Antimicrobial Activity of Algerian Honey on Subclinical Mastitis Pathogens Isolated from Goat's Milk Susceptibility Resistant Susceptibility Resistant Susceptibility Resistant Susceptibility Resistant Susceptibility Resistant

Antimicrobial activity of Algerian honey on subclinical mastitis pathogens isolated from goat's milk, Veterinary World 7(4): 248-252. Introduction infected wounds [5-9]. Note that honey has no unique composition but various types could be collected Mastitis inflammation of the mammary gland is a depending on the botanical source and the geogra-serious health problem in dairy goats. Therefore, any phical origin of the beehive. The change in honey factor that affects the quantity and quality of goat milk composition is known to affect its antimicrobial and is of great financial interest. The inflammatory response antioxidant properties because of differences in levels was associated with a marked reduction of milk yield in of peroxide and non-peroxide factors [10]. This effect the infected gland (0.29 kg/milking) compared to that has been recently investigated on different types of of the uninfected [1]. Many mastitis pathogens were honey [6, 10-12]. considered as agents of major concern because of their The potent in vitro activity of honey against low response to the treatment and their propensity to antibiotic-resistant bacteria [13-15] and its successful recur chronically [2]. Targeted antimicrobial therapy application in treatment of infections is a promising plays an important role in mastitis control. It reduces research topic as confirmed by the published reports of the levels of herd infection and prevents new infection many researchers [7, 9, 11, 15-17]. Furthermore, the [2]. Now-a-days, the abundant use of antibiotics has large spectrum of the antibacterial activity of honey is resulted in widespread resistance. As far as mastitis is multi-factorial in nature. Indeed, hydrogen peroxide considered, it is important to know the type of and high osmolarity-honey consisting of 80% (w/v) of antibiotic to recommend for the treatment and the way sugars are the only characterized antibacterial factors it should be introduced (parenteral or intramammary). in honey [18]. Recently, high concentrations of Incorrect or incomplete treatment of infections may antibacterial compound consisting of methylglyoxal cause an antibiotic resistance. (MGO) were found [19, 20]. Honey has a valuable role in traditional medicine In the present study, we evaluated in vitro the worldwide for thousands of years. It is recognized as an antimicrobial drugs susceptibility of isolated bacterial efficient topical antimicrobial agent in the treatment of strains from subclinical mastitis goat's milk and then many infections [3, 4]. Recently, there is a renewed investigated the antimicrobial activity of locally interest in the study of honey as a natural product for …


Introduction
infected wounds [5][6][7][8][9].Note that honey has no unique composition but various types could be collected Mastitis inflammation of the mammary gland is a depending on the botanical source and the geograserious health problem in dairy goats.Therefore, any phical origin of the beehive.The change in honey factor that affects the quantity and quality of goat milk composition is known to affect its antimicrobial and is of great financial interest.The inflammatory response antioxidant properties because of differences in levels was associated with a marked reduction of milk yield in of peroxide and non-peroxide factors [10].This effect the infected gland (0.29 kg/milking) compared to that has been recently investigated on different types of of the uninfected [1].Many mastitis pathogens were honey [6, [10][11][12].considered as agents of major concern because of their The potent in vitro activity of honey against low response to the treatment and their propensity to antibiotic-resistant bacteria [13][14][15] and its successful recur chronically [2].Targeted antimicrobial therapy application in treatment of infections is a promising plays an important role in mastitis control.It reduces research topic as confirmed by the published reports of the levels of herd infection and prevents new infection many researchers [7,9,11,[15][16][17]. Furthermore, the [2].Now-a-days, the abundant use of antibiotics has large spectrum of the antibacterial activity of honey is resulted in widespread resistance.As far as mastitis is multi-factorial in nature.Indeed, hydrogen peroxide considered, it is important to know the type of and high osmolarity-honey consisting of 80% (w/v) of antibiotic to recommend for the treatment and the way sugars are the only characterized antibacterial factors it should be introduced (parenteral or intramammary).
in honey [18].Recently, high concentrations of Incorrect or incomplete treatment of infections may antibacterial compound consisting of methylglyoxal cause an antibiotic resistance.

Honey has a valuable role in traditional medicine
In the present study, we evaluated in vitro the worldwide for thousands of years.It is recognized as an antimicrobial drugs susceptibility of isolated bacterial efficient topical antimicrobial agent in the treatment of strains from subclinical mastitis goat's milk and then many infections [3,4].Recently, there is a renewed investigated the antimicrobial activity of locally interest in the study of honey as a natural product for collected Algerian honey against these isolated bacteria.therapeutic purposes including the treatment of

Bacterial strains and inoculum standardization:
The study was conducted on local crossbreed goat (Arabia x Makatia) in Tiaret province.The studied bacteria inoculated into Muller Hinton agar with the swab such were isolated from subclinical mastitis goats' milk.A that the whole surface of the agar is covered.total of 298 samples were collected during the year The antibiotic disks were dispensed on the 2010.The isolates were cultivated on various culture surface of the medium and incubated aerobically at a media including MacConkey agar, Chapman agar, temperature 37ºC during 24h.The results were recorded as either resistant or susceptible by measuring nutritive agar, and Columbia blood agar, at a the diameters (mm) of the inhibition zones according to temperature 37° C during a lapse time of 24 h.After the the interpretive standards of the NCCLS.purification of bacterial isolates, the selected colonies were subjected to the coloring of Gram finally galleries

Api (Api staph 20, Api E 20).
tration measurement: Honey sample was obtained The inoculum suspensions of tested strains were directly from beekeepers in the region of Tiaret, obtained by taking five colonies from 24 hour-old Algeria (350°15' N, 10°26' E).It was stored in dark at cultures grown on specific media.They were suspentemperature 4º C in tan containers before use.The ded in 5 ml of sterile saline (0.85% NaCl).The obtained evaluation of the antibacterial activity of honey inoculum suspensions were shaken for 15 seconds and dilutions were performed according to the method of the density was adjusted to a turbidity of 0.5 MacFarland Hegazi [6].To test the efficiency of honey against 6 Standard (equivalent to 1-5 x 10 CFU/ml).mastitis pathogen isolates varying concentrations of honey (10% -20%, v/v) were incorporated into the culture Antibiotics susceptibility test: Antibiotic susceptimedia [21][22][23].The final volume of honey and culture bility test was performed on the mostly isolated media in each plate was adjusted to a total of 5 ml.The pathogens (Coagulase Negative Staphylococci (CNS), plates were incubated at a temperature 37°C during 24 Pseudomonas aeruginosa, Streptococcus D; Enteroh.The minimum inhibitory concentration (MIC) was bacter spp., Escherichia coli, Klebsiella spp., Bacillus defined as the lowest concentration of honey inhibiting spp., Staphylococcus aureus, Corynebacterium spp.and the visible growth of each organism on the plate.Note Micrococcus spp.).The antibiotic assay was performed that all the MIC values are expressed in percentage using disk containing the following antibiotics (Ctra de (v/v).Santa Coloma, 7 17176 Sant Esteve de Bas, Girona, SPAIN): Penicillin-G (10IU); Ampicillin (10µg); Amoxicillin Statistical analysis: Calculations were performed th (25µg); Streptomycin (10µg); Erythromycin (15µg), using Statistica software (6 version).The data were and Tetracycline (30µg).

Hinton agar according to the National Committee of Clinical Laboratory Standards (NCCLS). The colonies
The antibiotic susceptibility rate of the isolated were suspended in 5 ml of sterile saline having a bacteria from clinically normal goats' milk is detailed density of approximately 0.5 MacFarland Standard.A in Table-1 (a, b, c, d).We noticed that both Micrococcus dry sterile cotton wool swab was placed in the spp and Klebsiella spp.showed a susceptibility to suspension and excess liquid was pressed against the Streptomycin and Tetracycline.Whereas P. inside of the tube.The bacterial suspension was aeruginosa, E. coli, Enterobacter spp., Bacillus spp.and CNS were preferably susceptible to Streptomycin, mastitis pathogens strains to antibiotics is of high Streptococcus D and Corynebacterium spp.were importance from both clinical and economic points of resistant.Indeed, Streptococcus D was the most view.Lactam antibiotics are frequently used in the resistant to the tested antibiotics whereas S aureus was treatment of mastitis in the goats [24].In the present the most susceptible to all the tested antibiotics work, most of the tested bacteria showed a resistance to (Figure -1).lactam antibiotics.Some CNS species have been The results regarding the antimicrobial activity of reported to be resistant against Penicillin G in caprine honey against the isolated bacteria are presented in [25] and bovine mastitis [26].This resistance may be Table-2.We notice that honey was effective within the due to the incorrect or incomplete antibiotics treatments [27].concentration range of 11-14% as it completly inhibits In the present study, the results obtained showed the growth of bacteria.The Algerian honey showed that S. aureus, Bacillus spp., Enterobacter, Klebsilla higher activity against Corynebacterium spp.(13%), spp.and Micrococcus spp.were susceptible to the most Pseudomonas aeruginosa (11.96±3.13%),CNS (12.66 tested antibiotics.Moroni et al. [24] and Tras et al. [27] ±4.27%), Klebsiella spp.(13.37±2.63%),Micrococcus have already reported similar susceptibility in the goat spp.(13.55±2.06%),Enterobacter spp.(13.09± 0.84%), mastitis.Moreover, susceptibility to lactams and E. coli (13.53±3.95), S. aureus (13.25±0.95%),fluroquinolones was reported in the ewes [28,29] and Streptococcus D (14 ± 1.22%) and Bacillus spp.(13 ± bovine mastitis [26].Streptococcus D and P. aeroginosa 4.24%).
were resistant to the most tested antibiotics in contrast

Discussion
to the results obtained by Aydin and collaborators [30], wherein Streptococcus spp. was found to be The study of the sensitivity or resistance of

Table - 1
(c): Antibiotic susceptibility of bacterial isolates ( from subclinical mastitis goats Micrococcus spp.and Klebsiella spp.) D Figure-1: Percentage of resistance of strains isolated from subclinical mastitis goats.

Table - 2
: Antimicrobial activity of honey against bacteria isolated from subclinical mastitis goat CNS: Coagulase Negative Staphylococci susceptible to Lactams, Erythromycin, Oxytetracyclin mastitis might be necessary before the treatment.The and Florfenicol.study also shows that Algerian honey exhibited in vitro In the current investigation, the results of antimicrobial activity against different bacteria isolated Minimum Inhibitory Concentration revealed that from goat mastitis.The honey could be a promising honey was effective against CNS, Staphylococcus aureus, alternative therapy in the treatment of infected udder.Streptococcus D, Corynebacterium spp., Klebsiella However, future investigations are needed to confirm spp., P. aeruginosa, Enterobacter spp., E. coli, and the in vivo antimicrobial activity of honey in the Bacillus spp.All values of MICs registered were treatment of subclinical mastitis in goat.between 10% and 14%.Many reports literature revealed