A herbal tool- AV/KPC/10 to check the sub clinical ketosis in cows

 1. Introduction

During calving nutritional requirements increase rapidly with lactation. Ketosis, or acetonaemia, is a multifactorial disorder of energy metabolism i.e. metabolic disorder and is largely a management influenced disease in dairy cows in early lactation (Shpigel et al., 1996; Ingvartsen, 2006). Hyperketonemia is associated with an increase of oxidative stress levels (Youssef et al., 2010). Most of the animals in developing countries are fed on agriculture by-products and low quality crop residues have low nutritive value, i.e. low contents of metabolizable energy and crude protein (Vahora et al., 2013). Normally ruminants have low levels of blood glucose and during heavy milking when glucose utilization is high, even slight fall is enough to put the animal in a hypoglycemic state (Dufflied, 2006). During the early post partum period, milk production increases dramatically, while energy intake may not be adequate to sustain the higher production level. This results in negative energy balance and cows metabolize fat to meet their energy needs (Weber et al., 2012). During early lactation incidence rates of ketosis between about 40% and 60% (Duffield et al., 1998).

Nowroozi Asl et al., (2011) studied that 97 percent of the cows experienced at least one episode of subclinical ketosis during the sixth week post partum period there was a statistical relation between the prevalence of sub clinical ketosis and milk production. Ketosis leads to heavy economic losses. Geishauser et al. (2011) estimated cost of sub clinical ketosis per case was $78 CAD. A study by Varga (2004) summarized ketosis costs at $140 for treatment, and the total cost at $2,520 per year for a 120-cow farm. Economic losses are considerable due to decreased milk production, cost of treatment and occasionally from deaths or disposal of affected cows. The wide ranges of treatments have been recommended for ketosis. Most of these are intended to alleviate the hypoglycemia and increase the glycogen content of liver, thereby, restoring the carbohydrate metabolism and aims to provide an energy boost to overcome negative energy balance for optimizing the milk production.

The current study was designed to evaluate the efficacy of herbal gel AV/KPC/10 (M/S Ayurvet Limited) in the treatment of sub clinical ketosis in cows.

Total 87 lactating cows in early mid lactation were screened for ketosis by Modified Rothera’s test (Kelly, 1974) and Keto-Diastix Strip test in urine. 18 animals positive for sub clinical ketosis were selected and divided randomly into three equal groups, comprising of 6 cows in each group. Group I, cows positive for sub clinical ketosis were treated with AV/KPC/10 @ 200 gm twice a day orally for first two days followed by 100 gm once a day for next 3 days. Group II, cows positive for sub clinical ketosis were treated with Liquid Anabolite @ 200ml twice a day orally for first two days, followed by 100 ml twice a day for 3 day. Group III, positive for sub clinical ketosis were treated with Inj. Dextrose 25% @ 500 ml intravenously once in a day for three days. Total 12 healthy cows negative for sub clinical ketosis were selected and divided randomly into two equal groups, comprising of 6 cows in each group. Control Group IV, cows negative for sub clinical ketosis were kept without any treatment and treatment Group V, cows negative for sub clinical ketosis were supplemented with AV/KPC/10 @ 200 gm twice a day orally for first two days followed by 100 gm once a day for next 3 days. Biochemical parameters viz. plasma glucose (mg/dl), serum triglyceride (mg/dl), serum calcium (mg/dl), serum phosphorus (mg/dl) were determined on ‘0’ day (before treatment) and on 3rd, 7th and 30th day post treatment. The milk yield was recorded before treatment (‘0’ day) and daily after treatment during the experimental period.

The data collected during the present study with respect to different parameters was analyzed statistically by two ways factorial design using web based agricultural statistics software package (WASP), ICAR Research Complex, Ela, Old Goa.

In Group I, Group II and Group III cows were positive for sub clinical ketosis on day ‘0’ (before treatment) diagnosed by Modified Rothra’s Test and Keto-Diastix strip test.

In Modified Rothra’s Test, a mixture of reagent was taken up to half of the test tube. Urine sample was added slowly through the wall of test tube. The positive test showed purple colour formation in urine. The results are recorded as follows.

In Keto-Diastix strip test, dipstick containing the salt nitroprusside, which provide semi quantitative results. This test is based on the development of colours ranging from buff pink for a negative reading to purple when acetoacetic acid reacts with nitroprusside. The colour intensity varies with the amount of acetoacetic acid present in urine. Read the ketone test area by comparing with the colour chart at exactly 40 seconds after wetting and recorded the results. The Keto-Diastix® diagnostic test can be read in five categories: 1) negative (0 mg/dl), 2) trace (5 mg/dl), 3) small (15 mg/dl), 4) moderate (40 mg/dl), and 5) large (greater than 80 mg/dl of AcAc).

After treatment with AV/KPC/10, Anabolite and Dextrose 25% injection all the cows in respective groups were found negative for sub clinical ketosis on day 7th, 14th and 30th day. In Control Group IV, initially on day 0 all the cows were negative for sub clinical ketosis. During subsequent period 4 animals were found positive for sub clinical ketosis on 30th day. This means in untreated group 75% new cases of ketosis were observed. In Group V all the animals were negative for ketosis on day ‘0’ and no incidence of ketosis was observed on the day 3rd, 7th and 30th day after AV/KPC/10 supplementation. Which means AV/KPC/10 was efficacious to control the new incidences of ketosis.

 

 

In ketotic cows during early lactation period glucose supply is not sufficient for milk production (Padilla et al., 2005). Group I treated with AV/KPC/10 showed significant (P<0.05)improvement in plasma glucose level in comparison to groups treated with liquid Anaboliteand Injection Dextrose 25%. In AV/KPC/10 treated group I cows, on day 3rd (53.85 mg/dl) a significant (P<0.05) increase in plasma glucose level was observed in comparison of day 0 (43.05 mg/dl) and level was maintained till 30th day. The overall mean plasma glucose level (mg/dl) in AV/KPC/10 treated group (I) was 51.71 mg/dl. Increase in plasma glucose concentration is well in comparison with other supplemented groups. In Anabolite treatment group (II) and dextrose injection treatment (III) group, the overall mean plasma glucose values were 50.51 mg/dl and 49.94 mg/dl respectively (table 1). When treating cows for negative energy balance, it is essential to meet the need for glucose so that the ketogenic process in the liver are reversed (Sezer et al., 2010). The improvement in plasma glucose level was because the supplementation may have facilitated cellular uptake of glucose, suppresses fatty acid mobilization and stimulate hepatic gluconeogenesis (Radostits et al., 2000).

Table 1: Mean values of plasma glucose (mg/dl) in different groups before treatment and at different intervals after Treatment

Similar superscript shows non significant differences

The analysis of variances indicated significant (P<0.05) rise in plasma glucose on 3rd (52.68 mg/dl), 7th (57.77 mg/dl), and 30th (56.38 mg/dl) day (mean, 55.56 mg/dl) post AV/KPC/10 supplementation in group V (table 2). AV/KPC/10 supplementation has significantly maintained the plasma glucose concentration. In control group IV i.e. unsupplemented group there was a decrease in plasma glucose levels from 3rd (54.98), 7th (53.70) and 30th (49.00) day (Mean 53.64 mg/dl).

Table 2: Mean values of plasma glucose (mg/dl) in different groups of healthy animals before supplementation and at different intervals after supplementation

Similar superscript shows non significant differences

A significant (P<0.05) decrease in serum triglycerides levels was observed in all treatment groups from day 0 to day 30th. In group I, the triglycerides level was 40.37 mg/dl on day 0 which was significantly reduced to 34.12 mg/dl on day 3rd after AV/KPC/10 treatment and decreasing trend continued till day 7th (29.28 mg/dl) and 30th (26.18 mg/dl). The overall mean serum triglyceride levels (mg/dl) in group I treated with AV/KPC/10 was 32.49 mg/dl and was in well comparison with group II treated with liquid Anabolite (29.29 mg/dl) and group III treated with Injection.

Dextrose 25% was 31.57 mg/dl (table 3). Overall finding indicated that all the treatment found equally effective in improving triglyceride level in sub clinical ketotic cows within the experimental period.

Significant (P<0.05) reduction in serum triglyceride levels was observed in AV/KPC/10 supplemented Group V (pooled mean, 22.77 mg/dl) in comparison untreated group IV (pooled mean, 25.06 mg/dl) (table 4). The overall results indicated the effectiveness of AV/KPC/10 in restoring serum triglyceride level within treatment regimen.

Table 4: Mean values of serum triglyceride (mg/dl) in different groups of ketotic cows before treatment and at different intervals after treatment

Similar superscript shows non significant differences

Several workers reported, low level of serum calcium in sub clinical ketotic cows, which might be due to increase loss of base in urine to compensate ketosis induced acidosis (Duffield, 2006). The lower level of serum calcium level in subclinical ketosis can also be attributed to high concentration of β-hydroxybutarate (BHBA), which impairs the absorption and utilization of calcium in dairy cows during the early lactation period (Zhang et al., 2009).

The mean values of serum Calcium (mg/dl) in Group I, group II, group III and group IV of cowson day ‘0’ (before treatment) and on 3rd, 7th and 30th day after treatment are presented in Table 5. Serum calcium level was improved in AV/KPC/10 supplemented group I from day 0 (7.85 mg/dl) to day 3rd (7.95 mg/dl) and improvement was also observed on day 7th (8.23 mg/dl) till day 30th (8.32 mg/dl). Although overall result indicates that the non significant increase in serum calcium level was at par in animals treated with AV/KPC/10, Liquid Anabolite and Inj Dextrose 25% but the supplement products maintained the serum calcium level. Even in cows negative for ketosis i.e. in group IV and V, the mean serum calcium level varied non significantly (P<0.05) (table 6).

Table 5: Mean values of serum calcium (mg/dl) in different groups of cows before treatment and at different intervals after Treatment

Similar superscript shows non significant differences

Table 6: Mean values of serum calcium level (mg/dl) in different groups of healthy cows before supplementation and at different intervals after supplementation.

Similar superscript shows non significant differences

An important hydrogen buffering system involves phosphorus functions in the blood and is a critical component of nucleic acids, adenosine triphosphate (ATP), and adenosine monophosphate (AMP) (Cunningham, 2002). The lower level of serum phosphorus in ketosis might be due to compensation of ketosis induced acidosis, which triggers excretion of phosphorus via urine (Fatur, 1994).

Table 7: Mean values of serum phosphorus (mg/dl) in different groups of ketotic cows before treatment and at different intervals after treatment

Similar superscript shows non significant differences.

Serum Phosphorus level was improved significantly (P<0.05) in AV/KPC/10 supplemented group from day 0 (5.18 mg/dl) to day 30th (5.42 mg/dl) (table 7). Although statistical analysis revealed non significant variation (P<0.05) in serum phosphorus levels between all groups. According to Rollin et al., (2010) phosphorus concentrations did not differ between treatment groups even after injectable butaphosphan and cyanocobalamin.

Serum Phosphorus concentration significantly (P<0.05) improved in ketosis negative AV/KPC/10 supplemented group V cows (5.54 mg/dl) in comparison to ketosis negative unsupplemented group IV cows (5.31 mg/dl).

Table 8: Mean values of serum phosphorus level (mg/dl) in healthy cows before supplementation and at different intervals after supplementation

Similar superscript shows non significant differences

Milk yield is negatively correlated with acetone concentration in milk (Miettinen, 1994). Subclinical ketosis in early lactation reduces milk production; the average total loss in milk yield per cow has significant impact on economy (Rajala- Schultz et al., 1999; McArt et al., 2011, Paavo et al., 1993). The average milk yield (Kg) in Group I, group II, group III and group IV of cows on day ‘0’ (before treatment) and on 3rd, 7th and 30th day after treatment are presented in Table 9.

In group I treated with AV/KPC/10, the milk yield increased by 12.25% (4.58 Kg) on day 3rd, 17.1% (4.78 Kg) on day 7th, and 24.26% (5.07 Kg) on day 30th from day 0 (4.08 Kg). In group II the increase in milk yield was 30.91% (5.42 Kg) and in group III increase in milk yield was 19.52% (5.02 Kg) from day 0 to 30th. On other hand in unsupplemented group IV milk yield decreased by 2.69% (4.82 Kg) on day 0 to day 30th (4.69 Kg). Overall the pooled mean showed no significant (P<0.05) variation between various treatment groups.

Table 9: Average Milk yield (Kg) in different groups of cows at different intervals

Similar superscript shows non significant differences

A 9.18 % increase in milk yield was observed in AV/KPC/10 supplemented group V from day 0 (4.90 Kg) to day 30th (5.35 Kg) (table 10). The statistical analysis pooled mean revealed significant (p< 0.05) overall 7.9 % increase (P<0.05) in milk yield in AV/KPC/10 supplemented group V (5.19 Kg) as compared to normal control group IV (4.81 Kg).

Table 10: Average Milk yield (Kg) in different groups of cows at different intervals before and after supplementation.

Similar superscript shows non significant differences

The group of ketotic cows treated with AV/KPC/10 showed improvement in altered levels of plasma glucose, serum triglycerides and serum phosphorus on day 3rd till day 30th post treatment. All AV/KPC/10 supplemented cows were found negative for sub clinical ketosis on day 3rd, 7th and day 30th after supplementation. A significant increase in milk yield was observed after AV/KPC/10 supplementation. AV/KPC/10 supplementation was in well comparison with other treatments.

The authors are thankful to Ayurvet Limited, Baddi, India for providing necessary samples and guidance and Post Graduate Institute of Veterinary and Animal Sciences, MAFSU, Akola, for providing the required facilities, guidance and support. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.

1. Cunningham J.G., 2002. Textbook of Veterinary Physiology. 3rd ed. W.B. Saunders, Philadelphia, PA.

2. Duffield T.F., 2006. Minimizing Subclinical Metabolic Diseases in Dairy Cows. WCDS Advances in Dairy Technology 18, 43-55.

3. Dufflied T.F., 2006. Epidemiology of subclinical production diseases in dairy cows with an emphasis on ketosis. In: Proc. of 12th Intl. Conf. on Production diseases in farm animals by Joshi, N. and Herdt, T.H., 126-135.

4. Duffield T.F., Sandals D., Leslie K.E., Lissemore K., McBride B.W., Lumsden J. H. et al 1998. Efficacy of monensin for the prevention of subclinical ketosis in lactating dairy cows. Journal of Dairy Science 81, 2866-2873.

5. Fatur B., 1994. Ca, P, Mg, K and Na serum levels of dairy cows with subclinical ketosis. Proceedings of the 18th world buiatrics congress. 26th congress of the Italian association of buiatrics; Bologna: Societa'Italiana di buiatria, 1257-1260.

6. Geishauser T., K. Leslie, Kelton D., Duffield T., 2001. Monitoring for subclinical ketosis in dairy herds. Compendium on Continuing Education for the Practising Veterinarian 23, 65-71.

7. Ingvartsen K.L., 2006. Feeding- and managementrelated diseases in the transition cow: physiological adaptations around calving and strategies to reduce feeding-related diseases. Animal Feed Science and Technology, 126, (3-4), 175-213.

8. Kelly W.R., 1974. Veterinary clinical diagnosis. 2d ed. Baltimore: Williams and Wilkins, viii, 374 p.

9. McArt J.A., Nydam D.V., Ospina P.A., Oetzel G.R., 2011. A field trial on the effect of propylene glycol on milk yield and resolution of ketosis in fresh cows diagnosed with subclinical ketosis. Journal of Dairy Science 94 (12), 6011-6020.

10. Miettinen P.V., 1994. Relationship between milk acetone and milk yield in individual cows. Zentralbl Veterinarmed A. 41(2), 102-109.

11. Nowroozi Asl, A., Nazifi S., Ghasrodashti A.R., Olyaee A., 2011. Prevalence of Subclinical Ketosis in Dairy Cattle in the South Western Iran and Detection of cut off Point for NEFA and Glucose Concentrations for Diagnosis of Subclinical Ketosis. Preventive Veterinary Medicine 100, 38- 43.

12. Paavo V.A., Miettinen, Setälä J.J., 1993. Relationships between subclinical ketosis, milk production and fertility in Finnish dairy cattle. Preventive Veterinary Medicine 17 (1-2), 1-8.

13. Padilla L., Shibano K., Inoue J., Matsui T., Yano H., 2005. Plasma vitamin C concentration is not related to the incidence of ketosis in dairy cows during the early lactation period. Journal of Veterinary Medical Science 67(9), 883-886.

14. Radostits O.M., Gay C.C., Blood D.G., Linchilff K.W., 2000. Ketosis in ruminats. 9th ed. Veterinary Medicine: A Textbook of the Diseases of Cattle, Sheep, Pigs, Goats & horses. Saunders company, London. pp 1452-1462,

15. Rajala-Schultz P.J., Gröhn Y.T., McCulloch C.E., 1999. Effects of milk fever, ketosis, and lameness on milk yield in dairy cows. Journal of Dairy Science 82(2), 288-294.

16. Rollin E., Berghaus R.D., Rapnicki P., Godden S.M., Overton M.W 2010. The effect of injectable butaphosphan and cyanocobalamin on postpartum serum β-hydroxybutyrate, calcium, and phosphorus concentrations in dairy cattle. Journal of Dairy Science 93(3), 978-987.

17. Sezer K., Sahinduran S., Albay M.K., 2010. Investigation of effect of different treatment regimens on blood acid-base balance in cows with ketosis. Journal of animal and veterinary advances 9(2), 287-291.

18. Shpigel N.Y., Chen R., Avidar Y., Bogin E., 1996. Use of corticosteroids alone or combined with glucose to treat ketosis in dairy cows. Journal of the American Veterinary Medical Association 208(10), 1702-1704.

19. Vahora S.G., Parnerkar S., Kore K.B., 2013. Productive efficiency of lactating buffaloes fed bypass fat under field conditions: effect on milk  yield, milk composition, body weight and economics. Iranian Journal of Applied Animal Science 3(1), 53-58.

20. Varga G.A., 2004. Feeding strategies for transitioning cows into lactation. Proceedings of penn state dairy cattle nutrition workshop, 57-63.

21. Weber C., Hametner C., Tuchscherer A., Losand B., Kanitz E., Otten W., et al 2012. Variation in fat mobilization during early lactation differently affects feed intake, body condition, and lipid and glucose metabolism in high-yielding dairy cows. 2012. Journal of Dairy Science 96, 165-180.

22. Youssef M.A., El-Khodery S.A., El-deeb W.M., Abou El-Amaiem W.E., 2010. Ketosis in buffalo (Bubalus bubalis): clinical findings and the associated oxidative stress level. Tropical Animal Health and Production 42(8), 1771-1777.

23. Zhang Z., Liu G., Li X., Wang Z., Kong T., Zhang N et al., 2009. β-hydroxybutyrate, glucose, calcium, phosphorus, and vitamin C concentrations in blood of dairy cows with subclinical ketosis during the early lactation. Bulletin of the Veterinary Institute Pulawy 53, 71-74.