Introduction
Mastitis is a worldwide problem of dairy production characterized by physical and chemical changes in milk. The microbiological and pathological changes in the glandular tissue of the udder alter the quality of milk. Mastitis has emerged as a health hazard responsible for the economic losses in dairy animals (Cady et al., 1983). It has been noted that amongst all diseases, mastitis is most expensive and ranks at the highest point list of disease of dairy animals by various studies led in Pakistan (Khan et al., 1991). The losses due to mastitis could be even higher in Pakistan due to the non-adoption of the mastitis prevention practices (Arshad, 1999). Mastitis incidence in overall disease of dairy production is (38%) approximately (Kossaibati and Esslement, 1997). Mastitis leads to great economic losses in the form of poor production of milk, increased treatment cost, reduction of milk following treatment, culling chronic animals and labor expenditures (Miller et al., 1993). Culling the chronically infected buffaloes, curing clinical mastitis cases with effective antibiotic during lactation, dry cow therapy and spontaneous cure the dry period are other component of mastitis control programs (Philpot and Nickerson 1994). Broad spectrum antibiotics administered in the beginning of dry period prevent new IMI and significantly increased the cure rate of existing intramammary infection compared to lactation period (Davidson et al.,1994). Prevalence of mastitis is high in dairy cattle but low in buffalo because buffalo have strong teat canal as compared to cattle (Thapa and Kaphle, 2002).Dairy management systems especially bedding material plays an important role in control of mastitis. It was found that sand bedding was the best bedding material because it contains no nutrients and provides no harboring place to mastitis causing bacteria (Raza and Khan, 2000). Researchers have indicated that effective dry therapy enables the cure of 50 to 98 % intramammary infection (Batra 1988;Kirk 1991; Radostits; Leslie; Fetrow 1994). Dry cow therapy is practice of treatment, an intra-mammary infusion of antibiotic in udder or systemic treatment with an antibiotic administered at the time of drying or the end of lactation. Aims of antibiotic treatment at drying off are excluding the existing intra-mammary infections and avoiding new infections (Janosi and Huszenicaza, 2001). The route of intramammary antibiotic treatment has advantage of high absorption of antibiotic in udder region while demerits contain micro-organisms infection during inoculation and anatomical and physical injury to teat canal and teat cistern (Bradley et al., 2003). In a number of these studies beside intramammary therapy systemic and combined antibiotic therapy were administered during drying off. Tetracycline, sulpona-mide-trimetoprim, and floroquinolones were administered via the systemic route (Smith et al.,1985). The purpose of this study was to compare the efficacy of systemic and combined antibiotic therapies with intramammary administrations on the reduction of new infection and the cure of existing intramammary infection.
Materials and Methods
The animals used in this study were obtained from Livestock Experiment Station, University of Agriculture Faisalabad. Twelve Nili-Ravi buffaloes at the end lactation period were selected regarding the AI records, divided into four groups and, following administration, were done regardless of the laboratory results.
Group I. (n=3): one dose of dry products including Tylosin 30 mg/kg via intramuscular route were administered to all buffaloes.
Group II. (n=3): one tube of dry buffalo product including Gentamicin was administered into all of quarters.
Group III. (n=3): combinations of both above intramammary and intramuscular antibiotic administrations were used.
Group IV (n=3): No treatment applied to this group.
Collection of milk sample
Ten ml of milk sample from each quarter of buffaloes was collected aseptically before treatment drying off, and 14th day post completion of treatment. Collection of milk samples was done aseptically according to National Mastitis Council guidelines (Hogan et al. 1999). Each teat was scrubbed with cotton gauze socked with 70 percent alcohol. Immediately following milk sample collections and antibiotic was given to the animals. The collected samples were preserved at 40C and shifted to laboratory in the Institute of National Institute of Food Science and Technology for composition analysis of milk. Milk fat and protein was determined by Gerber method (Aggarwal and Sharma, 1961), and formal titration (Davide, 1977), respectively. Solid not fat, specific gravity and total solid were calculated by using the following formula as described by (Davide, 1977). Somatic cell count was calculated by milk film preparation for somatic cell count (Schalm et al., 1971). Lactose contents were estimated by Fehling’s solution titration method as described by (Egan et al., 1981).
Statistical analysis: The Data recorded was analyzed under Completely Randomized Design using computer software MINITAB (2000) and the mean was compared using the Tuckey’s test to draw the valid conclusion at certain significance level as described by (Steel et al., 1997).
Results
Before treatment at 0 day the prevalence rate of clinical mastitis to all mammary quarters and infected mammary quarters were shown in Table 1. The overall prevalence of clinical mastitis (CM) at the start (0 day) of experiment showed (58.33%) in all animals used for the experiment. The highest incidence (66.66%) of clinical mastitis was present in Group GII, GIII, GIV, but in Group G1 incidence of clinical mastitis (33.33%) was low as compared to other groups. The cure and prevention rate of clinical mastitis after treatment at 14th day were shown in Table 2. The cure rate on buffalo basis in Group I and Group II were (33.33%) while Group III where combined therapy given cure rate was (0%). In the control group cure rate was (66.66%). Analysis of the data on the basis of animal’s quarters subclinical mastitis at 0 day showed in Table 3. Out of forty eight (48) quarters only fourteen (14) quarters (29.16%) were affected for subclinical mastitis (SCM). It was noted that in control group G4 and group GI three quarters (25%) were showed subclinical mastitis. The Group G2 and Group G3 four quarter (33.33%) were affected by subclinical mastitis. After treatment at 14th day the cure and prevention rate of clinical mastitis was showed in Table 3. Overall prevalence of clinical mastitis was (33.33%) in all animals. The highest incidence of clinical mastitis (66.66%) was prevalent in control group (G4). The Group (G1) and (G2) receiving systemic and intramammary treatment (33.33%) incidence of clinical mastitis were recorded. It was observed that when animals received G3 treatment (both systemic and intramammary) no case of clinical mastitis (CM) was recorded. Analysis of the data on the basis of animal’s quarters subclinical mastitis at 14th day of treatment showed in Table 4. Out of forty eight (48) quarters only 5 (10.41%) were affected by subclinical mastitis. The control group (G4) had shown three quarters (25%) infected for subclinical mastitis. It was observed that when animals received G3 treatment therapy both (systemic and intramammary infusions) no case of clinical mastitis was recorded which indicated 100% cure rate within the group. In group G1 and G2 the recovery rate was (8.33%). The milk samples analyzed for Specific gravity, fat (F), protein, total solids (TS), and solid not fat (SNF) and lactose to investigate the effect of treatments on milk quality both at 0 day and at the14th days of therapy. Milk composition in different groups of Nili-Ravi buffaloes at 0 day showed in table 5. The highest Fat % was observed in G3 (5.67 ± 0.07) and the lowest (5.59 ± 0.11) found in G1 group. The value for G2 (5.60 ± 0.00) and G4 (5.63 ± 0.12) differed significantly. Total solids % in G2 (15.65 ± 0.09) and G3 (15.76 ± 0.12). Concentration of total solids were less in G1 (14.48 ± 0.19) as compared to both G2 (15.65 ± 0.09). Solid not fat concentration (8.89 ± 0.23) in group G1 and G2 (10.05 ± 0.15) while G3 (10.09 ± 0.18) and G4 (8.94 ± 0.28). In group G1 Protein concentration was (3.96 ± 0.05) and group G2 (4.30 ± 0.05) while in group G3 and G4 protein % was (4.30 ± 0.20) (3.84 ± 0.21). Lactose value in group G1 and G2 (3.90 ± 0.15), (4.44 ± 0.07) while in group G3 and G4 lactose value was (4.40 ± 0.05) (3.96 ± 0.03). Specific gravity value in group G1 and G2 Was (1.01 ± 0.05) (1.01 ± 0.05) while Group G3 and G4 Specific gravity value was (1.01 ± 0.20) (1.01 ± 0.05). Milk composition in different groups of Nili-Ravi buffaloes at 14th day showed in table 6. The highest Fat % (6.33 ± 0.03) was observed in G3 group and the lowest (5.63 ± 0.12) found in group G4 (control). While in group G1 and G2 fat % was (6.03 ± 0.14) (6.07 ± 0.14). The highest total solid % was observed in G3 (16.66 ± 0.31) and the lowest (14.58 ± 0.34) found in group G4. While in group G1 and G3 total solid % was (15.07 ± 0.215) (16.25 ± 0.32). The percentage of solid not fat in group G1 and G2 was (9.04 ± 0.25), (10.18 ± 0.19). While in group G3 and G4 solid not fat percentage was (10.43 ± 0.24), (8.94 ± 0.28). The protein percentage in group G1 and G2 was (3.92 ± 0.15), G2 (4.58 ± 0.12)While in group G3 and G4 protein percentage was (4.91 ± 0.11), (3.84 ± 0.21). Lactose value in group G1 and G2 was (4.29 ± 0.12), (4.63 ± 0.06)While in group G3 and G4 lactose value was (4.89 ± 0.05), (3.96 ± 0.03). Specific gravity value in group G1 and G2 was (1.018 ± 0.0018), (1.02 ± 0.00). While in group G3 and G4 specific gravity value was (1.02 ± 0.00), (1.01 ± 0.00). In the combined therapy group, all existing infection was cured (100%) as compared to other groups and also improved milk composition. In the control group, the existing infection remained the same while in systemic and intramammary groups the cure rate was (8.33%) respectively.
Discussion
In this study, apparently healthy animals were suffering from clinical and subclinical mastitis on analysis of milk samples in the laboratory before start of experiment. The uppermost prevalence of clinical mastitis (66.66%) was present in Group G2, G3, and G4 while in Group G1 incidence of clinical mastitis (33.33%) was low as compared to other groups. Moroni et al., (2006) described the prevalence of intramammary infection (IMI) in buffalo (66%) especially during dry period due to the environmental microorganisms. The prevalence of CM found at 0 day in this experiment was in line with the mentioned earlier workers. Zecconi et al., (2000) also described that the orthodox management practices of small buffalo holder in rural areas as the probable reason for increased percentage of subclinical mastitis in indigenous dairy animals. Out of the total (24) rear quarters 11 (45.83%) were affected by SCM, while in case of front udder only 3 (12.2%) cases of were observed for SCM. The findings of the present study are in agreement with the mentioned researchers. Maximum prevalence of SCM was found in rear quarters (45.83%). Bayazit and Izgur (2006). Analysis of data after 14th day of post treatment the combined therapy showed better results which eliminate the existing and upcoming infection. Similar study was conducted by Bayzait and Hakki (2006) reported that the combined therapy were better results which eradicate the prevailing and upcoming infection. Cure Rate of combined therapy were (100%) than systemic and intramammary therapy. The maximum recovery rate of subclinical mastitis (100%) was found in group G 3. The results of the present research are in agreement with Musal and Izgur (2006) that the rates of prevention of mastitis by combined therapy were higher than the sole systemic therapy.
Ullah et al., (2005) reported that the intensity of clinical and subclinical mastitis had an effect on milk composition and fat percentage. Hortet et al. (1998) also observed a slight reduction in milk fat %age in dairy cows due to mastitis. Barbano (1989) revealed that mastitis prevalence affects the milk composition in terms of decrease in milk protein, fat, lactose contents which supported the findings of this study. Malek et al. (2013) also reported that the reduction in milk constituents had been associated with the increase in somatic cell counts and severity of the tissue damage and impairment of the ability for milk synthesis. Haggag (1991) reported that specific gravity of milk depends upon the udder health status of the animal. In mastitis milk specific gravity reduced due to increased chlorides and decreased lactose contents. At 14th day of therapy the result showed that combine therapy cured and prevention rate is higher than systemic and intramammary therapy. Musal and Izgur (2006) reported that rates of elimination and prevention of intramammary infection and for intramammary and combined therapy group were higher than systemic therapy group. Regarding theses finding combine therapy systemic plus intramammary (Gentamicin+ Tylosin) can be used effectively to cure subclinical IMI inn dry period. The cure and prevention rates obtained by these combined drugs is higher than cure rates obtained by similar drugs.
Table 1: Animal wise prevalence of clinical mastitis before treatment at 0 day
Table 2: Animal wise prevalence of clinical mastitis at 14th day treatment
Table 3: Quarter wise prevalence of sub-clinical mastitis before treatment at 0 day
Table 4: Quarter wise prevalence of sub-clinical mastitis at 14th day of treatment
Table 5: Milk composition in different groups of Nili-Ravi buffaloes at 0 day
Table 6: Milk composition in different groups of Nili-Ravi buffaloes at 14thday
References
Aggarwala, A. C. and R. M. Sharma. 1961. A laboratory manual of milk inspection. Asia publishing House, Bombay, India.
Arshad, G. M. 1999. A population based active disease surveillance and drug trails of mastitis in cattle and buffaloes of District Sargodha. M.Sc. Thesis, Deptt. Vet. Clinical Medicine and Surgery, Univ. Agri., Faisalabad, Pakistan.
Batra, T. R. 1988. Effects of a complete dry cow treatment on mastitis control in dairy cattle. J Anim Sci. 68:553-556.
Bayazit, M. and H. Izgur. 2006. The efficacy of intramammary, systemic and combined antibiotics administered during dry off in cows with subclinical mastitis. Ankara Üni. Vet. Fak. Derg. 53:175-178.
Berry, E. A. and J. E. Hillerton. 2002. The effect of selective dry cow treatment on new intramammary infections. J. Dairy Sci. 85:112-121.
Bradley, A. J. 2002. Bovine mastitis. An evolving disease. Vet. J. 164:116-128.
Bradley, A. J. and J. N. Huxley. 2003. A rational approach to dry cow therapy in making logical treatment decisions. In. Pract. 25:12-17.
Cadey, R. A., S. K. Shah, E. C. Schermerhorn and R. E. Mcdowell. 1983. Factors affecting performance of Nili-Ravi buffaloes in Pakistan. J. Dairy Sci. 66:578-586.
Davide, C. L. 1977. Laboratory guide in the dairy chemistry practical. FAO regional dairy development and training center for Asia and the Pacific. Dairy training and research institute, University of Philippines Los Bamos College, Langnene.
Davidson, T. J., I. R. Dohoo, A. W. Donald. 1994. Comparing two dry cow treatments on the new infection and elimination rates of coagulase negative staphylococci. Vet J. 35:775 -776.
Egan, H., R. Kirk and R. Sawyer. 1981. Person chemical analysis of food. Churchill Livingstone, NY, USA.
Fetrow, j., S. Stewart, S. Eicker, R. Farnsworth and R. Bey. 2000. Mastitis: an economic consideration. National mastitis council annual meeting proceedings, Atlanta, GA. pp. 3-47.
Haggag, H. F., L. F. Hamzawi, G. A. Mahran and M. M. Ali. 1991. Physico chemical properties of colostrum and clinical and subclinical mastitic buffalo milk. Egyptian J. Dairy Sci. 19:55-63.
Hogan, J. S., R. N. Gonzalez, R. J. Harmon, S. C. Nickerson, S. P. Oliver, J. W. Pankey and K. L. Smith. 1999. Laboratory handbook on bovine mastitis. National mastitis council, Madison. pp. 6-10.
Hortet, P., H. Seegers, P. Hortet and H. Seegers. 1998. Loss in milk yield and related composition changes resulting from clinical mastitis in dairy cows. Prev. Vet. Med. 37:1-20.
Khan, M. A., M. Ajmal, M. Yamin, M. S. Khan and M. A. Athar. 1991. Epidemiological and economical based ranking order of buffalo and cattle diseases through active surveillance system. Pak. J. Livestock Res. 1:38-43.
Khan, M. Z. and A. Khan. 2006. Basic facts of mastitis in dairy animals. Pak. Vet. J. 26:204-208.
Kirk, J. H. 1991. Diagnosis and treatment of difficult mastitis cases, Staphylococcus and Pseudomonas. Agri-Pract. 12:5-8.
Kossaibati, M. A. and R. J. Esslemont. 1997. The cost of production diseases in dairy herds in England. Vet. J. 154:41-51.
Malek, D. R., J. R. Barreiro, L. Mestieri, M. A. Porcionato and M. V. Santos. 2013. Effect of somatic cell count and mastitis pathogens on milk composition in Gyr cows.BMC Vet. Res. 9:65-67.
Miller, G. Y., P. C. Barlet, S. E. Lance, J. Anderson and L. E. Heider. 1993. Cost of clinical mastitis and mastitis prevention in dairy herds. Amer. J. Vet. Med. Assoc. 202:1230-1236.
Musal, B. and I. H. Izgur. 2006. The efficacy of intramammary, systematic and combined antibiotics administered during dry off in cows with subclinical mastitis. Ankara Univ. Vet. Fak. Derg. 53:175-178.
Oliver, S. P., M. J. Lewis, B. E Gillespie, H. H. Dowlen, E. C. Jaenicke and R. K. Roberts. 2003. Prepartum antibiotic treatment of heifers: Milk production, milk quality and economic benefit. J. Dairy Sci. 86:1187-1193.
Philpot, W. N. and S. C. Nickerson. 1994. Mastitis Counter Attack, Babson Bros. Co., Illinois.
Radostits, O. M., K. E. Leslie, J. Fetrow. 1994. Mastitis control in dairy herds. Herd Health 2nd Ed. W.B. Saunders Company, Philadelphia. 229-273.
Raza, S. H. and S. khan. 2000. Effect of bedding on behavior and milk production in Nili Ravi buffaloes. ADSA and ASAS Joint meeting at Baltimore.
Schalm, O. W., E. J. Carol and N. C. Jain. 1971. Bovine mastitis Lea and Febiger, Philadelphia, USA.
Smith, K.L., D. A. Todhunter, P. S. Schoenberger. 1985. Environmental pathogens and intramammary infection during the dry period. J Dairy Sci. 68:402-417.
Steel, R. G. D., J. H. Torrie and D. A. Dicky. 1997. Principles and procedures of statistics, a biometric approach. 3rd edition, MC Graw Hill Book Co., New York, USA.
Thapa, B. B. and K. Kaphle. 2002. Selecting different drugs combination for the control of bovine mastitis. J. Ani. Vet. Adv. 1:18-19.
Uallah, S., T. Ahmad, M. Q. Bilal, G. Muhammad and S. U. Rahman. 2005. The effect of severity of mastitis on protein and fat contents of buffalo milk. Pak. Vet. J. 25:1-4