Expression and Localization of Vascular Endothelial Growth Factor and its Receptors in the Corpus Luteum During Oestrous Cycle in Water Buffaloes (Bubalus bubalis)

The aim of this study was to document the expression and localization of VEGF system comprising of VEGF isoforms (VEGF 120, VEGF 164 and VEGF 188) and their receptors (VEGFR1 and VEGFR2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors. In general, all the components of VEGF system (the VEGF isoforms and their receptors) were found in the water buffalo CL during the oestrous cycle. The mRNA as well as protein expression of VEGF system was highest during the early and mid‐luteal phase, which later steadily decreased (p < 0.05) after day 10 to reach the lowest level in regressed CL. As demonstrated by immunohistochemistry, VEGF protein was localized predominantly in luteal cells; however, VEGFR1 and VEGFR2 were localized in luteal cells as well as in endothelial cells. In conclusion, the dynamics of expression and localization of VEGF system in buffalo corpora lutea during the luteal phase were demonstrated in this study, indicating the possible role of VEGF system in the regulation of luteal angiogenesis and proliferation of luteal as well as endothelial cells through their non‐angiogenic function.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

Immunoexpression of cytokine tumour necrosis factor-α suggesting its role in formation and regression of corpus luteum in Indian buffalo

Tumour necrosis factor-α (TNF-α) is a cytokine that plays multiple important roles in corpus luteum (CL). Immunolocalization of expression of TNF-α in CL of buffalo was studied in different stages of its development and regression. Corpus luteum of healthy buffaloes (24) was collected from local slaughterhouses and categorized into early (stage I, 1–5 days, n = 6), mid (stage II, 6–11 days, n = 6), late luteal (stage III, 12–16 days, n = 6) and regressing phase (stage IV, 17–20 days, n = 6). In earliest phase of cyclic CL, per cent immunoexpression of TNF-α was significantly (p < .05) lower as compared to all phases with its expression being restricted to few developing luteal cells, usually in neutrophils. A significantly (p < .05) higher number of neutrophils with TNF-α immunoexpression were observed as compared to mid-luteal phase that indicated its role in initiation of angiogenesis at this stage. TNF-α immunoexpression almost doubled in mid-luteal phase, but the number of neutrophils exhibiting TNF-α was significantly (p < .05) lower with respect to all phases of CL. Immunoexpression percentage in late luteal phase increased sharply being significantly (p < .05) higher than earlier two phases of CL. In regressing phase, per cent immunostaining was maximum with highly significant (p < .05) difference as compared to all other stages, observed in all degrading luteal cells, abundant immune cells, that is neutrophils and macrophages which finally led to apoptosis and phagocytosis. Immunoexpression of TNF-α in early luteal phases served its role in initiation of angiogenesis, and its intense expression in regressing phase of CL suggested a shift in its role to apoptosis and structural luteal regression signifying both luteotropic and luteolytic roles in buffalo. This is probably the first study of its kind in buffaloes.     

Endometrial glycogen, protein, nucleic acids and phosphatases during oestrous cycle in buffaloes (Bubalus bubalis).

Distinct cyclic variations were observed in the concentrations of endometrial glycogen, protein, deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) and activities of acid phosphatase (ACP) and alkaline phosphatase (AKP) during the follicular and luteal phases of the oestrous cycle in buffaloes. Glycogen, protein, DNA and RNA concentrations were significantly (P less than 0.01) higher in buffalo endometrium during the follicular phase than the luteal phase of the oestrous cycle whereas ACP and AKP activities were significantly (P less than 0.01) elevated during the luteal phase compared with the follicular phase. These results are discussed in relation to the nourishment of the blastocyst during the pre-implantation period.     

A study on the ovarian follicular dynamic in Iraqi Northern Buffaloes

The aim of the present study was to test the hypothesis of wave pattern of follicular growth and to monitor the ovarian follicular dynamic in Iraqi buffalo cows. Reproductive tracts were collected at random intervals slaughtered at Mosul abattoir. According to morphological appearance of the corpus luteum, the estrous cycle was divided into four stages. The number of subordinate follicle (<5 mm in diameter) was higher during stage 1 (metestrous) and stage (proestrous and estrous) than during other stages of the estrous cycles, 13.5 ± 6.08 and 4 9.41 ± 3.94, respectively. There were fewer follicles (5–8 mm in diameter) during early diestrous and proestrous, 1.66 ± 1.42 and estrous, 0.69 ± 0.47 than during metestrous, 4.53 ± 3.23 and late diestrous, 3.66 ± 2.23. Follicles > 8–12 mm in diameter were more numerous during early diestrous, 1.62 ± 1.29 and late diestrous, 1.03 ± 0.72. A total 38 (64.6%; 82/127) animals examined showed follicles larger than 8 mm during early and late diestrous (stage 2 and stage 3). This indicated that these buffaloes developed two follicular waves in their cycle. Buffaloes did not show follicles larger than 8 mm during early and late diestrous were 45 animals (35.4%; 45/127), but all of these presented one large follicle during the following stage. These buffaloes develop only one follicular wave in their cycle. It could be concluded that, 64.6% of Iraqi buffalo cows develop two patterns of follicular waves, and 35.4% showed one wave of follicular dynamics.     

Effect of ruminally protected Methionine on the productive and reproductive performance of grazing <Emphasis Type="Italic">Bos indicus</Emphasis> heifers raised in the humid tropics of Costa Rica

With the objective of evaluating the effect of methionine supplementation prior to a breeding program, thirty one heifers (Bos taurus x Bos indicus) were used averaging 386  ±  29 days of age and a mean body weight of 402.6  ±  28 kg. Fifteen of the animals received a supplement (SG) during 45 days with molasses-urea mixture (2 kg molasses + 407 g urea/head/day), plus 10 g of ruminally protected methionine. The other sixteen heifers did not receive supplement (CG). Fecal and pastures samples were collected to assess dry herbage intake and digestibility. Serial ultrasound measurements from the ovary were performed in both groups to evaluate follicular dynamics. The heifers were categorized according to their follicular size and presence of a CL. Forage intake and dry matter digestibility were reduced (P  <  0.05) and body condition tended to improve (P  =  0.07) in the supplemented heifers, however, total intake, final weight, daily gain and dorsal back fat were not affected. After the supplementation period, the percentage of females in the categories < 3 mm and 3 to < 6 mm, was greater (P  <  0.05) in CG (25% and 43.7%) that in SG (0% and 26.6%) but in the follicle category of ≥  9 mm, the percentage of animals was 60% in SG and 18.8% in CG (P  <  0.05). The percentage of ovulation for the SG and CG was 86.7% and 62.5%, respectively (P  <  0.05). The combination of supplementation with methionine-urea and molasses at the end of the dry season and the anticipated onset of the rainy season favored the establishment of ovarian activity and follicular dynamics.     

Comparative therapeutic effect of moxidectin,doramectin and ivermectin on psoroptes mites infestation in buffalo (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>)

The aim of the present study was to carry out comparative therapeutic effect of moxidectin pour on, doramectin and ivermectin on psoroptes infestation in buffalo. A total of 318 buffalo in 77 small scale herds suspected to have mange mites were examined clinically and parasitologically. Fifty-three (16.66%) buffalo in 25 herds were recorded to be infested; 51 (16.35%) with psoroptic mites, and two (0.31%) with chorioptic mites. Buffalo with psoroptic mites were randomly allocated into three groups (17 buffalo each). First group was treated with moxidectin pour on at a dose rate of 0.5 mg kg^-1. The second group received doramectin (200 μg kg^-1 twice subcutaneously, 14 days apart). The third group received ivermectin (200 μg kg^-1 twice subcutaneously, 14 days apart). Adjunct to each drug, deltamethrin was applied to the surrounding environment twice at a two week interval. Treatment outcomes of 51 buffalo with psoroptic mites showed that moxidectin pour on and doramectin had a significant higher effect on mite count reduction (MANOVA, P < 0.01; Walks^, Lambda, P < 0.01) and clinical sum scores (MANOVA, P < 0.05; Walks^, Lambda, P < 0.05) compared with injectible ivermectin. On clinical level, the number of clinically recovered buffalo in moxidectin and doramectin treated groups was significantly (P < 0.05) higher than that of ivermectin treated group. The result of the present study indicated that psoroptic mites are the main cause of mange in buffalo in Lower Egypt. This is the first report that describes the effect of moxidectin in buffalo. Moxidectin is a good alternative and easily applied drug for treatment of psoroptes infestation in buffalo.     

Follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in water buffalo (Bubalus bubalis)

The objective of this study was to examine the follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in buffaloes. Ovaries were collected from 56 acyclic and 95 cyclic buffaloes at slaughter, surface follicle number was counted and follicles were classified into small (5.0–6.9 mm), medium (7.0–9.9 mm), and large (≥10.0 mm) size categories based on their diameter. Follicular fluid was aspirated and assayed for nitric oxide, ascorbic acid, estradiol, and progesterone. Acyclic buffaloes had a higher (P < 0.05) number of medium-sized follicles and a lower (P < 0.001) number of large follicles than the cyclic ones. In acyclic animals, the number of large follicles was lower (P < 0.01) than in medium size category which in turn was lower (P < 0.001) than the number of small follicles. In contrast, the number of medium and large follicles was not different (P > 0.05) in the cyclic control. However, the number of small-sized follicles was higher (P < 0.001) compared to the other two categories. The incidence of large-sized follicles was lower (P < 0.05) in acyclic buffalo population compared to the cyclic control. Evaluation of estrogenic status demonstrated that all the follicles of acyclic buffaloes are estrogen-inactive (E ₂/P ₄ ratio < 1). Small- and medium-sized follicles of acyclic buffaloes had higher concentrations of nitric oxide (P < 0.05 and P < 0.001, respectively) and lower concentrations of ascorbic acid (P < 0.05 and P < 0.01, respectively) than the corresponding size estrogen-active follicles of their cyclic counterparts. In conclusion, this study indicates that follicular development continues during acyclicity in buffaloes. Although follicles in some acyclic buffaloes attain a size corresponding to morphological dominance, they are unable to achieve functional dominance, perhaps due to an altered balance of intrafollicular nitric oxide and ascorbic acid and, as a result, these follicles instead of progressing to ovulation undergo atresia.     

Relationship between endometritis and oxidative stress in the follicular fluid and luteal function in the buffalo

In this study, alteration in the follicular fluid composition and luteal function was investigated in the buffalo with endometritis. Genitalia were classified into cytological and purulent endometritis on the basis of polymorphonuclear cell cut off while non‐endometritis served as control (n = 10/group). In the follicular phase, the number of surface follicles was counted, diameter of the largest follicle was measured and the follicular fluid was assayed for total protein, cholesterol, malondialdehyde (MDA), total antioxidant capacity (TAC), oestradiol (E₂) and progesterone (P₄). The P₄ content of corpus luteum during mid‐luteal phase was estimated by radioimmunoassay. Ovaries from the follicular phase of oestrous cycle showed no significant difference in the total number of surface follicles, size of the largest follicle and volume of follicular fluid in the buffaloes with and without endometritis (p > .05). However, the antral fluid of the largest follicle from the genitalia of buffalo with cytological and purulent endometritis showed a significant decrease in the concentration of total protein, cholesterol, TAC and E₂ and a significant increase in the concentration of MDA and P₄ (p < .05). The results indicated that there is an association between endometritis and decreased ovarian function.     

Immunolocalization of INSL3 in dog foetal Leydig cells and the LGR8 receptor in the gubernaculum testis

Oxidative stress parameters; thiobarbituric acid reaction substances (RBC-TBARS), catalase (RBC-CAT) and reduced glutathione (RBC-GSH)) and the intraerythrocytic concentrations of electrolytes; sodium and potassium (RBC-Na and RBC-K) were determined in 18 well- controlled (WC) and 22 poorly-controlled diabetic mellitus (DM). Dogs with DM had significant higher blood glucose concentration (P < 0.001), haemoglobin A1c (P < 0.01) and fructosamine (P < 0.001) compared to normal healthy dogs (n = 19). Diabetic dogs in both groups had higher RBC-CAT (P < 0.05) while RBC-TBARS were higher significantly only in poorly-controlled DM group (P < 0.05). The RBC-K was significantly higher in both DM groups (P < 0.001). No changes in RBC-GSH and RBC-Na were found between DM and control healthy dogs. By linear regression analysis, the relationship were found between degree of diabetic mellitus and RBC-CAT, RBC-TBARS, RBC-Na and RBC-K. The relationship was also found between oxidative stress parameters and intraerythrocytic K⁺. The results suggest that in diabetic dogs, oxidative stress occurs which related to the severity of disease and may affect potassium homeostasis.     

Changes in Ovarian Follistatin Levels During the Oestrous Cycle in Sheep may Serve as an Intraovarian Regulator

The expression and concentration of follistatin and activin change during oestrous cycle suggesting their involvement in the regulation of follicular development. The aim of this study was to determine the level, source and potential role of follistatin in the sheep ovary. Follistatin in ovarian venous blood, measured by radioimmunoassay, remained at its low level from follicular phase (day ?1 and 0) to mid‐luteal phase (days 11–13) phase but were significantly elevated during the late luteal phase (days 14 and 15) when corpora lutea underwent regression. Western blot analyses of follicular fluid at day 15 of the cycle showed two strong bands at 42 and 45 kDa and weakly stained bands at 39 and 31 kDa. At day 0, these bands became weaker and the 39 kDa band became undetectable. However, there were no differences in follistatin concentrations between ovaries with and without functional corpus luteum (CL) during the whole luteal phase. In addition, although the ovaries of Booroola ewes normally contain more corpora lutea than those of normal merino ewes, follistatin concentrations in both jugular and ovarian venous blood were similar in Booroola and normal merino ewes. It is concluded that the secretion of follistatin from the ovary is not related to the formation of CL or high ovulation rate of Booroola ewes. The elevation in follistatin concentration in follicular fluid and ovarian blood during late luteal phase may indicate a dual role of follistatin in the luteolysis of existing CL and development of new follicle cohort.     

Ion,Protein, Phospholipid and Energy Substrate Content of Oviduct Fluid During the Oestrous Cycle of Buffalo (Bubalus bubalis)

The aim of this research was to analyse the composition of oviduct fluid (ODF) in buffalo cows at different oestrous cycle phases to fulfil the requirements of buffalo embryos in vitro. ODF was collected by chronic cannulation from three cows that were synchronized by administering a synthetic prostaglandin. Based on hormonal profiles, the pre‐ovulatory, ovulatory, post‐ovulatory and luteal phases of the oestrous cycle were defined. The volume of ODF produced (ml/24 h) was influenced by the oestrous cycle, with values (mean ± SE) around ovulation (1.0 ± 0.2) greater (p < 0.05) than in both the luteal (0.4 ± 0.1) and the post‐ovulatory phases (0.5 ± 0.1), but not different from the intermediate values in the pre‐ovulatory phase (0.8 ± 0.2). Among cycle phases, no differences were found in sodium, potassium, calcium and magnesium concentrations (130.0 ± 1.1, 5.1 ± 0.3, 2.8 ± 0.1 and 0.59 ± 0.04 mmol/l respectively). Interestingly, the chloride secretion (μm /24 h) was higher (p < 0.05) at ovulation (150.2 ± 16.5) than during both the luteal (73.7 ± 22.0) and the post‐ovulatory phases (63.7 ± 11.2), with intermediate values in the pre‐ovulatory phase (113.4 ± 23.5). Glucose concentration (mmol/l) was higher (p = 0.056) in the pre‐ovulatory phase (0.06 ± 0.02) than in the luteal (0.02 ± 0.01) and post‐ovulatory (0.02 ± 0.01) phases but not different from values in the ovulatory phase (0.04 ± 0.02). Concentrations of pyruvate and lactate among oestrous cycle phases were similar (0.08 ± 0.01 and 1.0 ± 0.1 mmol/l respectively). The total quantity of phospholipids (μmol/24 h) was greater (p < 0.05) at ovulation (0.21 ± 0.02) compared with the luteal, pre‐ovulatory and post‐ovulatory phases of the cycle (0.09 ± 0.02, 0.13 ± 0.02 and 0.09 ± 0.01 respectively). No differences were found in either the protein concentration (1.8 ± 0.3 mg/ml) or the quantity of proteins secreted in 24 h (1.8 ± 0.4 mg) among oestrous cycle phases. In conclusion, this study provides the first characterization of buffalo ODF during the oestrous cycle, showing species‐specific differences that may be useful for developing suitable media for buffalo in vitro embryo production.     

Vitamin A and β-Carotene Levels in Plasma, Corpus Luteum and Follicular Fluid of Cyclic and Pregnant Cattle

This study was carried out to examine the relationship between the corpus luteum (CL) weight, CL and follicle diameters and progesterone, β‐carotene and vitamin A levels in reproductive organs of cattle obtained from the slaughterhouse. The β‐carotene and vitamin A levels were determined in plasma, CL and follicular fluid (FF) using a spectrophotometric method at different stages of the oestrous cycle (n=40) and at 3–6 months of pregnancy (n=10). The diameters of the CL and follicle were measured using ultrasonography. Plasma progesterone concentrations were determined by an enzyme immunoassay method. The vitamin A levels of the plasma, CL and FF were not related to each other. The highest plasma vitamin A levels were observed in the proestrus and oestrus, at which periods follicular activity dominates. The vitamin A levels in the CL and FF were negatively related to the weight and diameter of the CL and the diameter of follicle, respectively. In contrast to vitamin A, β‐carotene concentrations of plasma, CL and FF were significantly correlated with each other. The highest β‐carotene levels in the plasma, CL and FF were found during pregnancy when there is maximal luteal function, and the β‐carotene level of the CL was significantly correlated with the weight and diameter of CL. Furthermore, the intrafollicular β‐carotene level was negatively correlated with the follicle diameter. There was a positive correlation between plasma progesterone level and the weight and diameter of the CL, but a negative correlation between plasma progesterone level and follicle diameter. Moreover, plasma, FF and CL β‐carotene levels were positively correlated with plasma progesterone levels. This study revealed that β‐carotene levels in the plasma, CL and FF were influenced by the stage of the oestrous cycle or the pregnancy and were related to bovine luteal function without depending on vitamin A.     

Chemical composition of Mopane worm sampled at three sites in Botswana and subjected to different processing

Effects of site and degutting on chemical composition of mopane worm were investigated. Samples were cooked for 30 min in either brine or in plain water. Samples from Moreomabele were high in acid detergent fibre (ADF) (P < 0.05) and acid detergent insoluble nitrogen (ADIN) (P < 0.01) while samples from Sefophe had least ADF and ADIN concentrations. In vitro true dry matter digestibility (IVTDMD) tended (P = 0.06) to be high in samples from Maunatlala than those from Moreomabele and Sefophe. No site difference (P > 0.05) was observed on the rest of the variables. Degutted samples had high crude protein (CP), ADF and IVTDMD (P < 0.05) but lower (P < 0.001) concentrations of ash, acid detergent lignin (ADL) and condensed tannins. Acid detergent insoluble nitrogen was significantly higher (P < 0.001) in degutted than whole samples. Salting did not (P > 0.05) change concentrations of all but ADF which tended (P = 0.09) to be high in salted samples. Samples which were degutted and salted had higher (P < 0.01) ADF and ADIN than degutted and salted or those left whole and salted or left unsalted. Leaving samples as whole diluted the concentration of CP but increased the fibre components and condensed tannins content. However, mopane worm destined for livestock feeding may be left whole and salt added.     

Expression of Lymphangiogenic Vascular Endothelial Growth Factor Family Members in Bovine Corpus Luteum

The aim of this study was to evaluate mRNA expression, protein concentration and localization of the assumedly important lymphangiogenic factors VEGFC and VEGFD and the receptor FLT4 in bovine corpora lutea (CL) during different physiological stages. In experiment 1, CL were collected in a slaughterhouse and stages (days 1–2, 3–4, 5–7, 8–12, 13–16, >18) of oestrous cycle and month <3, 3–5, 6–7 and >8 of pregnancy. In experiment 2, prostaglandin F2α (PGF)‐induced luteolysis was performed in 30 cows, which were injected with PGF analogue on day 8–12 (mid‐luteal phase), and CL were collected before and 0.5, 2, 4, 12, 24, 48 and 64 h after PGF injection. The mRNA expression was characterized by RT‐qPCR. All three factors were clearly expressed and showed significant changes during different groups and periods examined in both experiments. Protein concentrations of VEGFD and FLT4 measured by ELISA were not detectable in early cyclic CL but increased to higher plateau levels during pregnancy. After PGF‐induced luteolysis FLT4 protein showed an increase within 2–24 h after the injection. FLT4 localization by immunohistochemistry in the cytoplasm of luteal cells was relatively weak in early CL. It increased in late CL and especially in CL during pregnancy. During pregnancy, a positive FLT4 staining in both the nucleus and cytoplasm of lymphatic endothelial cells in peripheral tissue was observed. In conclusion, our results lead to the assumption that lymphangiogenic factors are produced and regulated in CL and may be involved in mechanisms regulating CL function, especially during pregnancy.     

Immunohistochemical Localization of Luteinizing Hormone Receptor in the Cyclic Gilt Ovary

Luteinizing hormone receptor (LHR) is a specific membrane receptor on the granulosa and theca cells that bind to luteinizing hormone (LH), resulting in androgen and progesterone production. Hence, the regulation of LHR expression is necessary for follicle maturation, ovulation and corpus luteum formation. We examined the immunolocalization of LHR in cyclic gilt ovaries. The ovaries were obtained from 21 gilts aged 326.0 ± 38.7 days and weighing 154.6 ± 15.7 kg. The ovarian tissues were incubated with rabbit anti‐LHR polyclonal antibody. The follicles were categorized as primordial, primary, preantral and antral follicles. Ovarian phase was categorized as either follicular or luteal phases. The immunolocalization of LHR was clearly expressed in primary, preantral and antral follicles. LHR immunostaining was detected in the cytoplasm of granulosa, theca interna and luteal cells. LHR immunostaining was evaluated using imaging software. LHR immunostaining in the theca interna cells in antral follicles was almost twice as intense as that in preantral follicles (65.4% versus 38.3%, P < 0.01). LHR immunostaining was higher in the follicular phase than in the luteal phase (58.6% versus 45.2%, P < 0.05). In conclusion, the expression of LHR in the theca interna cells of antral follicles in the follicular phase was higher than in the luteal phase. The expression of LHR in all types of the follicles indicates that LHR may impact follicular development from the primary follicle stage onwards.     

Changes in colostrum of Murrah buffaloes after calving

Colostrum samples were collected from 8 Murrah buffaloes on days 1, 2, 3, 4 and 5 after calving. Levels of IgG averaged 54.0 mg/ml at calving, then decreased significantly (P < 0.01). IgA and IgM on day 1 were 3.22 mg/ml and 5.22 mg/ml, respectively; both decreased during the first five days after calving. Values of IgA and IgM were higher than those reported in cows. SCC values, which were high at calving (500 000 per ml), reduced significantly (P < 0.01) on day 2, then decreased slightly until day 5 (180 000 per ml). At calving, macrophages were the most prominent cells in buffalo colostrum, followed by lymphocytes and neutrophils. Phagocytic activity was 23% at calving and reduced significantly (P < 0.01) to 14% on day 5. Phagocytic index was highest in the first colostrum, and then decreased non-significantly.     

Huizache (<Emphasis Type="Italic">Acacia farnesiana</Emphasis>) whole pods (flesh and seeds) as an alternative feed for sheep in Mexico

Two experiments were undertaken to evaluate the use of pods from Huizache (Acacia farnesiana), common in the arid and semiarid regions of Mexico, on the perfromance and apparent digestibility in Pelibuey Mexican hair growing ewe lambs. Twenty-four Pelibuey ewe lambs were used in the animal performance experiment, with a mean live weight of 14.91 ± 1.48 kg, randomnly allocated to three groups which received ad libitum for 77 days (11 weeks) experimental whole rations T0 with 0%, T12 with 12% or T24 with 24% inclusión of dried and ground Huizache pods. Dry matter intakes (g/kg ^0.75 daily) were 83, 95, 90 for T0, T12, and T24 respectively (P > 0.05). Mean daily live-weight gain was 90, 75, and 63 g/day for T0, T12, and T24 (P < 0.001). Nine Pelibuey ewe lambs were used to determine apparent digestibility in vivo of the experimental diets using a 3 × 3 latin square design repeated three times. There were differences in the digestibility of dry matter (P < 0.001), organic matter (P < 0.001), nitrogen (P < 0.031), neutral detergent fibre (P < 0.002), and acid detergent fibre (P < 0.001) being lower in T24. Huizache pods may be an alternative feed when included up to 12% of dry matter in the diets for sheep growing moderately.     

Ketosis in buffalo (Bubalus bubalis): clinical findings and the associated oxidative stress level

As little is known about the oxidant/antioxidant status in buffalo with ketosis, the present study was delineated to assess the oxidative stress level associated with clinical ketosis in water buffalo. A total of 91 parturient buffalo at smallholder farms were studied (61 suspected to be ketotic and 30 healthy). Clinical and biochemical investigations were carried out for each buffalo. Based on clinical findings and the level of beta-hydroxybutyrate (BHB), buffalo were allocated into ketotic (42), sublinical cases (19). Clinically, there was an association between clinical ketosis and anorexia (p < 0.001), constipation (p < 0.001), decreased milk yield (p < 0.001), ruminal stasis (p < 0.001), and loss of body condition (p < 0.01). Biochemically, in clinical ketosis compared with subclinical and control cases, there was a significant increase (p < 0.05) of BHB, malondialdehyde (MDA), nitric oxide (NO), aspartate aminotransferase (AST), L-alanine aminotransferase (ALT). However, there was a significant decrease of glucose, phosphorus, magnesium,total cholesterol and HDL-cholesterol. There was a positive correlation between BHB and MDA (r = 0.433), BHB and NO (r = 0.37), MDA and NO (r = 0.515), and Glucose and phosphorus(r = 0.521). However, there was a negative correlation between BHB and glucose (r = −0.341) and HDL and NO (r = −0.379). The result of the present study indicates that hyperketonemia in buffalo is associated with an increase of oxidative stress levels. Further studies need to be done on the efficacy of antioxidants as an ancillary treatment to relief the oxidative stress caused by ketosis.