Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 55 issue 11 (november 2021) : 1271-1278

Monitoring Plasma Metabolic Profile, Fertility in Normal and Estrus Synchronized Suckled Kankrej Cows

Mehrajuddin Naikoo2, A.J. Dhami1,*, B.C. Parmar1
1Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand-388 001, Gujarat, India.
2Department of Gynaecology, Faculty of Veterinary Science and Animal Husbandry, SKUAST-Kashmir-190 006, Jammu and Kashmir, India.
Cite article:- Naikoo Mehrajuddin, Dhami A.J., Parmar B.C. (2021). Monitoring Plasma Metabolic Profile, Fertility in Normal and Estrus Synchronized Suckled Kankrej Cows . Indian Journal of Animal Research. 55(11): 1271-1278. doi: 10.18805/IJAR.B-4220.

ABSTRACT

Background: Postpartum fertility in dairy bovines is one of the important criteria of sustainable dairy industry and economy of farmers. The Kankrej, a dual purpose zebu cattle, is known for its endurance, but is slow breeder with very late maturity, prolonged postpartum anestrous/subestrous and thereby calving interval. These peculiarities necessitate scientists to adopt fixed time AI (FTAI) protocols of estrus synchronization for improving their reproductive efficiency. Hence this study was aimed to assess the fertility and plasma metabolic profile of normal and synchronized anestrous/ subestrous suckled Kankrej cows of an organized farm.

Methods: The study involved 36 parturient Kankrej cows of the University Farm that were maintained under identical nutritional and managerial practices. Blood samples were collected in heparinized vacutainers from all the animals on the day of calving and then at 10 days interval till 90 days for plasma metabolic profile. Animals monitored by per rectal palpation 10 days apart around day 70-80 postpartum revealed 30 cows to be anestrous or subestrous, which were randomly allocated to five different estrus synchronization protocols at day 90-92, each with six cows and a group of six cows that exhibited estrus and were bred by day 70-90 served as normal control. The treatment protocols included Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and double PGF2α together with estimation of plasma metabolites on day 0, 7, 9/10 (AI) and then at 10 days interval up to day 40 post-AI. 

Result: The mean plasma total protein levels varied non-significantly, while total cholesterol and triglycerides varied significantly (p<0.01) between intervals until 90 days postpartum in most of the groups with gradual increase in the mean values from the day of calving. The estrus synchronization protocols, however. did not alter the plasma profile of these constituents. The estrus induction response with Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α treatment was 66.66, 83.33, 50.00, 66.66 and 66.66% and the conception rates at induced estrus 16.66, 33.33, 16.66, 50.00 and 50.00%, respectively. In normal cyclic control group, the conception rate at first cycle was 33.33%. The plasma total cholesterol and triglycerides were higher in conceived than non-conceived cows, particularly during early postpartum period and post-AI. It was concluded that CIDR, Cosynch and PGF2α protocols were better for estrus induction and conception rate in anestrous and subestrous suckled Kankrej cows, although they did not influence the plasma metabolic profile.

INTRODUCTION

Fertility of the cow in the months following calving depends on satisfactory involution of uterus and re-establishment of cyclical ovarian activity. Prolonged inter-calving period due to delay in onset of postpartum ovarian function is regarded as one of the major reproductive problems responsible for failure to maintain optimum reproductive efficiency, which in turn causes economic loss to the dairy farmers (Shamsuddin et al., 2006; Bhoraniya et al., 2012a; Dhami et al., 2015, 2019; Prajapati et al., 2018). The Kankrej, a dual purpose zebu cattle is known for its endurance, but is slow breeder with very late maturity, prolonged postpartum infertility and thereby calving interval and strong mothering instinct leading to suppression of postpartum ovarian activity. These peculiarities necessitate scientists to initiate appropriate steps to improve their reproductive efficiency from all these angles. Fixed time artificial insemination (FTAI) protocols of estrus induction/ovulation synchronization have been developed to decrease reliance on estrus detection in reproductive management programmes and to improve herd fertility (Pursley et al., 1995; Keskin et al., 2011; Bhoraniya et al., 2012b; Borakhatariya et al., 2017; Dhami et al., 2019).  To establish sustainability in the dairy industry, it is important that cows become pregnant at a biologically optimal time and at an economically profitable interval after calving (Sakaguchi, 2011). The reproductive endocrinology and blood biochemical and mineral profile is also one of the focus to identify specific reproductive and nutritional problems and thereby to adopt appropriate therapeutic measures to augment fertility in dairy animals. The literature on postpartum metabolic profile and fertility without and with hormonal treatments is meagre in zebu cows (Ammu et al., 2012ab; Bhoraniya et al., 2012ab; Naikoo et al., 2016). Hence, the present study was planned to evaluate the postpartum plasma biochemical/metabolic profile till day 90 and then to examine if it can be influenced by different estrus synchronization protocols in postpartum anestrous/subestrous suckled Kankrej cows concurrent with fertility improvement.

MATERIALS AND METHODS

Selection and management of animals
 
The study was conducted during 2011-12 on a total of 36 healthy pluriparous parturient Kankrej cows of the University Livestock Farm, Anand after approval of the IAEC. The cows were monitored from the day of calving till 140 days postpartum. The animals were maintained identically under loose housing system with routine optimum farm feeding schedule and had free access to pure wholesome drinking water. The cows were hand milked twice daily following suckling by the young ones till dry off. Cows were inseminated only after 60 days of calving, if found in estrus. The reproductive/ovarian status of these cows was assessed by palpation per rectum of the genitalia on three occasions, each at 10 days interval beginning at day 70 postpartum, which revealed most of them (n=30) to be in anestrous/ subestrous condition. Jugular blood sampling was done at an interval of 10 days from day of calving till day 90. Based on the reproductive status, the anestrous and subestrous cows were randomly distributed at day 90-92 postpartum into five treatment groups, each with 6 animals and one group (six animals) exhibiting normal estrus within 80 days postpartum served as normal cyclic control. The anestrous and subestrous animals so selected were treated with following controlled breeding techniques.
 
Treatment protocols 90 days postpartum (n=6 each)
 
Group I (Ovsynch protocol)
 
The anestrous cows were injected with 20 μg Buserelin acetate - GnRH analogue (Receptal®, 5 ml) i/m followed by an injection of PGF2α 500 μg (Cyclix®, 2 ml) 7 days later and a second injection of 20 μg GnRH on day 9. Fixed time insemination (FTAI) was performed 22 hrs after second GnRH injection.
 
Group II (CIDR protocol)
 
In anestrous cows, a CIDR (1.38 g progesterone in a nylon spine, Pfizer) was inserted intravaginally and on day 7 the cows were treated with i/m dose of PGF2α 500 μg (Cyclix, 2 ml) accompanied with CIDR removal. FTAI was performed at 22 hrs after PGF2α injection.
 
Group III (Ovsynch plus CIDR protocol)
 
It combined insertion of CIDR intravaginally for 7 days together with administration of Ovsynch protocol in anestrous cows.

Group IV (Cosynch protocol)
 
Here the subestrous cows having established normal cyclicity, but with silent estrus/ovulation till day 90 postpartum were treated with an i/m injection of 20 µg GnRH on day 0 and 500 µg PGF2α on day 7 with FTAI 48 hrs latter simultaneous to second i/m injection of 20 µg GnRH.
 
Group V (PGF2α protocol)
 
The subestrous animals were administered with 500 µg PGF2α i/m twice at 11 days interval and FTAI was performed 72 hrs after the second PGF2α injection.
 
Group VI (Normal cyclic control)
 
Six normal healthy cows exhibiting first pronounced estrus within 3 months postpartum were included in this group.
       
All the cows were bred during spontaneous or induced estrus and were followed for pregnancy or estrus recurrence, if any. The cows returned to estrus were re-inseminated. In non-return cases pregnancy was confirmed per rectum 60 days post-AI. Out of total 36 animals under study, the cows conceived at first AI, irrespective of treatment groups, were classified as conceived or pregnant (n=12) and the remaining cows (n=24), which returned to next estrus, were taken as non-conceived cows.
 
Blood sampling and biochemical estimations
 
Blood samples were collected in heparinized vacutainers through jugular vein puncture from all the animals on the day of calving and thereafter at 10 days interval up to the initiation of synchronization treatment protocols in anestrous/subestrous animals around day 90 postpartum, or AI in case of cyclic control animals. The blood sampling was later rescheduled based on the treatment protocols or AI, i.e., on day of first treatment (day 0), day of PGF2α injection (day 7), day of induced/spontaneous estrus and AI (day 9/11) and then on day 10, 20, 30 and 40 post-AI. The samples were immediately centrifuged at 3000 rpm for 15 minutes and the plasma samples were stored at -20°C by adding a drop of 0.01% sodium merthiolate until analyzed. The concentrations of plasma total protein, total cholesterol and triglycerides were determined using standard procedures and assay kits procured from Crest Biosystem, Goa on biochemistry analyzer (Mindray, M-20).
 
Statistical analysis
 
The estrus induction response and conception rates of different estrus synchronization protocols were compared by Chi-square test (Snedecor and Cochran, 1994). The data on plasma metabolic profile were analyzed using completely randomized design and Duncan’s new multiple range test through an online SAS system of statistical analysis.

RESULTS AND DISCUSSION

Plasma metabolic profile postpartum and in different treatment groups
 
The mean plasma protein concentrations recorded during first 90 days postpartum fluctuated non-significantly between different time intervals in all the groups and between groups at most of the intervals postpartum, except for Ovsynch + CIDR group, where the period effect was significant (Table 1). These observations compared favorably with the report of Ammu et al., (2012a) in Gir cows (5.82±0.19 to 6.44±0.15 g/dl). Similarly, Patel and Dhami (2006) in HF cows and Khasatiya et al., (2007) in Surti buffaloes observed non-significant weekly variation in plasma total protein profile from caving till 21 weeks postpartum without or with single GnRH or PG injection on day 42-49. Onita and Colibar (2009) in contrast recorded increasing trend of total protein from calving till 40 days postpartum. The mean plasma protein concentrations on day 0, day of PG injection and on day of FTAI in various treatment protocols, viz., Ovsynch, CIDR, Ovsynch + CIDR, Cosynch, PGF2α also did not vary significantly, including those of day 12 post-AI, between different intervals or between groups at any of the intervals studied. The values fluctuated non-significantly in a narrow range among groups and periods, including those of control group (Table 1). These findings concurred well with Bhoraniya et al., (2012a) in Ovsynch and CIDR treated anestrous Kankrej cows and Ammu et al., (2012a) in control, CIDR, Ovsynch and Cosynch treated Gir cows during treatment and up to 40 days post-FTAI. Dhami et al., (2007) recorded almost identical values of plasma total protein in anestrous (6.80±0.10 g/dl) and subestrous (6.72±0.09 g/dl) HF cows and the levels were not influenced by the hormone therapy on day 49 postpartum. The weekly profile of plasma total protein also neither differed significantly between days of the cycle nor between fertile and infertile cycles (Kavani et al., 2005), however the values were apparently higher in fertile cycles. Ahmad et al., (2004) and Kumar et al., (2010) reported significantly higher plasma protein levels in cyclic as compared to repeat breeders and/or anestrous cows and buffaloes.
       
The mean total cholesterol concentration differed significantly (p<0.01) between different intervals postpartum in all the groups, The overall mean total cholesterol levels were the lowest on the day of calving, which then increased gradually and significantly till day 40-50 postpartum and then remained more or less static till day 90 and even up to day 140 postpartum (Table 2). The increasing trend of plasma total cholesterol concentration seen from the day of calving to subsequent weeks postpartum, was associated with the initiation of ovarian activity and establishment of estrous cyclicity postpartum in some animals with silent ovulation as evident from rectal palpation and their plasma progesterone profile (Naikoo et al., 2016). Present findings corroborated well with those of Guedon et al., (1999), Dhoble et al., (2004) and Ammu et al., (2012a), who found a linear and significant increase in serum or plasma cholesterol concentration from 2 to 9 weeks after calving in cows, which then declined slightly. Patel and Dhami (2005) also recorded similar observation in lactating and pregnant HF cows. Khasatiya et al., (2005) noted significant weekly variation in serum total cholesterol content of both fertile and infertile groups of Surti buffaloes with gradual increasing trends from calving till 15 weeks postpartum. Onita and Colibar (2009) recorded increased cholesterol concentration attaining a peak value of 320±21.10 mg/dl in 41 days of lactation in cows, more than the physiological level. Further, none of the synchronization treatments significantly influenced the plasma total cholesterol levels and the values were almost constant during various synchronization therapies and thereafter up to 40 days post-AI, although the levels in Ovsynch, CIDR and Cosynch groups were observed to be slightly elevated during and after these therapies. In contrast the levels in control group declined insignificantly and showed an inconsistent pattern over the 40 days post-AI. Ammu et al., (2012a) recorded significant variation in total cholesterol among control, CIDR, Ovsynch and Cosynch treated Gir cows during treatment period and post-AI. Bhoraniya et al., (2012a) also did not find significant variation in the mean values of total cholesterol in anestrous Kankrej cows of Ovsynch, CIDR, Ovsynch + CIDR, Heatsynch protocols on different days or between protocols on any of the days/ periods. Ahmad et al., (2004) didn’t find significant difference in plasma cholesterol concentration between cyclic and non-cyclic cows.
 
The mean plasma triglyceride concentrations during first 90 days postpartum varied between different intervals in all the groups with significant (p<0.05) differences only in Ovsynch, Ovsynch + CIDR, PGF2α and control groups.  For other groups, the period effect was insignificant before, during and after treatment/post-AI till 40 days. The values on the day of calving were the lowest and showed increasing trend till 90 days postpartum (Table 3). Ammu et al., (2012a), Khasatiya et al., (2005) and Kalasariya et al., (2017) found gradual and significant rise in weekly concentration of triglycerides during early postpartum period in Gir cows and Surti buffaloes, respectively. Guedon et al., (1999) opined that the plasma triglycerides levels were influenced by physiological status of animal and were higher during last 10 weeks of pregnancy than at calving and early postpartum. Patel and Dhami (2005) in HF cows noted little lower values of triglyceride at calving, which got slightly elevated from 5th to 6th week postpartum and again declined from 13th-14th week till 21st week postpartum. Further, it was not influenced by GnRH and PGF2α administration around day 49 postpartum. In the present study, except Ovsynch, no other protocol influenced the plasma triglyceride levels and values in all the treated and control groups were almost same before, during and after AI at spontaneous or induced estrus up to day 40 post-AI (Table 3). Ammu et al., (2012a) found insignificant influence of different estrus induction/ synchronization protocols on plasma triglycerides concentration during treatment or till 40 days post-AI in Gir cows. The findings indicate that this trait is not usually influenced by hormonal therapy in dairy bovines.
 
Fertility response to synchronization treatments
 
The estrus induction and ovulation response of cows treatment with  Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α protocols was 66.66, 83.33, 50.00, 66.66 and 66.66%, respectively, as confirmed by presence of CL on the ovary 12 days post-FTAI. Comparable or higher ovulatory response has been reported with one or more of these protocols by many researchers (Mohankrishna et al., 2010; Keskin et al., 2011; Khade et al., 2011; Ammu et al., 2012a; Bhoraniya et al., 2012b; Hadiya et al., 2015; Dhami et al., 2015, 2019; Prajapati et al., 2018) in cyclic and/or acyclic cattle. The conception rates obtained at the induced estrus/FTAI under Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α protocols and in control group were 16.66, 33.33, 16.66, 50.00, 50.00 and 33.33%, respectively. The conception rate at the induced estrus in Ovsynch plus CIDR protocol was at par with Ovsynch alone and was lower than in other groups and even the control group. There was no beneficial effect of combining Ovsynch with CIDR.
 
Mohankrishna et al., (2010) recorded conception rate of 30% in Sahiwal cows, while higher conception rate of 50% each in Gir and Kankrej cows with Ovsynch were recorded by Khade et al., (2011), Ammu et al.. (2012a) and Bhoraniya et al., (2012a). The present conception rate with CIDR and Ovsynch + CIDR were also lower than that recorded by Khade et al., (2011), Bhoraniya et al., (2012b) and Hadiya et al., (2015) in anestrous Gir and Kankrej animals. Patel et al., (2014) recorded comparable conception rate at FTAI following mid-cycle PG injection in repeat breeding crossbred cows, while Chaudhary et al., (2012) found lower conception rates at FTAI with doublesynch and estradoublesynch protocols in post-pubertal anestrus Gir heifers. The prolonged exogenous progesterone priming from CIDR device caused negative feedback effect on hypothalamo-hypophyseal-gonadal axis and increased receptors for gonadotropins on the ovaries followed by rebound on its sudden withdrawal causing stimulated FSH secretion, folliculogenesis and ovulation. The present findings suggest that the CIDR insert alone over the Ovsynch or its combination improved synchrony of estrus occurrence and considerably enhanced conception rate in anestrous suckler Kankrej cows. Further, the estrus response following Cosynch and double PG protocols in subestrous Kankrekj cows was identical, but the conception rate achieved at FTAI was better with Cosynch (50.00 vs 33.33%). Almost similar were the observations of Orkun et al., (2006), Patel et al., (2014), Hadiya et al., (2015) and Prajapati et al., (2018) in exotic and crossbred cows.
 
Metabolic profile of conceived and non-conceived cows
 
The mean plasma total protein content neither of conceived (n=12) nor of non-conceived (n=24) groups differed significantly between 10 days intervals postpartum. The values were lowest on the day of calving in both conceived (6.00±0.13 g/dl) and non-conceived (5.85±0.14 g/dl) groups and then showed slight elevated levels from day 10 to 140 postpartum with maximum values of 6.70±0.18 and 6.62±0.10 g/dl on day 60 postpartum in conceived and non-conceived group (Fig 1). The non-significant difference could be due to optimal and uniform nutritional status of all animals under farm conditions. The present findings on conceived and non-conceived groups were in favour of report of Srivastava and Sahni (2000) who did not find any correlation of total protein with fertility status in cows and buffaloes, though the values were higher in conceived than non-conceived group.  Patel and Dhami (2006) also couldn’t find any significant difference in the level of protein between fertile and infertile cycles or between conceived and non-conceived HF cows and the values fluctuated non-significantly over 21 weeks postpartum period, which matched with the present findings. Dhoble et al., (2004), however, recorded significantly higher level of plasma protein in pregnant than non-pregnant animals (7.92 vs 7.71 g/dl).

There was no significant difference in the values of plasma total cholesterol between conceived and non-conceived groups at any of the intervals up to 90 days postpartum. Thereafter, there was insignificant rise in the values of conceived group compared to that of non-conceived group (Fig 1). Cholesterol being immediate precursor of steroid hormones, early rise around day 30-40 postpartum concomitant to rise in plasma progesterone in conceived cows in association with follicular activity, though with silent ovulations, is convincing (Naikoo et al., 2016; Dhami et al., 2017). The present findings corroborated well with the observations of Patel and Dhami (2005) in HF cows for conceived and non-conceived groups over 21 weeks postpartum with highly significant (p<0.01) differences between weeks from 62.40±9.19 to 115.27±6.79 mg/dl. Dhoble et al., (2004) recorded significantly higher level of cholesterol in pregnant than non-pregnant cows, while Ammu et al., (2012b) reported that the total cholesterol levels were not influenced by conceived and non-conceived status in Gir cows. Patel et al., (2005), however, recorded significantly lower mean cholesterol level in conceived than non-conceived groups of repeat breeding cows, though the weekly values didn’t vary between 7 weeks period post-treatment. Sivaraman et al., (2002) did not find significant difference in serum total cholesterol levels of lactating and lactating pregnant Jersey crossbred cows, but it was significantly lower in dry non-pregnant cows. Kumar et al., (2009) reported serum total cholesterol in normal cyclic and repeat breeding cows as 279.94±26.33 and 215.31±10.66 mg/dl, respectively, without significant variation between conceived and non-conceived groups.
 
There was significant variation in plasma triglycerides between conceived and non-conceived cows up to 90 days postpartum, although the profile showed slight rise from calving with advancing postpartum period in both the groups (Fig 1). Dhoble et al., (2004), Patel et al., (2005) and Ahlawat and Derashri (2009) recorded higher levels of triglycerides in pregnant/ conceived than non-pregnant/non-conceived cows. However, Khasatiya et al., (2005), Patel and Dhami (2005) and Dhami et al., (2017) in Surti buffaloes, HF cows and crossbred cows, respectively, did not find any significant difference in the values of triglycerides between conceived and non-conceived groups. Dhami et al., (2005) recorded an overall mean plasma triglycerides value of 19.17±1.15 mg/dl among 72 HF animals, which differed significantly in decreasing order between advanced pregnant (19.46±3.41), recently calved, lactating, anestrous and repeat breeding (9.56±2.16 mg%) groups.

CONCLUSION

The study on plasma metabolic profile particularly total cholesterol and triglycerides during postpartum period could assess the postpartum nutritional status and ovarian activity in suckled Kankrej cows. Estrus induction and improvement in conception rate is possible in postpartum anestrous/suestrous suckled cows with the use of different synchronization protocols. CIDR protocol resulted in better estrus expression and fertility at FTAI in anestrous cows and Cosynch protocol in subestrous cows as compared to other protocols in infertile Kankrej cows. The linear increasing trend of plasma total cholesterol  observed over the postpartum period with higher values in fertile/ conceived than infertile/ non-conceived animals might be attributed to early resumption of follicular activity with steroidogenesis and thereby conception in former group.

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