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2007, Vol 2 No 1, Article 11

 

Bacterial Isolation and Drug Sensitivity Pattern of

 

Bovine Mastitis in Kashmir

 

M. H. Tarfarosh1, S. G. J. Qadri2, A. M. Qurashi3

Institute of Animal Health & Biological Products, Zakura

1Research Assistant, 2Assistant Parasitology, 3Joint Director

 


ABSTRACT

A retrospective analysis of the bovine milk samples screened for causative agents of mastitis at the Institute of Animal Health and Biological Products, Zakura from the year 1997 to 2003 was undertaken. Four hundred and thirty five bovine milk samples received over a period of six years from various places of the Kashmir valley were subjected to bacterial isolation and antibiotic sensitivity. Staphylococci were present in 72% to 76.3%, Streptococci in 12.5% to 14% and mixed infection in 9.7% to 11.2% of the samples tested. The periodic prevalence of Staphylococcal and Streptococcal mastitis and shift in the antibiograms of isolates over the period of these six years was recorded.

INTRODUCTION

Mastitis is the most important disease of high yielding dairy cows. Economic loss to the poor farmers due to loss of milk during current and future lactations is enormous. Wide range of microbes responsible for the disease contribute differently in various climatic zones and managemental practices. The sensitivity pattern of different bacteria to antibiotics also changes over a period of time.

MATERIALS AND METHODS

The samples collected / received from the field , were subjected to cultural examination using standard laboratory procedures of inoculating the samples in nutrient broth overnight and then transferring to solid media i.e. nutrient agar, blood agar, differential agar, strepto-staphylococcal agar (Hi-media). The isolated colonies were studied for the staining character, morphology and hemolytic activity. Selected isolates were then studied for bio-chemical characteristics and identified as Staphylococci or Streptococci as per the Bergey’s Manual for Determinative Bacteriology. The antibiotic disks used in the sensitivity studies are given in Table 1.

 


Table 1

Antibiotics used in the study and their concentrations

Antibiotic  disc  used

Abbreviation

Concentration (µg)

Ampicillin

A

10

Chloramphenicol

C

30

Ciprofloxacin

Cf

5

Cloxacillin

Cx

5

Clorotetracycline

Ct

30

Doxycycline  hydrochloride

Do

30

Erythromycin

E

15

Gentamicin

G

10

Kanamycin

K

30

Lincomycin

L

10

Norfloxacin

Nx

10

 

The Staphylococcus and Streptococcus isolates were grown in separate nutrient broths and aseptically streaked on sterile nutrient agar plates. The antibiotic discs were placed five minutes later selectively with the help of an antibiotic disc dispenser in a clockwise direction. The charged plates were incubated overnight at 37ş C by keeping the plates in an inverted position. (Kirby et al, 1966).
For determining the sensitivity of each antibiotic for therapeutic purpose, the diameter of zone of inhibition including the disc were measured in millimeters (mm). The organisms were considered sensitive to a particular drug when no growth could be seen in an area of 7mm diameter around the antibiotic discs.

RESULTS AND DISCUSSION

(a) Prevalence

The retrospective analysis was conducted from the years 1997-98 to 2002-03 (six years) wherein the cases of milk samples tested were 56, 100, 80, 71, 56 and 72 respectively.
The percentage, distribution of staphylococcal and streptococcal isolates recovered from mastitis milk over the period is depicted in Table 2.

Throughout the study period Staphylococcus was the major bacterial species associated with bovine mastitis followed by streptococci. These two organism thrive in abundance in the vicinity of the animals particularly under poor managemental practices.

The prevalence of Staphylococcal species in causing bovine mastitis was found to be 73%, 72.2%, 72.5%, 73.2%, 73% and 73% in the year 1997-98, 1998-99, 1999-2000, 2000-2001, 2001-2002 and 2002-2003 respectively, which is in agreement with earlier reports of Kalorey et al (1984), Singh et al, (1994) and Tarfarosh, (1996).

Similarly the prevalence of Streptococci was recorded in bovine mastitis as 12%, 13%, 13.7%, 14%, 12.5% and 13.8% in the year 1997-98, 1998-99, 1999-2000, 2000-2001, 2001-2002 and 2002-2003 respectively which is in conformity with the reports of Saini et al, (1994) and Tarfarosh, (1996).

The incidence of mixed infection was recorded as 10.7%, 10%, 11.2%, 11.2%, 10.7% and 9.7% in the year 1997-98, 1998-99. 1999-2000, 2000-2001, 2001-2002 and 2002-2003 respectively which falls in line with the observations of Konte et al, (1988), Kapur and Singh, (1980) and Tarfarosh, (1996).

(b) Drug sensitivity

The results of sensitivity test are presented in Tables 3 and 4.

The area of zone of inhibition with the diameter measured in mm and the quantum of sensitivity was interpreted as follows.

+++               >12mm                  Highly sensitive
++ 10-12mm Moderately sensitive
+ 7-10mm Relatively sensitive
0 < 7mm Resistant


During the year 1997-98, the drugs Cloxacillin and Chloramphenicol were found to be highly effective, but during subsequent years i.e. 1998-99, 1999-2000, 2000-2001, 2001-2002 and 2002-2003 Chloramphenicol, Gentamicin, Kanamycin, Ciprofloxacin were the drugs to which the cultures were highly and moderately sensitive but resistance was encountered for Cloxacillin, Ampicillin, Erythromycin.

 

Drugs of choice as per the Tables 3 and Table 4 appear to be Chloramphenicol, Gentamicin and Ciprofloxacin which fall in line with the works conducted by Babu et al (1980) Singh and Baxi (1983) Rehman and Boro (1991) Singh et al (1994) and Tarfarosh (1996 )

 


Table 2

Distribution of causative agents in the bovine milk samples

Year

Samples Received

Samples Positive for Mastitis

Causative agents

 

 

 

Staphylococci

%

Streptococci

%

Mixed

%

1997-98

56

54

41

73

7

12.5

6

10.7

1998-99

100

95

72

72

13

13

10

10

1999-2000

80

78

58

72.5

11

13.7

9

11.2

2000-01

71

70

52

73.2

10

14

8

11.2

2001-02

56

54

41

73

7

12.5

6

10.7

2002-03

72

70

53

76.3

10

13.8

7

9.7

 

 


Table 3

Year-wise Use of antibiotics and sensitivity patterns

Year

Isolates tested

Antibiotic discs used

Sensitive

Resistant

1997-98

54

C+++, CX+++, Cf++, G++

A, E, Nx

1998-99

95

C+++, K+++, G++, CF++, DO++

Cx, A, Nx

1999-2000

78

C+++, K+++, G++, CF++

A, Cx. Cl

2000-2001

70

C+++, G++, CF++, NX++

A, E, Cx, L

2001-2002

54

C+++, G++, Cf++

A, E, Nx

2002-2003

70

C+++, G++, Cf++

A, E, Nx

 

 

Table 4

Periodic changes in resistance pattern

Year

Drug

1997-98

Ac, Nx

1998-99

A, Cx

1999-2000

A, Cx, K

2000-2001

A, E, Cx

2001-2002

A, E, Cx

2002-2003

A, E, Nx

 

 

REFERENCES

  1. Babu et al (1980), Indian Vety. Journal 56(3) : 211-213
  2. Bar Moshe (1970), Refuat Vet 26(3) : 48-50 & 59-63 Dairy Science Abstract 32(3) 1970
  3. Kapur & Singh (1980), Indian Vet Journal 55(10) : 803-806
  4. Karoley et al (1984), Indian Journal of Animal Science 53(9) :961-963
  5. Kastle (1968), Dairy Science Abstract 30(1) : 258
  6. Kirby et al (1966), American Journal of Clinical pathology 45 : 493
  7. Konte et al (1988), Vet Bull (1988) 58 (7) : 4649
  8. Saini et al (1994), Indian J. of Dairy Science 47(2) : 103-106
  9. Sarma & Boro (1981), Journal of Animal Health 19(1) : 47-49
  10. Singh et al (1994), Symposium on Meat & Milk Industry Proceedings at CCS, HAU, Hissar
  11. Singh & Baxi (1983), Indian Vety. Journal 59(3) : 197-198
     

 


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