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4World's
Veterinary Journal
WVJ
World's Vet. J. 2(2): 13-16, 2012
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Influence of Staphylococcus aureus Mastitis on Milk Composition of Different Dairy Breeds of Cattle in Khartoum State, Sudan
I.M.B. Hamid1, E.S. Shuiep1,2, I.E.M. El Zubeir1*, A.Z. Saad3, and O.A.O. El Owni1
1Department of Dairy Production, Faculty of Animal Production, University of Khartoum, Khartoum North Postal code 13314 Sudan 2Department of Animal Production, Faculty of Veterinary Science, University of Nyala, Nyala, Sudan 3Department of Microbiology, Faculty of Veterinary Medicine, University of Khartoum, Khartoum North, Sudan
The purpose of the present study was to investigate the effect of S. aureus mastitis on the compositional quality of milk. Sixty milk samples were collected from 19 Frisian, 16 cross and 25 local cows suffering from subclinical (14) or clinical (46) mastitis from different farms in Khartoum State, Sudan. Total bacterial count, S. aureus count and some compositional quality were estimated. Non-significant (P>0.05) differences were recorded between the milk obtained from the three groups of dairy cattle for total bacterial count. However, significantly (P<0.05) higher S. aureus count was obtained (2.7* 103± 0.7 cfu/ ml) for milk samples from cross- bred cows. Chemical analysis revealed no significant (P>0.05) differences in total solids (11.87+ 1.7 and 11.1± 1.6%), protein (4.3± 2.8 and 3.6± 0.5%), fat (3.1± 0.7% and 3.10± 0.5), lactose (2.3+ 0.5 and 2.1± 0.7%) and acidity (0.211± 0.14 and 0.44± 0.36 %) for milk obtained from cows with subclinical and clinical mastitis, respectively. However, significantly higher ash content was found in milk samples collected from clinical mastitis (0.68± 0.12%) compared to subclinical ones (0.6 + 0.15%). Milk constituents of infected milk by S. aureus revealed non-significant (P>0.05) differences expect for lactose (P<0.01), which showed variations between breeds. Total bacterial count showed significantly (P<0.05) negative correlation (r= -0.037) when compared with lactose content of mastitic milk. It could be concluded that mastitis caused by S. aureus would resulted in the decrease the nutritive content of milk.
KEY WORDS: Staphylococcus aureus, Mastitis, Milk Composition, Dairy Breeds, Sudan
INTRODUCTION
Mastitis caused by Staphylococcus aureus is recognized worldwide in dairy cows as subclinical and clinical intramammary infection (Akineden et al., 2001); it is of considerable importance in Sudan (Mohamed et al., 1997; El Zubeir et al., 2006). The infection is spread at milking time when S. aureus contaminated milk from an infected gland comes in contact with an uninfected gland, and the bacteria penetrate the teat canal. One reason for the failure to eradicate S. aureus intramammary infection may be the control measures for mastitis do little to control this disease in prepartum period in heifers (Roberson et al., 1998). Moreover Ebliny et al. (2001) reported that S. aureus causes infection of longer duration.
Recently, it was reported that 3% of all animals are infected with S. aureus (Schukken, et al., 2009). Tenhagen et al. (2009) added that S. aureus infection represents 10 to 12 % of clinical mastitis cases. Moreover, in a study conducted in Egypt, it was found that 16% of all mastitis cases are caused by S. aureus (Hameed et al., 2006). In Ethiopia, examination of 300 lactating cows (140) 46.7% mastitis cases; 10.0% and 36.7% for clinical and subclinical infections, respectively (Abera et al., 2010). Moreover, with severe clinical mastitis, abnormalities of milk are easily observed and such milk is discarded by the producer. However, milk of cows with subclinical mastitis, (with no visible changes) is accidentally mixed into bulk milk; it enters food chain and can be dangerous to humans (Hameed et al., 2006). Contagious mastitis, transmitted from cow to cow during milking is mainly caused by S. aureus (Fox and Gay, 1993).
The onset of udder infection (mastitis) is known to influence the chemical composition of milk constituents (Mohamed et al., 1997). Staphylococcus aureus causes severe tissue damage that result in excretion of cells into milk (Kitchen, 1981; (Petersson-Wolfe et al., 2010). Clinical mastitis infection detected by California Mastitis Test (CMT) has a significant effect on the total solid of milk (Mohamed et al., 1999), Bovine S. aureus strains express many
_| Hamid Intisar M.B., Shuiep E.S., El Zubeir I.E.M., Saad, A.Z. and El Owni, O.A.O. 2012. Influence of Staphylococcus Aureus Mastitis on Milk
Composition of Different Dairy Breeds of Cattle in Khartoum State, Sudan. World's Vet. J. 2(2): 13-16. | | http://wvj .science-line.com/
Corresponding author's email: [email protected]
ABSTRACT
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virulence factors, which may enable this organism to cause chronic infection (Sutra and Poutrel, 1994). Further Middleton (2002) reported that S. aureus caused a significant (P<0.01) decrease in the infected quarter compared with the uninfected quarters in the same cow. Hence the objectives of the study were isolation, enumeration of S. aureus and assessment of its influence on composition of milk with clinical and subclinical mastitis along with breed dependent differences in the innate immune response in different cow breeds to S. aureus intramammary infection
MATERIALS AND METHODS
Animal
In the present study, Friesian, local breeds (Kenana and Butana) and cross breed (local cows and Friesian) from farms in Khartoum State were examined for presence of clinical and subclinical mastitis. Mastitis milk samples were collected from Shambat (n=18), Tayba farm (n=4), Azaheer Company farm (n=8), El Radwan Project (n=9), Western Elgrif area (n=5), El Ailafoon (n=6), Eid Babikr (n=6) and Hamza farm (n=4).
Collection of milk samples
The udder was first cleaned with soap then dried with clean towels. The teats were swabbed with 70% alcohol. Visual observations and (CMT) were done before the collection of samples to group the cows into clinical and subclinical cases. Then, approximately 100 ml milk from each cow was taken into sterile Mc-Cartney bottles.
Microbiological examination
Total bacterial count was done on plate count agar (Merck, Darmstadt, Germany) by pour plating methods following the manufactures instructions (Marshall, 1992). The appropriate dilution of milk samples was selected and the plates were incubated at 32° C for 48 hours. For the determination of S. aureus, Mannitol salt agar medium (Oxoid, Hampshire, England) was used followed by confirmation by biochemical test. The plate counting 25- 250 colonies were selected as described by Houghtby et al. (1992). All counts were done in duplicates, multiplied by the reciprocal of the dilution and reported as colony forming unit per milliliter (cfu/ ml).
The presence of S. aureus
Staphylococcus aureus isolates were Gram stained and subjected to motility and catalase test, acid production and oxidation fermentation (OF) test. Moreover, fermentation of sugars, methyl red (MR), Voges-Proskauer (VP) and tube coagulase test were done as secondary confirmatory tests according to the criteria outlined by Barrow and Feltham (1993).
Chemical analysis
The chemical constituents including total solids, fat (Gerber's method), protein (Kjeldahl method) and ash of milk samples were done according to AOAC (1990). The lactose was determined by anthrone method according to Richards (1959). The acidity of milk samples was also determined using the titration methods according to AOAC (1990).
Statistical analysis
Data were arranged into computer coding format to facilitate the statistical analysis. The SPSS (Statistical Package for Social Science) was used by using randomized complete block design to differentiate between the intensity of infection and different breeds of cows. Also the graphs was plotted using excel program.
RESULTS AND DISCUSSION
The numbers of mastitic cows detected from Friesian, local and cross breed in Khartoum State are shown in Figure 1. The isolation of S. aureus from subclinical and clinical mastitic cows indicated that S. aureus is one of important pathogens causing mastitis. This result was in line with Tenhagen et al. (2009). Moreover, in dairy cows, S. aureus is considered a contagious organism, which easily spread between cows during milking (Paape et al., 2000). Staphylococcus aureus was isolated in Sudan from medical laboratories environment by El Mansouri (1997), from mastitic cows' milk by Mohamed et al. (1997) and El Zubeir et al (2006) and from raw camels' milk by Shuiep et al. (2009).
Figure1. Comparison of prevalence of S. aureus mastitis in different cows at dairy farms in Khartoum State
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Differences were reported between the three cattle breeds for the total bacterial count (Table 1). Milk from Friesian cows showed less S. aureus count than local cows. This result supported Bannerman et al. (2008) who reported that S. aureus intramammary infection showed differences in the innate immune response in different cow breeds. Those could be attributed to resistance of local cows to bacterial infection (El Deeb and Hassan, 1987). On the other hands, Berry et al. (2007) reported that Jersey cows have lower prevalence of mastitis than Friesian cows. In all 3 types of cows the total bacterial count was more compared to S. aureus count (Table 1).
Table 1. Chemical composition and microbial counts of milk from mastitic dairy cows in Khartoum State
Measurements Friesian Cross breed Local breed Sig. level
(n=19) (n=16) (n=25) (P-value)
Acidity 0.28±0.3 0.33±0.3 0.35±0.3 0.86 NS
Total solids 11.3±1.6 10.95±1.8 11.66±1.5 0.50 NS
Ash 0.68±0.13 0.7±0.14 0.62±0.12 0.23 NS
Fat 3.2±0.41 2.99±0.6 3.1±0.68 0.61 NS
Protein 3.4±0.55 3.66±0.35 4.3±0.2 0.26 NS
Lactose 2.42±0.6 1.67±0.7 2.3±0.6 0.009 **
S. aureus count (cfu/ml) 5.9X102±0.9 2.7X103±0.7 1.5x103±0.5 0.05*
Total bacterial counts (cfu/ml) 1.04X104±0.6 1.9X105±0.9 1.04x103±0.12 0.47 NS
N.S = Non Significant; * = Significant (P < 0.05); ** = = Significant (P < 0.01).
Table 2. Comparison between chemical composition and microbial counts of subclinical and clinical mastitic milk of
dairy cow in Khartoum State
Measurements Subclinical Clinical Sig. level
(P-value)
Acidity (%) 0.211±0.14 0.44±0.36 0.2 NS
Total solids (%) 11.87±1.7 11.1±0.6 0.19 NS
Ash (%) 0.6±0.15 0.68±0.12 0.05 *
Fat (%) 3.1±0.7 3.1±0.54 0.93 NS
Protein (%) 4.3±2.8 3.6±0.5 0.16 NS
Lactose (%) 2.3±0.5 2.1±0.7 0.38 NS
S. aureus count (cfu/ml) 6.3X102±0.9 1.9x103±0.7 0.21 NS
Total bacterial counts (cfu/ml) 7.9X104±0.2 1.4x105±0.2 0.4 NS
N.S = Non Significant; * = Significant (P < 0.05); ** = Significant (P < 0.01).
The present study indicated variations in means of bacterial counts and milk constituents for subclinical and clinical mastitic milk samples (Table 2). The mean bacterial count for clinical mastitis milk (1.9* 103+ 0.7 cfu/ ml) was higher than that of subclinical mastitis milk (6.3*102±0.9 cfu/ ml). This was in accordance with the finding of El Deeb and Hassan (1987) who reported that total bacterial count increased when milk tested positive for mastitis. Also Mohammed et al. (1997) reported high log bacterial count for clinical followed by subclinical mastitis milk compared to those from healthy cows. It also agrees with the findings of Bautista et al. (1986) who reported a level of 104- 105 cfu/ ml S. aureus in milk from subclinical infected ewes. The levels of total protein in this study (Table 1 and Table 2) were found to be similar to those reported by Kitchen (1981) who concluded that total protein content in mastitic milk does not change with increasing the level of infections in the mammary gland.
Table 2 showed that the mean acidity of clinical mastitic milk is higher than that of subclinical mastitis milk. Mastitic milk from Friesian, cross and local cows revealed mean acidity of 0.28+ 0.3%, 0.33+ 0.3% and 0.35±0.30%, respectively (Table 1). The lower values of titratable acidity obtained during the present study is in agreement with those of Schulz et al. (1988) who reported significant association between mastitis and low milk acidity, which might be due to more alkaline constituents of the blood that passing into the milk. The lactose value of the present study showed lower values when compared with those of healthy cows, which supports Mohamed et al. (1997). Moreover, mastitic milk was reported to have lower lactose contents (Gunha, 1989), especially in the staphylococcal infection (Mohamed et al., 1997).
The relatively high significant level (P<0.01) of ash observed during the present study when comparing subclinical cases to clinical ones was in agreement with the finding of El Deeb and Hassan (1987). The fat content from mastitis milk was found to be lower than those from healthy cows. The reduction of fat may be due to the increase rate of lipolysis during infection (Murphy et al., 1989).
It is concluded that the compositional content of milk from mastitic animals showed variations that influences by the breed and the degree of infection. Moreover in subclincal mastitis, when the farmers were unable to recognize the disease public health hazards might occur due to consumption of infected milk that contains pathogenic bacteria or their toxins.
ACKNOWLEDGEMENT
The financial support received from Ministry of Higher Education and Scientific Research is acknowledged with thanks. The authors also appreciate the cooperation of the farm owners.
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REFERENCES
Abera, M, Demie1 B, Aragaw K, Regassa F and Regassa A, 2010. Isolation and identification of Staphylococcus aureus from bovine mastitis milk and their drug resistance patterns in Adama town, Ethiopia. Journal of Veterinary Medicine and Animal Health, 2: 29- 34.
Akineden, O, Annemuller C, Hassan AA, Lammler C, Wolter W and Zschock M, 2001. Toxin genes and other characteristics of Staphylococcus aureus isolates from milk of cows with mastitis. Clinical and Diagnostic Laboratory Immunology, 8: 659- 964.
AOAC, 1990. Official Methods of Analysis, 15th ed., Association of Official Analytical Chemists, Washington, DC. USA.
Barrow, GI and Feltham RKA, 1993. Manual for the Identification of Medical Bacteria. 3rd, Cambirdge. UK.
Bannerman, DD, Springer HR, Paape MJ, Kauf CW and Goff JP, 2008. Evaluation of breed dependent differences in the innate immune responses of Holstein and Jersey cows to Staphylococcus aureus intramammary infection. Journal of Dairy Research, 75: 291- 301.
Bautista L, Bermejo MP and Nunez MI, 1986. Seasonal variation and characterization of micrococaceae present in ewe's raw milk. Journal of Dairy Science, 53, 1- 5.
Berry, DP, Lee JM, Macdonald KA, Stafford K, Matthews L and Roche JR, 2007. Associations among body condition score, body weight, somatic cell count, and clinical mastitis in seasonally calving dairy cattle. Journal of Dairy Science, 90: 637- 648.
Ebliny, LT, Fox LK, Bayles KW, Bohach GA, Byrne KM, Davis WC, Ferens WA and Hillers, JK 2001. Bovine mammary response to an experimental intramammary infection with a Staphylococcus aureus strain containing a gene for Staphylococcal enterotoxin C1. Journal of Dairy Science, 84: 2044- 2050.
El Deeb, SA and Hassan HN, 1987. Changes in the cow's milk composition as affected with mastitis infection. Alexandria. Journal of Agricultural Research, 32: 163- 174.
El Mansouri HA, 1997. Studies on the prevalence of saprophytic staphylococci in medical laboratories. Ph. D. V. Sc., Thesis. University of Khartoum, Sudan
El Zubeir IEM, Kutzer P and El Owni OAO, 2006. Frequencies and antibiotic susceptibility patterns of bacteria causing mastitis among cows and their environment in Khartoum State. Research Journal of Microbiology, 1: 101- 109.
Fox IK and Gay JM, 1993. Contagious mastitis. Vet. Clin. North. Am. FoodAnim. Pract., 9: 475- 487.
Gunha MD, 1989. Contribution to the clinical diagnosis of mastitis: Influence of phase of lactation milk phase inflammatory processes on physiochemical cellular and microbiological composition of the milk of the Holstein Friesian breed. Cited from Vet. Bull., 59: 1364.
Hameed, KGA, Sender G and Korwin-Kossakowska A, 2006. Public health hazard due to mastitis in dairy cows. Animal Science Papers and Reports. 25: 73- 85.
Houghtby, AG, Maturin LJ and Koenig KE, 1992. Microbiological Count Methods. In: Marshal, R.T. ed.: Standard Methods for the Examination of Dairy Products. 16th ed., American Public Health Association, Washington, D.C. USA.
Kitchen, BJ, 1981. Review of the progress of dairy science: bovine mastitis: milk compositional changes and related diagnostic tests. Journal of Diary Research, 48: 167- 188.
Marshall TR, 1992. Standard Methods for the Examination of Dairy Products. 16th ed. American Public Health Association, Washington, D.C. USA.
Middleton JR, LK Fox and TH Smith, 2001. Management strategies to decrease the prevalence of mastitis caused by one strain of Staphylococcus aureus in a dairy herd. JAVMA 218, 1615-1618.
Mohamed, IE, El Owni OAO and Mohamed GE, 1997. Effect of bacteria causing mastitis on milk constituents. Sudan Journal of Veterinary Science and Animal Husbandry, 36: 125- 136.
Mohamed, IE, El Owni OAO and Mohamed GE,1999. Effect of mastitis on milk quality of Friesian cows in Khartoum State. Indian Journal of Animal Sciences, 69: 733- 735.
Murphy, SC, Cranker K, Senyk GF, Brabano DM, Saeman AI and Gaiton DM, 1989. Influence of bovine mastitis on liypolysis in milk. Journal of Dairy Science, 72: 620- 626.
Paape, JM, Duenas MI, Wettemann RP and Douglass LW, 2000. Effects of intramammary infection and parity on calf weaning weight and milk quality in beef cows. Journal of Animal Science, 78: 2508- 2514.
Petersson-Wolfe, CS, Mullarky IK and Jones GM, 2010. Staphylocuccus aureus mastitis: cause, detection and control. Virginia Cooparative Extension, 404: 229.
Richards, EL, 1959. The reaction of lactose with anthrone and its application to the estimation of lactose in casein and the dairy products. Journal of Dairy Research, 26: 53- 57.
Roberson, J.R, Fox LK, Hancock DD, Gay JM and Besser TE, 1998. Source of intramammary infection from Staphylococcus aureus in dairy heifers at first parturition. Journal of Dairy Science, 81: 687- 693.
Schulz, J, Sterba P and Beck K , 1988. Changes in the acidity of raw milk with acute mastitis. Dairy Science Abstract, 50: 2616.
Shuiep, ES, Kanbar T, Eissa N, Alber J, Lämmler C, Zschöck M, El Zubeir IEM and Weiss R, 2009. Phenotypic and genotypic characterization of Staphylococcus aureus isolated from raw camel milk samples. Research in Veterinary Science, 86: 211- 215.
Sutra, L, Poutrel B, 1994. Virulence factor involved in the pathogenesis of bovine intramammary infection due to Staphylococcus aureus. Journal of Medical Microbiology, 40: 79- 89.
Tenhagen, B.A, Hansen I, Reinecke A and Heuwieser W, 2009. Prevalence of pathogens in milk samples of dairy cows with clinical mastitis and in heifers at first parturition. Journal of Dairy Research, 76: 179- 87.
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