Effect of Non-Genetic Factors on Somatic Cell Count in Tharparkar Cows under Hot Arid Region of Thar Desert Area

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DOI: 10.5958/2277-940X.2016.00144.3 Journal of Animal Research: v.6 n.6, p. 1001-1005. December 2016 Effect of Non-Genetic Factors on Somatic Cell Count in Tharparkar Cows under Hot Arid Region of Thar Desert Area Amit Sharma 1*, Sanjita Sharma 1, Navav Singh 1, Rahul Singh Pal 2 and Brijesh Nanda 1 1 Department of Livestock Production Management; Post Graduate Institute of Veterinary Education and Research, Jaipur, Rajasthan, INDIA 2 Livestock Research Station, Chandan, Jaisalmer, Rajasthan, INDIA *Corresponding author: A Sharma; Email: amitkanota@gmail.com Received: 05 July, 2016 Accepted: 29 September, 2016 ABSTRACT The study was conducted to determine the effect of non genetic factors on Sub Clinical Mastitis (SCM) and to investigate the relationships between these factors with Somatic cell count (SCC) in Tharparkar Cattle raised at Livestock Research Center, Chandan, Jaisalmer in Thar desert of Rajasthan, India. Quarter wise milk samples were collected and examined by somatic cell count to check the status of subclinical mastitis. Data were collected from the record register maintained at farm and data evaluated by Parity, Stage of Lactation and Season of calving by the SPSS packet program. On analysis of variance result revealed that the parity had highly significantly (P<0.01) effect on SCC. Higher incidence of SCM was in later parities in comparison to primiparous animals. No statistical difference was found between different stages of lactation. Season of calving had significant (P<0.05) effect on SCC. Lower incidence of SCM and SCC were revealed in summer and winter season of calving as compared to the autumn and rainy season of calving groups. So it can be concluded from the present study that the non genetic factors like parity, stage of lactation and season of calving have a significant effect on Milk SCC. Thus, dairy farmers are advised to spend more time on their herds and given extra care to their animals in later parity stage, early stage of lactation and in the autumn or rainy season of calving to prevent the infection of sub clinical mastitis. Keywords: Somatic cell count, Subclinical mastitis, Parity, Stage of lactation, Season of calving Subclinical mastitis is economically one of the most costly and menacing disease of dairy industry worldwide and stands as the major obstacle towards healthy milk production (Bhartiet al., 2015). Among the several barriers in achieving the milk production targets, subclinical mastitis (SCM) continues to remain as a most challenging impediment, as the infected udder quarters show 4-18 per cent less production (Neilson et al., 2009). Somatic Cell Count is a helpful predictor of intra mammary infection (IMI), and therefore, an important component of milk in assessment of aspects of quality, hygiene and mastitis control. Somatic cells are mainly milk-secreting epithelial cells that have been shed from the lining of the gland and white blood cells that have entered the mammary gland in response to injury or infection (Dairyman s digest, 2009). Somatic cell count is influenced by several factors, including host and environmental factors. Housing and managemental practice are the major environmental factors while age, stage of lactation, parity and season of calving is the major host related factors. The most risk factors associated with management and the environment are addressed by introducing extra management during highest susceptibility, hygiene measures and selecting dairy cows, which are less susceptible to mastitis is also a control measure worthy of consideration. A number of studies have been done to investigate the relation of these non-genetic factors i.e. parity, stage of lactation, season, calving season etc. with occurrence of intramammary infection in exotic and cross bred cattle under different climatic conditions but information regarding Indigenous cattle breeds are scanty. So the present study

Sharma et al. was conducted to determine the effect of parity, stage of lactation and season of calving on milk Somatic Cell Count in Tharparkar cattle. obtain the number of somatic cells per ml of the milk. Due to SCC not displaying a normal distribution, data of SCC were log transformed to base 10. MATERIALS AND METHODS Location The study was conducted at the Livestock Research Station, Chandan (RAJUVAS), cattle yard located in Jaisalmer city of Rajasthan, India. The altitude of Jaisalmer city is 238.21 meters above mean sea level, latitude and longitude position being 26 54' 57N and 70 54' 51E, respectively. Animal Out of total number of animals in milk total, 105 healthy lactating Tharparkar cows with no evidence of clinical mastitis were selected and the evaluation of udder and teat traits and the collection and analysis of milk samples were performed on the same day. Collection of samples and diagnosis of Intra-mammary Infection Representative milk samples were collected from all the four quarters of lactating cows. For this purpose, quarters were designated as Left Fore (LF), Left Hind (LH), Right Fore (RF) and Right Hind (RH). About 30 ml of milk was collected aseptically in the clean sampling bottles after discarding the first 2-3 streaks of fore milk. The collected samples were brought to the laboratory immediately for further analysis. Within 6 h of collection, the milk samples were spread on 2 microscope slide areas, which were 10X10 mm 2 in size subsequently. The slides were fixed with pouring of ethyl alcohol for 2 minutes and the prepared smears were stained with the modified Newman s Lampert stain, by keeping the prepared slide in the staining solution for 1 to 2 minutes. The smears were gently washed in tap water and dried. The dried stained smears were examined under the oil immersion lens of the microscope. Thirty different fields per smear were observed, and the average number of somatic cells per field was calculated. The average number of cells per field was then multiplied by the microscopic factor of the microscope, i.e. 393174 to Data Collection Data were collected from the record register maintained at farm and data of SCC from all animals were classified according to parity, stage of lactation and season of calving. All these parameters were taken into consideration as for the factors influencing the occurrence of SCM and SCC. Parity: All animals were classified under the different parities (viz., 1 st, 2 nd, 3 rd, 4 th, 5 th and above) according to their lactation number. Stage of lactation: The whole lactation was partitioned early stage of lactation (up to 90 days), mid stage of lactation (91 to 180 days) and late stage of lactation (181 days & above). Season of calving: All the animals were classified in to four calving seasons on the basis of their date of calving. Cattle which calved during december to february fall under winter, march to may under summer season, june to august under rainy season and cows which calved during september to november month fall under autumn season of calving categories. Statistical analysis Obtained SCC values were transformed to log 10 SCC for normality and homogeneity of variance. In the study, parity, stage of lactation and season of calving were evaluated as independent variables. The data were examined by Analysis of Variance (ANOVA) and means were compared by Duncan s multiple range test. The model was as follows: Where, y ijklm = µ + a i + b j + c k + d ijklm y ijklm = Observation value for SCC µ = Population mean a i = Effect of Parity (I = 1,2,3,4, 5 and above) b j = Effect of Stage of lactation (j =1,2,3) c k = Effect of Season of calving (k = Summer, Rainy, Winter and Autumn) d ijklm = The Random residual effect 1002 Journal of Animal Research: v.6 n.6 December 2016

Effect of Non-Genetic Factors on somatic cell count All statistical analysis was performed using SPSS software statistical package (16.0). RESULTS AND DISCUSSION Results of SCC in milk samples of healthy cows are presented in Table 1. The overall animal wise and quarter wise prevalence on the basis of SCC was 52.38 (55/105) and 40.80% (162/397), respectively. The arithmetic mean ± SE of SCC (absolute and logarithmic) of animal wise milk samples were 220472 ± 37189.50 cells/ml of milk and 5.343 ± 0.037 respectively. Range of SCC varied from 39,370 cells/ml of milk in healthy quarter to 1916007 cells/ml of milk in severely infected quarters. Table 1: Incidence, Mean ± SE values & range of somatic cell counts Number of observations Per cent Incidence of SCM 105 52.38 Mean ± SE (cells/ ml) 220472 ± 37189.50 SE=Standard error, SCC=Somatic cell count Effect of Parity on SCC Mean ± SE ( SCC) 5.343 ± 0.037 Range (cells/ ml) 39370-1916007 After statical analysis results of the present study revealed that incidence of SCM increased with number of parity. The lowest incidence was observed in 1 st parity (19.54 per cent), gradually increased and highest in >5 th parity (70.00 per cent) in Tharparkar cows. Several workers reported similar findings that are enhancing milk production level and a rise in SCC with Increasing parity in cows and buffaloes (Sharma, 2007; Sharma and Prasad, 2002; Kavitha et al., 2009; Saravanan et al., 2015). The analysis of variance showed that the effect of parity on SCC was highly significant (P<0.01). Mean SCC of Individual parity was compared by Duncan s Multiple Range Test. The results of post hoc test revealed that the mean SCC of 1 st, 2 nd and 5 th parity differed significantly (P<0.05) from each other. The difference between the means SCC of 3 rd parity was nonsignificantly differ from 1 st and 2 nd parity group, whereas the means SCC of 4 th parity was non-significantly differ from 2 nd and 5 th parity of animals. Similar significant association of parity with log 10 SCC were reported by Erden et al. (2010), Yu et al. (2011), Houda et al. (2012) and Saravanan et al. (2015) whereas Singh and Ludri, (2001), Ahn et al. (2006) and De et al. (2011) reported non-significant effect of parity on SCC. The higher incidence of SCM and high SCC in later parity in comparison to primiparous animals may be due to the fact that constant exposure of animals to the pathogens and loosening of sphincters, help the easy entry of organism through streak canal which is responsible for higher rates of infection in multiparous animals in comparison to their younger counterparts. Further, animal s resistance to mastitis might also be lowered with the advancement of parity or age. Yu et al. (2011) reported that with the increase of parity, the ratio of macrophages (MAC) and polymorphonuclears (PMN) tended to be increased and suggested that progressive increase of SCC when parity increased. Heuven et al. (1988) claimed the increase of SCC over parities shows that older cows were more likely to suffer prolonged elevation of SCC or even prolonged udder damage. Table 2: Mean ± SE of SCC in different non genetic factors in Tharparkar cows Traits Number of observations Mean ± SE of SCC Parity 397 5.384±0.021 1 87 5.191±0.037 a 2 152 5.366±0.033 b 3 87 5.377±0.043 ab 4 21 5.570±0.080 bc 5 and Above 50 5.704±0.065 c Stage of Lactation 397 5.384± 0.021 Early stage 113 5.371±0.030 Mid stage 165 5.361±0.032 Late stage 119 5.428±0.036 Season of calving 397 5.384±0.021 Summer season 228 5.342± 0.032 a Rainy season 49 5.476±0.045 ab Autumn season 23 5.597±0.053 c Winter season 97 5.386±0.030 b Mean showing different superscripts in lower case letters in respective categories in a column differ significantly (p<0.05), SE=Standard error, SCC=Somatic cell count Journal of Animal Research: v.6 n.6 December 2016 1003

Sharma et al. Streak canal of the teats consists keratinized layer and accumulated cellular debris, which is wax like and having sebum like nature, consisting of long chain fatty acids, which provides bacteriostatic effects (Mostert et al., 2001). The breakdown of such streak canal barrier with advancing lactation leads to increased frequency of infection with successive increase in parity. The incidence of SCM and SCC increased as the lactation progressed and it attained the highest proportion during the 5 th and above lactation. Table 3: Analysis of variance showing the effect of non genetic factors on SCC in Tharparkar cows Source of variance Degree of freedom Sum Square Mean Sum square F value Parity 4 9.137 2.284 14.099** Error (Parity) 392 63.515 0.162 Stage of Lactation 2.334 0.167 0.910 Error (Stage of Lactation) 394 72.319 0.184 Season of Calving 3 1.870 0.623 3.461* Error (Season of calving) 393 70.782 0.180 SCC=Somatic cell count, ** Highly Significant (p<0.01), *Significant (p<0.05) Effect of Stage of lactation on SCC The prevalence of SCM in Tharparkar cows were 38.93, 38.78 and 45.38 per cent and mean ± SE values of SCC has been 5.371±0.030, 5.361±0.032 and 5.428±0.036 in early, mid and late stages of lactation, respectively. Results reported higher incidence of SCM and high SCC in early stage followed by slightly decrease in mid lactation stage and highest in late stage of lactation. The analysis of variance revealed that the stage of lactation had a non significant effect on the SCC (Table 3). The Duncan multiple range test revealed that the difference between the mean SCC of early, mid and late stage of lactation were not differ significantly from each other. Ceron- Munoz (2002), Ghoshet al. (2004), Ahnet al. (2006) and Sharma et al. (2007), found higher incidence of SCM and high SCC in early and late stage of lactation. Ceron- Munoz et al. (2002) reported that SCC in uninfected cows is high at freshening, lowest from peak to mid-lactation, and highest at drying off. Harmon (1994) suggested that a modest rise in the SCC of uninfected quarters at the end of lactation is in fact a dilution effect. Dahoo and Meek (1982) reported that the SCC increases with progressing lactation (late lactation) regardless of whether the cow is infected or not. Saravanan et al. (2015), Houda et al. (2012), De et al. (2011) and Wicks and leaver (2006) reported non-significant changes in SCC during different stages of lactation which support the results of the present study. Effect of Season of calving on milk SCC The per cent quarter-wise incidence of SCM was 37.72, 46.97, 69.57 and 38.15 per cent in summer, rainy, autumn and winter season of calving respectively. The mean (±SE) SCC in case of summer, rainy, autumn and winter season of calving was 5.342 ± 0.032, 5.476 ± 0.045, 5.597 ± 0.053 and 5.386 ± 0.030 respectively. The analysis of variance revealed that the season of calving has significant (P<0.05) effect on SCC. The mean SCC of winter, summer and autumn calving cows were significantly differ from each other, whereas mean SCC of rainy season calving was non-significantly differ from summer and winter calving cows but significantly differ from autumn season calving cows. Baul et al. (2011) and Wicks and Leaver (2006) supported the present findings and reported that the season of calving had a significant effect (P<0.01) on changes in the somatic cell count. Lower incidence of SCM and SCC in summer and winter season of calving as compared to the autumn and rainy season of calving can be better explained by the adaptability of animals to the environment (Reis et al., 2013). Tharparkar cows are native breed of Thar region which are more adoptable in hot and arid region. Environmental conditions of hot arid region are humid during the rainy and autumn season, which favor the growth of microorganism. Animals calved during autumn and rainy season are more susceptible to infection, having a greater number of somatic cells count in raw milk. It can be concluded from the present study that the nongenetic factors, i.e. parity, season of calving and Stage of lactation have some degree of association with prevalence of subclinical mastitis. Thus, dairy farmers are advised 1004 Journal of Animal Research: v.6 n.6 December 2016

Effect of Non-Genetic Factors on somatic cell count to spend more time on their herds and given extra care to their animals in later parity, first stage of lactation and in autumn or rainy season of calving which may help to reduce the incidence of mastitis in indigenous cattle. ACKNOWLEDGMENTS We are grateful to Dean PGIVER, Jaipur (RAJUVAS) for providing facilities, continuous encouragement and support during the study. REFERENCES Ahn, B.S., Suh, G.H. and Kwon, E.G. 2006. Estimation of environmental effects and genetic parameters for somatic cell score, stress and immunological traits in Holstein cattle. J. Anim. Sci. Technol., 48: 9-14. Baul, S., Cziszter, L.T., Acatincai, S., Cismas, T., Gavojdian, D., Tripon, I., Erina, S. and Raducan, G. 2011. Researches regarding the season influence on somatic cell count in milk during lactation in Romanian black and white cows. Lucrari Stiintifi ce., 56: 16 Bharti, P., Bhakat, C., Ghosh, M.K., Dutta, T.K. and Das, R. 2015. Relationship among intramammary infection and raw milk parameters in jersey crossbred cows under hot-humid climate. J. Anim. Res., 5: 317-320. Ceron-Munoz, M., Tonhati, H., Duarte, J., Oliveira, J., Munoz- Berrocal, M. and Jurado-Gamez, H. 2002. Factor affecting somatic cell count and their relations with milk and milk constituent yield in buffaloes. J. Dairy Sci., 85: 2885-2889. Dahoo, I.R. and Meek, A.H. 1982. Somatic cell counts in bovine milk. Can. Vety. J., 23: 119. Dairyman s Digest, 2009. What you should know about somatic cells. Winter issue, 2010. www.medvet.umontreal.ca/rcrmb. De, U.K. and Mukherjee R. 2009. Prevalence of mastitis in crossbred cows. Indian Vet. J., 86: 858-859. Erden, H., Atasever, S. and Kul, E. 2010. A study on somatic cell count of jersey cows. Asian J. Anim. Vet. Adv., 5: 253-259. Ghosh, C.P., Nagpaul, P.K. and Prasad, S. 2004. Factors affecting sub clinical mastitis in sahiwal cows. Indian J. Dairy Sci., 57: 127-131. Harmon, R.J. 2001. Somatic cell counts: A primer. In: Proc. National Mastitis Council Annual Meeting, pp. 3-9. Heuven, H.C.M., Bovenhuis, H. and Politiek, R.D. 1988. Inheritance of somatic cell count and its genetic relationship with milk yield in different parities. Livest. Prod. Sci., 18: 115 127. Houda, H., Trujillo, A.J., Juan, B., Guamis, B., Eifeki, A. and Gargouri, A. 2012. Interrelationships between somatic cell counts, lactation stage and lactation number and their influence on plasmin activity and protein fraction distribution in dromedary (Camelus dromedaries) and cow milks. Small Rumin. Res., 105: 300 307. Kavitha, K.L., Rajesh K., Suresh, K., Satheesh K. and Sundar, N.S. 2009. Buffalo mastitis risk factors. Buffalo Bull., 28: 135-137. Mostert, B.E., Theron, H.E. and Kanfer, F.H.J. 2001. The effect of calving season and age at calving on production traits of South African dairy cattle. South Afr. J. Anim. Sci., 31: 205-214. Nielsen, C.H., Emanuelson, U., Berglund, B. and Strandberg, E., 2009. Relationship between somatic cell count and milk yield in different stages of lactation. J. Dairy Sci., 92: 3124-3133. Reis, C.B.M., Barreiro, J.R., Mestieri, L., Porcionato, M.A.F. and Santos, M.V. 2013. Effect of somatic cell count and mastitis pathogen on milk composition in Gyr cows. BMC Vet. Res., 9: 1-7. Samanta, A., Prasad, S. and Ghosh, C.P. 2006. Incidence of sub clinical mastitis in Karan swiss and Karan Fries crossbred cows. Indian J. Dairy Sci., 59: 13-18. Saravanan, R., Das, D.N., De, S. and Panneerselvam, S. 2015. Effect of season and parity on somatic cell count across zebu and crossbred cattle population. Indian J. Anim. Res., 49: 383-387. Sharma, N. 2007. Alternative approach to control intramammary infection in dairy cows- A review. Asian J. Anim. Vet. Adv., 2: 50-62. Singh, M. and Ludri R.S. 2001. Influence of lactation, parity and season on somatic cell counts in cows. Asian-Aust. J. Anim. Sci., 14: 1775-1780. Wicks, H.C.F. and Leaver, J.D. 2006. Influence of genetic merit and environment on somatic cell counts of Holstein Friesian cows. Vet. J., 172: 52 57. Yu, A.B., Zhao, G.Q., Tian, S.Q. and Huo, Y.J., 2011. Relationship between parity and cellular composition of somatic cells in milk of Chinese Holstein cows. J. Anim. Vet. Adv., 10: 2067-2073. Journal of Animal Research: v.6 n.6 December 2016 1005