SOME HORMONAL AND BIOCHEMICAL BLOOD INDICES IN PREGNANT BUFFALOES BEFORE AND AFTER RETENTION OF PLACENTA

SOME HORMONAL AND BIOCHEMICAL BLOOD INDICES IN PREGNANT BUFFALOES BEFORE AND AFTER RETENTION OF PLACENTA

AMIRA M.M.METWALLY *; ENAS A.H. FARAG* and MAAROUF A.A.**

* Dept. of Biochemistry, Animal Health Research Institute, Benha branch.

** Dept of Microbiology, Animal Health Research Institute, Benha branch

INTRODUCTION

Retained placenta is the main common reproductive disorders in dairy buffaloes which induced subsequent uterine infection, endometritis, reduction of milk yield and lower conception rate beside decrease market value of animal (Hemingway, 2003 and Semacan and Sevinc, 2005). Although the etiology of retained placenta has been the subject of numerous studies the exact cause is not clear (Joosten and Hesen, 1992 and Wischral et al., 2001). Meanwhile, it was found that various causes of RP have been identified i.e uterine paresis, abortion, late or pre-mature birth, dystocia infections and hormonal disorders beside some vitamin and minearal deficiencies predispose for RP (Ahmed et al., 2009 and Akar and Yeldiz, 2005). According to Brzezinska-Slebodzinsk et al. (1994) and Kankofer (2001 a.) oxidative stress in cow is a contributory factor to increase disease susceptibility since metabolic demand associated with late pregnancy and increase milk production would be expected to increase production and uncontrolled increase reactive oxygen species (ROS) which could be a risk factor for RP (Sordillo, 2005). RP was reported to be associated with oxidative stress in dairy cows (Kankofer, 2001 b). The antioxidantvitamins (e.g vitamin E) is important for improve the fertility of dairy cows(Allison and Laven, 2000). In late pregnancy , the production of free radicals wasincreased, these elevation in free radical and increasing in oxidativedamage may involved in parturition, meanwhile the over increase in the oxidativeprocess after parturition may led to RP and /or postpartum infertility (Toescu et al., 2002). However vascular changes and uterine contractions has been found to play a role in placental release (Laven and Peters, 1996).

This study is a trial to highlight on some parameters as indicators of RP in dairy buffoles.

MATERIALS and METHODS

Animals and their managements: Thirty two pregnant dairy buffaloes belonging to private dairy farm in Kaliobeia Governorate were used in this study. Animals age 4-7 years and were kept under veterinary supervision, general health condition were examined, housed in the same feeding and management condition until end of study. Case history of each animal was recorded. Buffaloes were considered to retain their placenta if it does not drop after at least 12 hrs post calving (Stephen, 2008).Buffaloes with RPshowed discolored vulval membranes, foul smelling discharge, in appetence and decrease milk yield.

Sampling: Two blood samples (with and without EDTA) were collected from each animal via jugular vein puncture two week, week before parturition and one week after parturition. Uncoagulated blood samples were used for performing complete blood picture as well as for determination of selenium (Se) values. Serum was separated from coagulated blood samples by centrifugation for assaying biochemical constituents.

Analysis: Complete blood picture including erythrogram and leucogram was carried out according to (Jain, 2000).The serum levels of cortisol, progesterone and oestradiol-17β were determined as described by (Hasler et al., 1976; Xing et al., 1983 and Kubosik, 1984) respectively using validated radioimmunoassay (Diagnostic Products Corportation, Los Angles CA). Trace elements including Zn,Fe and Cu concentrations in diluted serum samples and Se in whole blood samples were determined using atomic absorption spectrophotometry  as outline by Varley et al. (1980). Serum calcium was determined according to Gindler and King (1972). Oxidant and antioxidant markers including LPO were estimated according toBuege and Aust (1977), NO (Montgomery and Dymock 1961), CAT (Aebi, 1984), ASCA (Haris and Ray, 1945), R-GSH (Beutler et al., 1963) and TAC (Koracevic et al., 2001) were determined.

Statistical analysis: Data were expressed as means ± standard error (M±SE). The comparison between the groups was conducted by one way analysis of variance (SPSS, 1993).

RESULTS

The results of clinical examination of thirty two pregnant buffaloes (two week, week preparturient and week postparturient) showed that seven out of these animals suffered from RP, two of them had history of retained fetal membrane. which retained their placentae for more than 12 hrs post calving and were considered to suffer from RP and the remaining animals dropped their placentae normally within 12 hrs most of the retained groups were subjected to manual removal of their placentae. All data concerning heamatological results were presented in Table (1) while other markers of oxidant, antioxidant and some biochemical in RP showed Table (2).

Table 1: Mean values of some heamatological parameters of pregnant buffaloes before and after retained fetal membrane.

Values (a-b-c-d) at the same raw followed by different superscript letters were significantly different (P< 0.05).

Table 2: Mean values of some hormonal and biochemical parameters of pregnant buffaloes before and after retained fetal membrane.

    Values(a-b-c-d) at the same raw followed by different superscript letters were significantly different (P< 0.05)

DISCUSSION

Retained placenta is identified as major importance reproductive abnormality of cows and buffaloes. It can be substantial risk factor for toxic puerperal metritis, it couses significant economic losses, as many RP buffaloes develop metritis and suffer from infertility (Azawi and Taha 2002 and Azawi 2006).In this study animals with RP suffered from anemia as indicated by the significant decreasein RBCs, Hband PCV, also there is a leucocytosis accampanied by lymphopenia, neutrophilia and monocytosis these may be attributed to inflammation and increase monocytes for scaveningof celldebris(Sivaraman    et al., 2003). Increase progesteron in preparturient period in our results (Table 2) could be a major factor contributing to the pathogenesis  of retained fetal membrane (RFM) in buffaloes, however normally increasing estrogen and decreasing progesterone activities lead to multiple changes which is accompanied by increasing  prostaglandin synthesis and mainly toarise oxytocin receptor concentration in the myometrium these basic changes are necessary for subsequent placental separation and expulsion (Husslein, 1984; Sabry et al., 1997 and Michal and Hanna, 2006). Increased progesterone level in RP may be due to failure of placenta to produce specific steroidal enzymes that help in progesterone metabolism and its conversion to esterogen (Matton and Dufour, 1987), which in turn may induce the accumulation of immunosuppressive proteins in the uterine lumen and make the uterus susceptible to infection and persistence bacteria (Konigsson et al., 2002).

It has been documented that RFM associated with the presence of uncontrolled eleveted level of reactive oxygen species (ROS) which may disturb physiological processes leading to expulsion of the placenta (Kankofer, 2001 a,b), when free radical generation exceeds the body^’s antioxidant production capacity, oxidative stress develops Roth, (2000) reported that estradiol has a potant antioxidant activities. The periparturient significant increase in estradiol-17β concentration is in agreement with that results of Eissa et al. (1995) and Badr et al. (2001).

The rise in cortisol (Table 2) stimulates the conversion of placentally derived progesterone to estradiol by activating the placental enzyme17α - hydroxylase which hydroxylates progesterone via androstenedione to estradiol 17β. The consequences of the rise in estradiol17β and decline of progesterone in prepheral circulation have a triple effects, first estradiol has a direct effect upon the myometrium increasing its responsiveness to oxytocin, second they produce softening of cervix by altering the structure of collagen fibers, third they act upon the cotyledon-caruncle complex to stimulate the production and release of PGF_2α (Grunet, 1986 and Horta, 1988).

In the current study it was found that peroxidative indices (LPO and NO) are an indicator of over production of free radicals and cellular damage which were elevated during the last two weeks of pregnancy compared with non pregnant animals in the same time the level of antioxidant (CAT, ASCA, SOD TAC, selenium and vitamin E) were decreased these mean that oxidative stress and exhaustion of the antioxidant were involved in the late stage of pregnancy. These resultsagree withMegahed et al. (2006); Ahmed      et al. (2009) and Toescu et al. (2002)who reportedthat late pregnancy was associated with the formation of susceptible oxidisable particles and an increase in oxidative damage. Antioxidative defense mechanisms are complex net work used to scavenge ROS. Kankofera et al. (2005) recorded that RP in buffaloesis related toimbalance between production andneutralization in ROS. Moreover, it was suggested that alterations inantioxidative capacity are related to changes in hormonal levels which appear close to parturition. Clinically important symptoms are defined as the disturbances in steroid hormones as well as prostaglandin F_{2 α} concentration (Leidl and Rockel, 1980 and Heuwieser and Grunert, 1987). In this study  significant decrease was recorded (Table 2) in selenium,vitamin E, Zn, Cu and Fe, similar finding were reported by Segerson et al. (1980); Ahmed et al. (1999) and EL-Khadrawy and EL-Ekhnawy (2005 )who found that inadequate concentrations of selenium and vitamin E in the diet increased oxidative stress, production of lipid peroxidase and the incidence of RFM. Se, Cu and Zn are trace minerals required for functioning of enzymes involved in the antioxidant defense system. Se is known to be incorporated in the glutathione peroxidase performing the antioxidativedefense of the body by eliminating hydrogen peroxides(Kommisrud and Vatn, 2005). Cu and Zn are involved in the antioxidant system via its involvement in superoxidase dismutase (SOD) and ceruloplasmin. Copper, Zn and SOD are responsible for dismutation of superoxide radicals to hydrogen peroxidein the cytosol(Halliwell and Gutteridge, 1999).

It could be concluded that retention of placenta is a serious problem resulted in uterine infection and toxic puerperal metritis. Also it has an adverse effect on some hormonal and biochemical parameters. So supplementation of dam with a balanced ration containing adequat amount of antioxidant during late stage of pregnancy beside that sufficient daily exercise, sufficient large clean and comfortable calving areas and proper sanitary procedures during the calving period is advised to minimize PR and infection of the reproductive tract.

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