In vitro Antimicrobial Activity of Nanoencapsulated Bromelain against Bacteria Isolated from Milk of Dairy Goats with Sub-clinical Mastitis in Thika East Sub-county, Kenya

Main Article Content

Mahlangu Precious
Kagira John
Maina Naomi


goats is managed by a variety of antibiotics. Due to the emergence of antibiotic resistance, there is need for development of new antimicrobial agents. In the current study, the in vitro activity of nanoencapsulated bromelain, using bromelain extracted from the pineapple fruit, Annanus comosus was investigated against bacteria isolated from milk of dairy goats with sub-clinical mastitis. Nanoencapsulation of bromelain was done using the ionic gelation method of chitosan nanoparticles with sodium trypolyphosphate as the cross linking agent.  In this study, the agar well diffusion method was used to test for antimicrobial activity while the broth microdilution method was used to test for the Minimum Inhibitory Concentration (MIC). The isolates used were Staphylococcus aureus, Coagulase Negative Staphylococci, Serratia spp., Klebsiella spp., Enterobacter spp., Citrobacter spp. and Escherichia coli isolated from milk of dairy goats with sub-clinical mastitis in Thika East Sub-county, Kenya. The agar well diffusion method showed that bromelain and nanoencapsulated bromelain had antimicrobial activity. All of the tested bacteria were sensitive to extracted bromelain at 5 mg/ml and less. The tested bacteria were less sensitive to commercial bromelain (57.1%) at 5 mg/ml and less. The MIC of nanoencapsulated bromelain against Enterobacter spp., Citrobacter spp., Serratia spp. and Coagulase Negative Staphylococci was 25 µg/ml, while that of Escherichia coli was 50 µg/ml. The MIC of nanoencapsulated bromelain against Klebsiella spp. and Staphylococcus aureus was 200 µg/ml. The low MICs recorded in this study shows that nanoencapsulated bromelain has high antimicrobial potential which warrants further in vivo studies in dairy goats to determine its efficacy against sub-clinical mastitis.

In vitro, efficacy, bromelain, mastitis, Staphylococci, E. coli.

Article Details

How to Cite
Precious, M., John, K., & Naomi, M. (2020). In vitro Antimicrobial Activity of Nanoencapsulated Bromelain against Bacteria Isolated from Milk of Dairy Goats with Sub-clinical Mastitis in Thika East Sub-county, Kenya. Asian Journal of Research in Animal and Veterinary Sciences, 5(3), 33-40. Retrieved from
Original Research Article


Contrares A, Rodriguez JM. Mastitis: Comparative Etiology and Epidemiology Journal of Mammary gland Biol Neoplasia. 2011;16:339-356.

Arguello A. Trends in goat research, a review, Journal of Applied Animal Research. 2011;39(4):429-434.

Stuhr T, Aulrich K. Intramammary infections in dairy goats: Recent knowledge and indicators for detection of subclinical mastitis, Agriculture and Forestry Research. 2010;4(60):267-280.

Ndegwa EN, Mulei CM, Munyua SJ. The prevalence of subclinical mastitisin dairy goats in Kenya. Journal of the South African Veterinary Association. 2000;71(1): 25-7.

Mbindyo MC. To characterise dairy goats production in Mount kenya Region; determinition of prevalence and risk factors of subclinical mastitis and antibiotic sensitivity of the isolates, University of Nairobi MSc Thesis; 2014.

Makau L. Prevalence of mastitis and associated risk risk factors in dairy goats in Machokos County, Kenya, University of Nairobi MSc Thesis; 2017.

Mahlangu P, Maina N, Kagira J. Prevalence, risk factors, and antibiogram of bacteria isolated from milk of goats with subclinical mastitis in Thika East Subcounty, Kenya, Journal of Veterinary Medicine. 2018;8.

[Article ID 3801479]

Mbilu TJNK. Status of mastitis in lactating goats at Sokoine University of Agriculture and neighbouring small holder farms in Morogoro Municipality, Tanzania, Livestock Rural Development. 2007;19(3): 54-60.

Pirzada M, Marhi KK, Kamboh AA, Rind R, Abro H, Lakho SA, Bhutto KR, Huda N . Prevalence of subclinical mastitis in dairy goats caused by bacterial species, Journal of Animal Health and Production. 2016;4: 55-59. DOI:10.14737/journal.jahp/2016/

Hristov K, Parvanov P, Kashamov B, Pepovich R, Nikolov B. Risk factors influencing the prevalence of subclinical mastitis in goats. Scientific Works, Series C, Veterinary medicine. 2016;LX(1):53-57.

Ajibade VA, Akinruli FT, Ilesanmi TM. Antibacterial screening of crude extract of oven-dried pawpaw and pineapple. International Journal of Scientific and Research Publications. 2015;5(11).

Reis CP, Neufeld RJ, Ribeiro AJ, Veiga F. Nanoencapsulation I. Methods for preparation of drugloaded polymeric nanoparticles, Nanomedicine: Nanotechnology, Biology and Medicine. 2006;2:8– 21.

Bromelain Monograph. Alternative medicine review. 2010;15(4):361-368.

Otani S, Ishikawa Y, Kanno K, Ishizawa N. Effect of bromelain on bovine mastitis. Journal of Veterinary Medicine (Japan). 1989;812:172–175.

Contreras A, Paape MJ, Miller RH, Corrales JC, Luengo C, Sãnchez A. Effect of Bromelain on milk yield, milk composition and mammary health of dairy goats. Tropical Animal Health and Production. 2009;41:493-498.

Eshamah H, Han I, Naas H, Rieck J, Dawson P. Bactericidal Effects of Natural Tenderizing Enzymes on Escherichia Coli and Listeria monocytogenes. Journal of Food Research. 2013;2(1):8.

Ali AA. Antimicrobial Effects of Crude Bromelain Extracted from Pineapple Fruit (Ananas comosus (Linn.) Merr.).Advances in Biochemistry. 2015;3(1):1.

Ashik AA, Vishnu PV, Gayathri R, Geetha RV. Evaluation of anti microbial activity of pineapple extract against selected microbes. International Journal of Pharmaceutical Sciences Review and Research. 2016;39(1):277–278.

Ra JC, Kang KS, Park YH, Han HJ, Lee JE. Composition for preventing or treating mastitis of dairy cattle - , World Intellectual Property Organisation, International Publication W02003057233A1, Google Patents; 2002.

Tachaboonyakiat W. Antimicrobial applications of chitosan, Chitosan Based Biomaterials. 2017;2.

Ma Z, Garrido-Maestu A, Jeong KC. Application, mode of action, and in vivo activity of chitosan and its micro- and nanoparticles as antimicrobial agents: A review. Carbohydrate Polymers. 2017;176: 257–265.

Kahiro SK, Kagira JM, Maina N, Karanja SM, Njonge FN. Enzymatic Activity of Bromelain from Crude Extracts of Crown, Peels and Stem of Pineapples from Different Agro-ecological Zones of Thika Region, Kenya, Asian Journal of Biotechnology and Bioresource Technology. 2017;1(2):1-6.
[Article no.AJB2T.34314]

Fan W, Yan W, Xub Z, Nia H. Formation mechanism of monodisperse, low molecular weight chitosan nanoparticles by ionic gelation technique, Colloids and Surfaces B: Biointerfaces. 2012;90: 21–27.

Gan Q, Wang T, Cochrane C, McCarron P. Modulation of surface charge, particle size and morphological properties of chitosan–TPP nanoparticles intended for gene delivery. Colloids and Surfaces B: Biointerfaces. 2005;44(2);65-73.

Saha P, Goyal AK, Rath G. Formulation and evaluation of chitosan-based ampicillin trihydrate nanoparticles. Tropical Journal of Pharmaceutical Research. 2010;9(5).

Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review, Journal of Pharmaceutical Analysis. 2015;6(2016): 71–79.

CLSI, Performance Standards for Antimicrobial Susceptibility Testing, Twenty fifth information Supplement, Clinical and Laboratory Standards Institute, 950 West Valley Road, Suite 2500,Wayne, Pennsylvania 19087, USA; 2015.

Illanes A. Enzyme Production. In: Enzyme Biocatalysis. Brazil: Springer Science and Business Media V.B. 2008;57-106.

Zharfan RS, Purwono PD, Mustika A. Antimicrobial activity of pineapple (Ananas comosus L. Merr) Extract against multi-drug resistant Pseudomonas aeruginosa: An in vitro study, Indonesian Journal of Tropical and Infectious Disease. 2017;6(5): 118–123.

Bansode DS, Chavan MD. Evaluation of antimicrobial activity and phytochemical analysis of papaya and pineapple fruit juice against selected enteric pathogens International Journal of Pharm and Biosciences. 2013;4:1176–184.

Ali R, Al-Achkar K, Al-Mariri A, Safi M . Role of Polymerase Chain Reaction (PCR) in the detection of antibiotic-resistant Staphylococcus aureus, The Egyptian Journal of Medical Human Genetics. 2014; 15:293–298

Shweta P. Bromelain A Cysteine Protease: Helps To Reduce Infection Caused By Acinetobacter spp., A Nosocomial Pathogen. International Journal of Advanced Biotechnology and Research. 2014;5:976–2612.

Shiew PS, Fang YL, Adibah F, Majid A. In vitro study ofbromelain activity in artificial stomach juice and blood, in Proceedings of the 3rd International Conference on Biotechnology for theWellness Industry, PWTC; 2010.

Bernela M, Ahuja M, Thakur R. Enhancement of anti-inflammatory activity of bromelain by its encapsulation in katira gum nanoparticles. Carbohydrate Polymers. 2016;143:18–24.

Cheung RCF, Ng TB, Wong JH, Chan WY. Chitosan: An Update on Potential Biomedical and Pharmaceutical Applications, Marine Drugs. 2015;13: 5156-5186.