The present research work was carried out to evaluate prophylactic potential of Tri-sodium citrate powder in subclinical mastitis in milking cows. The study involved twelve HF cross dairy cows in which each diagnosed with at least one subclinical mastitis quarter. The six (n=6) cows from conventional and six (n=6) cows from loose housing system with raised SCC were administered Tri-sodium citrate powder @ 30 mg/kg B.W. orally BID for five days. The SCC values of infected quarters were reduced significantly (P < 0.05) on 28th day after Tri-sodium citrate therapy as compared to value on day ‘O’ before therapy both in loose housing and conventional housing system. The percent efficacy in improvement of infected quarter after administration of Tri-sodium citrate powder in conventional and loose housing system was 63.63 % and 58.33 % respectively. Thus, results indicated that tri-sodium citrate having prophylactic potential in controlling mastitis in cows in both conventional and loose housing system.
Mastitis is one of the costliest diseases resulting in huge economic losses throughout the world including in India. Mastitis is characterized by a range of physical and chemical changes in the milk and pathological changes in the glandular tissue. The most important changes in the milk include discoloration, the presence of clots and the presence of large number of leukocytes (Constable et al., 2017). Mastitis is mainly divided by two categories: clinical and subclinical mastitis on the basis of clinical signs. Sub-clinical mastitis is a type of mastitis which is characterized by the no visual changes in the quarters and milk or the existence of inflammation in the absence of gross signs (Sharma et al., 2012). The economic losses due to bovine subclinical mastitis is in range of rupees 21,677/- to 88,340/- per animal per lactation (Rathod et al., 2017). Sub-clinically infected quarters can develop clinical mastitis and the rate of new infections can be high (Zdunczyk et al., 2003). Therefore, subclinical mastitis is one of the costliest and is alarming disease of dairy animals throughout the world and a major obstacle in milk production.
Citrate is the main constituent, playing a buffer role in the udder. It regulates the homeostasis between Ca++ and H+ ions for maintaining the fluidity of milk through its effect on casein micelles (Shennan and Peaker, 2000). In case of citrate deficiency in udder, the clumping of Ca++ appears which causes injury to the parenchymatous tissues of alveoli of udder. This leads to subsequent events such as the damage of barriers between blood and milk, and inflammatory reactions in the alveolar tissue of udder (Poutrel, 1982). Generally, mastitic milk has been found to be alkaline and low in citrate (Dhillon et al., 1989; Dhillon et al., 1995; Prakash et al., 2013). Tri-sodium citrate is safe, economical and very effective which avoid culling of animal and discard of milk. Also, it does not require withdrawal period because of residual problems in milk and meat (Dhillon and Singh, 2013). Tri sodium citrate may be useful for improving milk quality regarding somatic cell counts (Mbonwanayo et al., 2016). It was hypothesized that replacement of citrate deficiency with tri-sodium citrate might play some protective role against subclinical mastitis. Therefore, the present research was carried out to evaluate the prophylactic potential of tri-sodium citrate in crossbred cows under conventional and loose housing system.
Sarfaraz et al. (2009) studied evaluation of tri-sodium citrate alone and in combination with levamisole HCl in the treatment of sub-clinical mastitis and found that oral administration of tri-sodium citrate and levamisole HCl are viable alternative to antibiotic therapy for sub-clinical mastitis. Dhillon and Singh (2013) studied effect of tri-sodium citrate against clinical mastitis. The animals showed good recovery and citrate content of milk returned to almost normal levels after treatment. Further, the treatment with tri-sodium citrate proved safe, economical, very effective, with no discarding of milk, no withdrawal periods and no hazard from residuals in milk and meat. Reddy et al. (2017) reported that supplementation of coated tri sodium citrate @ 20 gms for 10 days in cows having sub-clinical mastitis had reduced pathogenic load, increased milk fat and milk production significantly. Santoshi et al. (2018) observed that supplementation of tri-sodium citrate during the transition period lowered SCC, pH and EC while improved productivity of indigenous dairy cows.
Material and Methods
Selection of Animals
In the present research project twenty-four HF crossbred lactating cows from conventional (n=12) and loose (n=12) housing systems in early to middle lactation having average weight around 400 kg were selected. In the selected cows six cows each from conventional and loose housing system were healthy while six cows from conventional and six cows from loose housing system were having at least one quarter positive for subclinical mastitis which were diagnosed by somatic cell count (SCC), modified california mastitis test (MCMT), pH and electrical conductivity (EC). The selected cows were randomly divided into four groups each having six cows (n=6). Group I and group III comprise of cows from conventional housing healthy control and loose housing healthy control, respectively which were maintained on regular diet; Group II and group IV were made up of cows from conventional housing system and loose housing system, respectively which were administered tri-sodium citrate powder @ 30 mg/kg B.W., orally BID for five days
Sampling and Diagnosis
About 5-10 ml of milk sample was collected from individual quarter of each cow in sterilized bottles and was subjected to various diagnostic tests viz. MCMT, pH and EC. The SCC test was performed in the laboratory within 1-2 hours after collection. The SSC has more accuracy to discriminate healthy milk from sub-clinical mastitic milk (Tiwari et al., 2018).The milk samples were tested to assess the quarter health status and milk samples were tested on day ‘0’ before institution of prophylactic regimen and on day 7, 14 and 28 after therapy in each group, for comparative evaluation, during the routine morning milking hours.
The data obtained in the current research work were statistically analyzed and compared by employing mean, standard error and CRD using WASP (Web Agri Stats Package) version 2.0.
Results and Discussion
The efficacy was recorded after observing the results of somatic cell count (SCC) modified California mastitis test (MCMT), pH and electric conductivity (EC) tests as shown in Table 1, 2 & 3. Therapy of tri-sodium citrate powder showed a significant (P<0.05) reduction in SCC, MCMT, pH and EC, in both conventional and loose housing system as compared with respective control and on day ‘0’ pre-treatment to day ‘28’ post-treatment.
Citrate is one of the main constituents of milk which plays a major role in buffering in the udder. Deficiency of citrate in udder is result into the clumping of Ca++ which results into injury to parenchymatous tissues in the udder alveoli. It means subsequent events such as the damage of barriers between blood and milk and inflammatory reactions would be established (Poutrel, 1982). Tri-sodium citrate has buffering and antimicrobial properties which can maintain normal pH of milk and subsequently prevent the formation of calcium ion crystals leads to cure from mastitis and improvement in milk quality (Shennan and Peaker, 2000; Yousaf et al., 2010). Tri sodium citrate may be useful for improving milk quality regarding somatic cell counts (Mbonwanayo et al., 2016).
Table 1: Prophylactic effect of Tri-sodium citrate on reduction in SCC
Table 2: Prophylactic effect of Tri-sodium Citrate on reduction in pH
Table 3: Prophylactic effect of Tri-sodium Citrate on reduction in EC
In current research work, the treatment group showed lower milk SCC, pH and EC levels as compared to the respective control group. This may be due to the citrate which plays a buffering role inside the mammary glands, it regulates the homeostasis between Ca++ and H+ ions and supports the fluidity of milk through its effect on casein micelles (Shennan and Peaker, 2000). Moreover, restoration of tri-sodium citrate in animals suffering from mastitis decreases pH of milk and perhaps acts against Ca++ injury and reduces the inflammatory process in the quarters of the udder, which in turn makes mammary secretary cells more efficient and ultimately the milk yield is increased (Prakash et al., 2013). Similar findings were reported by Sarfaraz et al. (2009), Rai et al. (2013), Dhillon and Singh (2013), Reddy et al. (2017), Prakash et al. (2010), Prakash et al. (2013), Santoshi et al. (2018), Pal et al. (1995) and Dhillon et al. (1995) for use of tri-sodium citrate for prevention and treatment of bovine mastitis.
The prophylactic effect on reduction of udder inflammation and percent efficacy of treatment in various groups is depicted in Table 4 and Fig. 1.
Table 4: Efficacy of therapy of tri-sodium citrate in conventional and loose housing system
|Group||Housing System||Prophylactic Therapy||Infected Quarters||Percent Efficacy|
|0 Day||7 Day||14 Day||28 Day|
Fig. 1: Therapeutic efficacy of tri-sodium citrate in conventional and loose housing system
The per cent efficacy of group II and IV is 63.63 and 58.33, respectively. This indicates that therapy of tri-sodium citrate could eliminate intramammary infection effectively. Therefore, it is clear that effect of tri-sodium citrate is slightly better in conventional housing system than in loose housing system.
Therapy of tri-sodium citrate powder @ 30 mg/kg B.W. orally twice a day for five days was found more efficacious in conventional housing system (63.63 %) than in loose housing system (58.33 %) in the prevention of subclinical mastitis in cows
I would like to express my special thanks of gratitude to my post gradation guide (Dr. S.U. Digraskar) as well as co-guide (Dr. M.F.M.F. Siddiqui). They gave me the golden opportunity to do this wonderful work which helped me in doing a lot of research and I came to know about so many new things related to mastitis. Secondly, I would also like to thank my parents and friends who helped me a lot in finalizing this project within the limited time frame.
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