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Bacterial and Antibiotic Profile of Mastitic Milk of Beetal Goat in District Jhelum

PJZ_53_6_2151-2155

Bacterial and Antibiotic Profile of Mastitic Milk of Beetal Goat in District Jhelum

Waqar Ali1,3*, Muhammad Yaqoob2, Abida Arshad1, Guifu Dai3, Safir Ullah Khan1, Muhammad Arshad Malik4, Imran Ullah1 and Ihsan Ullah1

1Department of Zoology, PMAS-Arid Agriculture University, Rawalpindi, Pakistan

2Department of Clinical Studies PMAS-Arid Agriculture University, Rawalpindi, Pakistan

3School of Life Sciences, Zhengzhou University, Henan, China.

4Department of Biological Sciences, International Islamic University, Islamabad, Pakistan

ABSTRACT

A total of 514 milk samples collected independently from beetal goats were tested for prevalence of clinical and subclinical mastitis through surf field mastitis test. The bacterial isolates identified through different biochemical test and colony morphology from positive milk sample were Staphylococcus aureus (25.4%), Escherichia coli (20.3%), Streptococcus (16.9%) in which (6.7%) Streptococcus agalactiae, and (10.1%) other streptococcus species, Pseudomonas aeruginosa (15.2%), Salmonella (13.5%) and Klebsiella pneumonia (8.4%). Staphylococcus aureus was found most prevalent pathogen that causes mastitis in goats. The overall prevalence of goat mastitis was recorded (11.67%). Risk factor like age, lactation stage, parity number and unhygienic condition showed statistically significant association with the prevalence of mastitis in beetal goats. Antibiotics sensitivity profiles of investigated bacteria showed that these were most sensitive to gentamycin, ciprofloxacin and amoxicillin.


Article Information

Received 12 October 2020

Revised 29 October 2020

Accepted 19 November 2020

Available online 13 September 2021

Authors’ Contribution

MY and AA designed the study. WA performed experiments and wrote the manuscript. Imran U and Ihsan U helped in sampling and experiments. MAM did analysis. GD provided assistance and research guideline. SUK helped in manuscript finalizing.

Key words

Mastitis, Bacterial isolates, Surf field mastitis test, Biochemical test, Risk factor, Antibiotic sensitivity

DOI: https://dx.doi.org/10.17582/journal.pjz/20201012061011

* Corresponding author: waqar.uaar757@gmail.com

0030-9923/2021/0006-2151 $ 9.00/0

Copyright 2021 Zoological Society of Pakistan



INTRODUCTION

Goats are mainly reared for the purpose of milk and meat all over the world and considered one of the ancient housekeeping species (Hale et al., 2011). It has been evaluated that 460 million goats are present in the world, which produce almost 4.50 million tons of milk and 1.20 million tons of meat approximately (Hirpa and Abebe, 2008). In Pakistan, entire population of goats recorded is 63.1 million that produce 0.779 million tons milk and 0.629 million tons meats (Anonymous, 2003). Beetal breed is one of the described breed out of 36 goat breeds bear great significance in milk production via FAO in 2006. Beetal goat has 4.2 million populations in Pakistan, constitute 7.8% of the whole goat population of Pakistan (GOP, 2008). Mastitis is the inflammatory state of the udder, described by chemical and physical changes occur in milk quality (Nazifei et al., 2011). Mastitis has been considered the most imperative and costly disease of milk industry, it causes severe damage to dairy industry in the form of decreased milk production, cost of treatment, increasing labor, and early elimination (Miller et al., 1993). The most essential modifications occur in milk of animals during mastitis include discoloration, appearance of small clots and existence of large amount of leukocytes. There are three main sub-types of mastitis that is clinical mastitis, sub-clinical mastitis and chronic mastitis. (Anonymous, 2003). In clinical mastitis milk color changes and clots also appears in the milk. Subclinical mastitis can be diagnosed by different laboratory test including microbial investigation, California mastitis test, surf field mastitis test, somatic cell count and white side test (Wakoya et al., 2006). In chronic mastitis, blisters and granulomas appear in the tissue of mammary gland that can reduce milk production while such cases are not often diagnosed and only visible at abattoirs (Hulten et al., 2004).

Although Staphylococcus species are considered to be the most frequent causative agent for mastitis, Streptococcus species, Pseudomonas aeruginosa, corynebacteria, and fungi have also been reported to cause mastitis in ruminants (Contreras et al., 2007). Mastitis disease cannot be completely removed from animals herd, but their occurrence could be reduced to minute level. Antimicrobial therapy considers to be an important component of mastitis control in dairy production systems (Oliver et al., 2011). Antibiotics such as lactam, macrolide, flouroquinolones, penicillin, nafcillin, and dihydro-streptomycin have been found most effective in decreasing the liability of mastitis bacteria (Chaffer et al., 2003). In Punjab province a total loss of Rs. 240 million per annum have been recorded due to this disease. The aim of this study is to identify the bacterial isolates and to determine their drug sensitivity.

MATERIALS AND METHODS

In this study, 514 milk samples were collected aseptically from beetal goats in four tehsils Jhelum, Sohawa, Dina and Pind dadan khan of Jhelum district. The teats were cleaned with 70% ethyl alcohol cotton swab. First three milk streaks were discarded then 5 to 10ml of milk was collected aseptically in a sterile beaker. The clinical mastitis was recognized by palpation of udder and visual observation of teat lesions. Milk samples were collected according to the national mastitis council (NMC, 1990). Parameters like changes in milk color, presence of grossly visible substances, swelling, pain, redness, udder hardening, and obstruction of the teats were assessed for detection of clinical mastitis. Sub-clinical mastitis was diagnosed by surf field mastitis test (Muhammad et al., 1995). From goats teat milk was stripped into well of the paddle plate and an equal amount of surf solution was added. The paddle plate was then rotated for 10 seconds to proper mix the surf solution and milk. After 25 seconds, variations in the milk were detected. Development of clots in milk and gel like flakes were suggestive of the disease. The positive milk samples were cultured on nutrient agar media, blood agar enriched media and selective macConkey agar media for bacterial identification according to standard procedures (Quinn, 1994). For the identification of gram negative bacteria, the triple sugar iron, Indole, Urease, Citrate and Oxidase test were performed, while for gram positive cocci chain and cocci cluster catalase, coagulase, bile test and camp test were conducted (Balows et al., 1991). The isolates were tested for antibiotic sensitivity against various antibiotics including gentamycin, ciprofloxacin, oxytetracycline, amoxicillin, sulfa-trimethoprim and kanamycin according to disc diffusion method described by (Bauer et al., 1966). The inhibition zone around every antibiotic disc was measured according to national committee for clinical laboratory standards (Watts et al., 2018). Several risk factors potentially associated with prevalence of mastitis were recorded. These factors included lactation period, parity, age and rearing area sanitation. First, the data of these factors were arranged for univariate logistic regression analysis to determine the individual risk factor effect on mastitis. The factors that ensured p value <0.05 during the univariate analysis were preferred as possible risk factor for multivariate logistic model analysis. The final logistic model was constructed with a significant level of p≤0.05.

RESULTS AND DISCUSSION

Table I shows prevalence of mastitis in different tehsils and union councils of district Jhelum. Overall prevalence of mastitis was 11.67%. Union council Sauwal of tehsil Pind Dadan Khan showed 12.32% prevalence. Union council Kala Gujjran of tehsil Jhelum showed 8.88%, union council Khukha of tehsil Dina showed 15.23% prevalence and union council Lehri of tehsil Sohawa showed 10.93% prevalence of mastitis. Our results agreed well with previously reported (Zamin et al., 2010) prevalence of 14.5%.

 

Table I. Mastitis Prevalence in different tehsils and union councils of district Jhelum.

Union councils (Tehsil)

No of positive/No of samples tested

Percentage of positive

Sauwal (Pind dadan khan)

18/146

12.32%

kala gujjran (Jhelum)

12/135

8.88%

Khukha (Dina)

16/105

15.23%

Lehri (Sohawa)

14/128

10.93%

Total

60/514

11.67%

 

Risk factors

Table II shows the risk factors associated with mastitis in beetal goat. It was observed goats of 2 and 3 years of age had no significant association with disease development. On the other hand, goats in the age of 4, 5 and 6 years had mastitis 22%, 7.3% and 1.9% in comparison with control 24 (9.2%) showing (P<0.05) significant association. It increases the odds of mastitis 18.391 (3.862-87.576), 111.189 (17.047-725.221) and 21.645 (1.936-241.956) times, with 95% confide-nce interval. In combination with other risk factors its effect become more prominent as the odd ratio increased 19.465 (4.264-88.850) times. Mekibib et al. (2010) showed that the univariate analysis of possible risk factors was extremely higher (P<0.05) with increasing age and the odds of mastitis increased after 4 year of age. This result is similar to (Amin et al., 2011) that the odd ratio of mastitis increased to 52 (33.5, 69.97) in 5 years of age.

Beside that it was observed that first and second month of lactation stage were significantly associated to mastitis. Odd ratio of lactation stage during initial months increases the odds of mastitis 37.074 (5.758-238.697) times with 95% confidence interval. Mastitis recorded in three and four months of lactation stage were 12 (4.6%) and 5 (1.9%) showing increased in the odds of mastitis 77.033 (12.784-464.181 and 10.019 (1.937-51.812) times. After multiple logistic regression analysis, it was found to have significant association with mastitis, as increased the adjusted odd ratio (AOR) 99.311 (16.902-583.511) and 12.814 (1.971-48.852) times. On the base of this analysis, it is concluded that early stage of lactation is a potential risk factor for mastitis and the prevalence of mastitis was also high while prevalence decreased with increasing lactation period. The finding of current study coincides to the outcomes of (Svensson et al., 2006) that 31% of mastitis was recorded in first seven days of lactation, while maximum prevalence reported in period of 7 to 30 days after parturition.

It is evident from statistical analysis that the odds of mastitis increases 16.635 (2.299-120.371) and 25.277 (2.946-216.866) times with 95% confidence interval. The finding of this study support the earlier research work with solid significant association of mastitis with increasing parity (Mekibib et al., 2010). Finding of increasing prevalence of CM and SCM with the increasing number of parity are also in agreement with mostly previous reported research work (Amin et al., 2011). The prevalence of mastitis has increased gradually with increasing parity number of the studied goats. This is in close agreement to other findings (McDougall et al., 2002).

 

Table II. Univariate and multivariate logistic regression analysis of possible risk factors associated with mastitis in beetal goats.

Factors

Control (%)

Mastitis (%)

Univariate analysis

Multivariate analysis

OR (95%)

P-value

OR (95%)

P-value

Age

2 years

3 years

4 years

5 years

6 years

75 (28.8%)

82 (31.5%)

24 (9.2%)

11 (4.2%)

8 (3.1%)

5 (1.9%)

9 (3.5%)

22 (8.5%)

19 (7.3%)

5 (1.9%)

.00

1.570 (.319-7.732)

18.391 (3.862-87.576)

111.189 (17.047- 725.221)

21.645 (1.936-241.956)

.579

.000

.000

.013

.00

1.969 (.426-9.113)

19.465 (4.264-88.850)

91.745 (14.491-580.867)

21.140 (1.813-246.450)

.386

.000

.000

.015

Lactation stage

1 month

2 months

3 months

4 months

5 months

10 (3.8%)

14 (5.4%)

30 (11.5%)

71 (27.3%)

75 (28.8%)

14 (5.4%)

26 (10.0%)

12 (4.6%)

5 (1.9%)

3 (1.2%)

.00

37.074 (5.758-238.697)

77.033 (12.784-464.181

10.019 (1.937-51.812)

1.346 (.238-7.624)

.000

.000

.006

.737

.00

23.823 (4.257-133.313)

99.311 (16.902-58.511)

12.814 (1.971-48.852)

1.159 (.207-6.472)

.000

.000

.005

.867

Parity

1

2

3

4

5

43 (16.5%)

58 (22.3%)

64 (24.6%)

24 (9.2%)

11 (4.2%)

3 (1.2%)

8 (3.1%)

12 (4.6%)

23 (8.8%)

14 (5.4%)

.00

3.007 (.409-22.086)

2.228 (.323-15.383)

16.635 (2.299-120.371)

25.277 (2.946-216.866)

.279

.416

.005

.003

.00

2.307 (.348-15.280)

1.666 (.266-10.417)

12.868 (2.016-82.116)

20.295 (2.665-154.542)

.386

.585

.007

.004

Rearing area

Hygienic

Unhygienic

108 (41.5%)

92 (35.4%)

24 (9.2%)

36 (13.8%)

.00

2.989 (0.965-9.262)

.058

 

Table III. Antibiotic sensitivity pattern of isolates against antibiotics.

Antibiotics sensitivity

Bacterial isolates

S. aureus

E. coli

Streptococcus

P. aeruginosa

Salmonella

K. pneumonia

Gentamycin

92%

87%

86%

80%

78%

77%

Ciprofloxacin

88%

74%

82%

76%

74%

72%

Amoxicillin

82%

74%

75%

72%

70%

68%

Oxytetracycline

80%

70%

68%

70%

66%

64%

Sulpha-trimethoprim

74%

70%

68%

65%

60%

54%

Kanamycin

70%

66%

60%

58%

54%

50%

 

It was further found that mastitis in goats that were living in unhygienic area were 36 (13.8%) among healthy control of same group showing (P<0.05) significant association. It increases the odds of mastitis 2.989 (0.965-9.262) times. However same result was described by (Bergonier et al., 2003) that environmental factors like humidity, litter, wet bedding, forage, mixed reservoirs, housing, clusters, and feed stuffs are the major determinant of mammary pathology in dairy goats. Similar results also concluded by (Amin et al., 2011) and found the prevalence higher in a goats rearing in unhygienic area and farming goats than those are managed under subsistence system.

Antibiotic sensitivity

Highest prevalence of Staphylococcus aureus was 25.4% followed by Escherichia coli 20.3%, Streptococcus 16.9%, and Streptococcus agalactiae 6.8% other streptococcus 10.1%, Pseudomonas aeruginosa 15.2%, Salmonella 13.5% and Klebsiella pneumonia 8.4% in lowest prevalence. The similar prevalence has also been recorded by (Contreras et al., 2007). However, relative difference in prevalence and bacterial isolates of goat mastitis may be due to difference in geographic regions. The main bacterial species involved in bacteriology of both clinical and sub clinical mastitis of dairy goat are Staphylococcus aureus, Escherichia coli, Streptococcus, Salmonella, Pseudomonas species and Klebsiella pneumonia. Similarly, the study conducted by Wakoya et al. (2006) concluded that staphylococcus species was the major pathogen identified in Ethiopia, while Escherichia coli, Streptococcus species, Pseudomonas, Klebsiella pneumonia, corynebacterium, bacillus and salmonella species were also identified.

Table III shows the antimicrobial sensitivity of the identified bacteria isolated from goat’s milk samples against antibiotics like gentamycin, ciprofloxacin, oxytetracycline, amoxicillin, sulfa-trimethoprim and kanamycin. These finding are similar to those of (Anjum et al., 2010) who reported that gentamicin, ciprofloxacin, amoxicillin norfloxacin, and kanamycin were most effective among the 13 antibiotics tested in vitro. However, (Islam et al., 2012) reported that gentamicin treatment of goats and sheep with sub-clinical mastitis increased milk production significantly. The response of host immune system was a significant factor to the causative bacteria for the austerity and regulation of mastitis (Bannerman et al., 2009). The probable justification for resistance of bacterial organisms to the existing antibiotics could be due unselective and prolonged use of antibiotics by individual farmers without prescription of professionals.

CONCLUSION

This study will be useful for veterinarians and researcher for the medication of mastitis by using drug of choice according to the occurrence of pathogens in these areas.

Statement of conflict of interest

The authors have declared no conflict of interest.

REFERENCES

Amin, M.A., Samad, M.A. and Rahman, A.A., 2011. Bacterial pathogens and risk factors associated with mastitis in black Bengal goats in Bangladesh. Bangladesh J. Vet. Med., 9: 155-159. https://doi.org/10.3329/bjvm.v9i2.13458

Anjum, A.A., Qamar, M.F. and Ali, M.A., 2010. Bacteriology of mastitic milk of cattle and in vitro antibiogram of the pathogens isolated, in district Hafizabad, Pakistan. Biologia, 56: 117-122.

Anonymous, 2003. Mastitis in dairy cows. Department of Animal Science, Faculty of Agricultural and Environmental Sciences, McGill University, Anne de Bellevue, Canada. pp. 12.

Balows, A., Housher, W.J., Herrman, K., Isenberg, H. and Shadoms, H., 1991. Manual of clinical microbiology, 5th Ed. American Society of Microbiology, Washington DC, USA.

Bannerman, D.D., Rinaldi, M., Vinyard, B.T., Laihia, J. and Leino, L., 2009. Effects of intramammary infusion of cis-urocanic acid on mastitis-associated inflammation and tissue injury in dairy cows. Am. J. Vet. Res., 70: 373-382. https://doi.org/10.2460/ajvr.70.3.373

Bauer, A.W., 1966. Antibiotic susceptibility testing by a standardized single disc method. Am. J. clin. Pathol., 45: 149-158. https://doi.org/10.1093/ajcp/45.4_ts.493

Bergonier, D., De Cremoux, R., Rupp, R., Lagriffoul, G. and Berthelot, X., 2003. Mastitis of dairy small ruminants. Vet. Res., 34: 689-716. https://doi.org/10.1051/vetres:2003030

Chaffer, M., Leitner, G., Zamir, S., Winkler, M., Glickman, A., Ziv, N. and Saran, A., 2003. Efficacy of dry-off treatment in sheep. Small Rumin. Res., 47: 11-16. https://doi.org/10.1016/S0921-4488(02)00194-3

Contreras, A., Sierra, D., Sanchez, A., Corrales, J.C., Morco, J.C., Paape, M.J. and Gonzalo, C., 2007. Mastitis in small ruminants. Small Rumin. Res., 68: 145–153. https://doi.org/10.1016/j.smallrumres.2006.09.011

Hale, M., Coffey, L., Spencer, T. and Pressman, A., 2011. Small-Scale livestock production. ATTRA.

Hirpa, A. and Abebe, G., 2008. Economic significance of sheep and goats.

Hulten, F., Persson, A., Eliasson-Selling, L., Heldmer, E., Lindberg, M., Sjogren, U. and Ehlorsson, C.J., 2004. Evaluation of environmental and management related risk factors associated with chronic mastitis in sows. Am. J. Vet. Res., 65: 1398-1403. https://doi.org/10.2460/ajvr.2004.65.1398

Islam, M.R., Ahamed, M.S., Alam, M.S., Rahman, M.M., Sultana, T., Roh, Y.S. and Kim, B., 2012. Identification and antibiotic sensitivity of the causative organisms of subclinical mastitis in sheep and goats. Pak. Vet. J., 32: 179-182.

McDougall, S., Pankey, W., Delaney, C., Barlow, J., Murdough, P.A. and Scruton, D., 2002. Prevalence and incidence of subclinical mastitis in goats and dairy ewes in Vermont, USA. Small Rumin. Res., 46: 115–121. https://doi.org/10.1016/S0921-4488(02)00191-8

Mekibib, B., Furgasa, M., Abunna, F., Megersa, B. and Regassa, A., 2010. Bovine mastitis, Prevalence, risk factors and major pathogens in dairy farms of Holeta Town, Central Ethiopia. Vet. World, 3: 397-403. https://doi.org/10.5455/vetworld.2010.397-403

Miller, G.Y., Bartlett, P.C., Lance, S.E., Anderson, J. and Heider, L.E., 1993. Costs of clinical mastitis and mastitis prevention in dairy herds. J. Am. Vet. med. Assoc., 202: 1230–1236.

Muhammad, G., Athar, M., Shakoor, A., Khan, M.Z., Rahman, F. and Ahmad, M.T., 1995. An inexpensive new tool for evaluation of wholesomeness of fresh milk. Pak. J. Fd. Sci., 5: 91-93.

National Mastitis Council (US) Research Committee. 1990. Microbiological procedures for the diagnosis of bovine udder infection. National Mastitis Council.

Nazifei, S., Haghkhah, M., Asadi, Z., Ansari-Lari, M., Tabandeh, M.R., Esmailnezhad, Z. and Aghamiri, M., 2011. Evaluation of sialic acid and acute phase proteins in clinical and subclinical bovine mastitis. Pak. Vet. J., 31: 55-59.

Oliver, S.P., Murinda, S.E. and Jayarao, B.M., 2011. Impact of antibiotic use in adult dairy cows on antimicrobial resistance of veterinary and human pathogens, a comprehensive review. Foodborne Pathog. Dis., 8: 337-355. https://doi.org/10.1089/fpd.2010.0730

Quinn, P.J., 1994. Clinical veterinary microbiology (No. SF 780.2. C54).

Svensson, C., Nyman, A.K., Waller, P.K. and Emanuelson, U., 2006. Effects of housing, management, and health of dairy heifers on first lactation udder health in southwest Sweden. J. Dairy Sci., 89: 1990–1999. https://doi.org/10.3168/jds.S0022-0302(06)72266-4

Wakoya, A., Molla, B., Belihu, K., Kleer, J. and Hildebrandt, G., 2006. A cross-sectional study on prevalence, antimicrobial susceptibility pattern and associated bacterial pathogens of goat mastitis. Int. J. appl. Res. Vet. Med., 4: 169–176.

Watts, J.L., Sweeney, M.T. and Lubbers, B.V., 2018. Antimicrobial susceptibility testing of bacteria of veterinary origin. Antimicrob. Resist. Bacteria Livest. Companion Anim., 1: 17-32. https://doi.org/10.1128/9781555819804.ch2

Zamin, Ali., Muhammad, G., Ahmad, T., Khan, R., Naz, S., Anwar, H. and Usama, A.R., 2010. Prevalence of caprine sub-clinical mastitis, its etiological agents and their sensitivity to antibiotics in indigenous breeds of Kohat, Pakistan. Pak. J. Life Soc. Sci., 8: 63-67.

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