2013) Abstract
The samples were taken using sterile cotton swab tubes, and were labeled and kept in the
refrigerator at 4°C until further processing. These samples were collected from less than one
month old Jamunapari, Barbari, Jakharana, Sirohi kids during the period from March to June
2010. Some of the commonly used antibiotics like Ampicillin, and Cloxacillin were giving high
percentage of resistance at the rate of 75 percent and 100% respectively. This is due to the
unscrupulous use of such antibiotics leading to high resistance. About 67 percent of the isolates
were resistant to Oxytetracycline, Gentamicin, and Ceftriaxone. Only antibiotic groups like
fluoroquinolones and the Chloramphenicol have fared well in effectively inhibiting the in vitro
growth of E. coli. Chloramphenicol has given the lowest percentage of resistance at zero and
the next best was Enrofloxacin at 34 percent. So, it’s a serious issue that out of the commonly
used 7 antibiotics, only two was effective, which gives clue about the emerging multi-drug
resistant E. coli strains.
Key words: antibiotic sensitivity, E. coli, kids, goat
infectious organism by looking at the color, consistency, or odor of the feces. A definitive identification requires a sample for microbiological Introduction
analysis. In livestock, diarrhea is called scours.
Goat population in the world was 464,323000 in There are many causes of diarrhea: bacterial, viral, 1980; 591,367000 in 1990; 746,689000 in 2000; parasites, and diet (Joe Rook 2007). Diarrhoeal
diseases are the top listed disease causing death 6,674,801000 human populations in 2008. Goat of kids. Colibacillosis and septicemia are generally population in India was 86,900000 in 1980; taken as the most common cause of mortality in juvenile kids in India (Vihan et al, 1990) along with 125,732000 in 2008; 39 goat breeds; 1,139,965000 other infectious organism like Cryptosporidium human population in 2008; 1,080 US$ gross national parvum, rota virus, Clostridium perfringens and income per capita (Shrestha et al., 2011).
Salmonella species. Morbidity rate (60.8%) and Diarrhea is defined as an increased frequency, Colibacillosis in kids in organized farm (Kumar et fluidity, or volume of fecal excretion. The feces may al, 2001). Outbreak of colibacillosis in established contain blood or mucous and may have foetid odour.
The color of the feces may be abnormal. However, it is not possible to definitively determine the Correspondance author's e-mail : [email protected], [email protected] by Indian Society of Genetics, Biotechnology Research and Development, 114, IInd floor Biotech Bhawan, PuspanjaliCommercial Complex, Shastripuram Road, Sikandra 282007 Antibiotic sensitivity profile of the test isolates of E. coli . Diarrhoea caused by Escherichia coli infection is and sub cultured for further characterization. To responsible for high rates of mortality in goat kids check whether the bacteria are E. coli (because which causes important economical loss in goat- there are other lactose fermenters like Klebsiella rearing. Goat kids are especially susceptible to E. and Enterobacter) the LF+ colonies were re- coli scours during the first week of life. A constant inoculated on EMB agar for presence of metallic bacteriological finding in enteric colibacillosis is the sheen, which was given only by E. coli. The intense proliferation of certain serotypes of E. coli organism was identified on the basis of culture, in the small intestine whiles the number of other morphological and biochemical characteristics bacteria in the gut remains normal (Hodgson, 1994).
(Karter, 1973). On the basis of microscopic Diseases are the major cause of economic losses examination, morphology of bacteria was noted as and inefficient production of livestock and main rod, spiral or filament. It was differentiated by Bio- constraint in development of livestock particularly of chemical characterization as per Jackie Reynolds goat industry in country. The risk of endemic diseases contributes significantly, which is Initially the Glass wares and other media are considered more obvious under Indian conditions.
sterilized and kept ready. MHA was sterilized by Materials and Methods
autoclaving at the rate 121ºC in 15 lbs for 15minutes and poured in to Petri plates and checked for E. coli was isolated from faeces of Kids suffering sterility. Bacterial Inoculums was prepared from from clinical diarrhoea. The Jamunapari, Barbari, each isolates in normal saline and matched with Jakhrana, Sirohi goat unit of CIRG Makhdoom, McFarland Standards as mentioned by Kirby & farah, Mathura was selected for collection of Bauer (1966). A lawn culture was prepared using the primary inoculums by spreading the inoculums on diarrhoea were selected and screened out. The to the Agar surface nicely by using a sterile glass samples were taken using sterile cotton swab spreader (sterilized by 70% alcohol). After 15 tubes, and were labeled and kept in the refrigerator minutes, the antibiotics discs were applied on to the at 4°C until further processing. These samples were agar surface by the applicator/ sterile forceps with collected from less than one month old Jamunapari, optimum distance between each antibiotic disc. The Barbari, Jakharana, Sirohi kids during the period Petri plates embedded with antibiotic discs are incubated at 37ºC for 24 hours. After incubation the Isolation of E. coli
results are read by measuring the zone of inhibition The fecal swabs collected were directly inoculated using a calibrated scale. The results are interpreted for each antibiotic by measuring the zone of simultaneously in duplicates for every sample inhibition and comparing the standard chart for the inoculums. Then the plates are incubated at 37ºC for 24 to 48 hours. After growth was observed the Results and Discussion
similar colonies were stained by the “grams” for Isolates were further assessed for the antibacterial examining the size, morphology and staining drug susceptibility by using disc diffusion method.
characteristics. The colony types, which were Out of 8 antibiotics, the isolates were found to be showing gram negative cocco bacilli, were further Oxytetracycline, Gentamicin, Ceftriazone, Cloxocillin On MacConkey’s agar only lactose fermenting (LF with inhibition zone diameter ranging between 10- +) colonies, which were pink in colour were isolated 15 mm. Chloramphenicol were highly sensitive for Evaluation of Moringa oleifera Seed Extracts for Antibacterial E. coli strains to producing the inhibition zone of 23- Ceftriazone, and Cloxocillin resulted found to be 26 mm and Ampicillin, Oxytetracycline, Gentamicin, intermediate the inhibition zone of 13-16 mm each.
Table 1: Antibiotic Sensitivity profiles of the field isolates of E. coli.
Chloram-
S.No Ampicillin Enrofloxacin OTC Gen tamicin Ceftriazone Cloxacillin phenicol
R =Resistant, I= Intermediate, S = Sensitive Antibiotic sensitivity profile of the test isolates of E. coli . Table 2: Effect of Antibiotics on different isolates of E. coli
Antibiotic
Resistant
Intermediate
Sensitive
R =Resistant, I= Intermediate, S = Sensitive Table 3: Percentage of Resistance of the total isolates shown to different antibiotics
Antibiotic
Percentage
Antibiotic sensitivity test were done to study the inhibiting the in vitro growth of E. coli. trend in susceptibility to commonly used antibiotics Chloramphenicol has given the lowest percentage in the field. Only those screened using bfpA gene of resistance at zero and the next best was were subjected to antibiotic sensitivity test. Some Enrofloxacin at 34 percent. So, it’s a serious issue of the commonly used antibiotics like Ampicillin, that out of the commonly used 7 antibiotics, only and Cloxacillin were giving high percentage of two was effective, which gives clue about the resistance at the rate of 75 percent and 100% emerging multi-drug resistant E. coli strains.
respectively. This is due to the unscrupulous use Gupta et al (2000) subjected drug sensitive test for of such antibiotics leading to high resistance. About isolates of faecal sample of effective kids, with the 67 percent of the isolates were resistant to range of modern antibiotics and samples were Oxytetracycline, Gentamicin, and Ceftriaxone. Only sensitive to Amikacin, Cefotaxime, Ceftazidime, antibiotic groups like fluoroquinolones and the Ceftrioxone, Cefuroxime, Cefadroxyl, Kanamycin Chloramphenicol have fared well in effectively Evaluation of Moringa oleifera Seed Extracts for Antibacterial Jose et al, (2001) isolated quinolone resistance
pathogenic and non-pathogenic Escherichia coli rearing. Intas Polivet, 1: 178-179.
strains from healthy ruminants. In cattle, 5.9% of the Hodgson J.C. (1994). Diseases due to
strains were resistant to nalidixic acid and 4.9% Escherichia coli in shepp. In: C.L. Gyles (Ed.).
were resistant to enrofloxacin and ciprofloxacin, Escherichiu coli in domestic animals and whereas in sheep and goats only 0.5% and 1.4%, humans. CAB International. UK, pp. 135- 150.
respectively, of the strains were resistant to nalidixic acid and none to fluoroquinolones. Most of the Joe Rook (2007), Diarrhea (scours) in small
strains resistant to quinolones were non-pathogenic ruminant, Last updated by Susan Schoenian strains isolated from cattle. But in the current study the resistance to fluoroquinolones was as high as Jose A. Orden, Jos A., Ruiz-Santa-
34 percent, making it a concern, for the future use Quiteria, Dolores Cid, Rosa Díez, Susana
Martínez and Ricardo de la Fuente (2001)
Kumar et al, (2003) isolated nineteen different Quinolone resistance in potentially pathogenic serotype of E. coli from faecal sample of diarrheic and non-pathogenic Escherichia coli strains cow and calves. Multiple drug resistance was a isolated from healthy ruminants. Journal of common feature belonging to isolates from all Antimicrobial Chemotherapy, 48: 421-424.
categories of animals. Highest resistance was Kumar A., Vaid R.K., Rana R., Vihan VS
against ampicillin and oxytetracycline.
and Gupta VK (2001) Descriptive epidemiology
Conclusion
of colibacillosis outbreak in Barbari kids under semi-intensive system, Indian J. of Animal Biochemical studies like IMViC, TSI, Oxidase, Sciences, 72; 233-236.
Catalase and Nitrate tests identified different strains of E. coli from clinical cases of diarrhea. Only Kumar Monoj and Richa (2003) Prevalence
Chloramphenicol showed good percentage of and Antibiogram of E. coli serotypes in healthy affectiveness against the pathogenic E. coli and diarrhoeic Cattle in Jabalpur (MP), Indian isolates, Ampicillin, Enrofloxacin, Oxytetracycline, Vet. Med. Jour., 27: 235-237.
Gentamycin, Ceftrioxone, and Cloxacillin antibiotics Shrestha J.N.B. and Galal E.S.E. (2011).
showed alarmingly low level of effectiveness in Conservation of goat genetic resources. Small inhibiting the growth of pathogenic E. coli in vitro.
Ruminant Research, (in press).
References
Vihan V.S., singh S.V. and Sing N. (1990)
Prevalence, pathogenecity and serotypes of E. Gupta V.K., Kumar A. and Singh N. (2000)
coli associated with diarrhoea in newborn kids, Indian J. Animals Science, 60, 793-795.

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