Effects of Co‐culture of Cumulus Oocyte Complexes with Denuded Oocytes During In Vitro Maturation on the Developmental Competence of Cloned Bovine Embryos

This study evaluated the effects of co‐culture of immature cumulus oocyte complexes (COCs) with denuded immature oocytes (DO) during in vitro maturation on the developmental competence and quality of cloned bovine embryos. We demonstrated that developmental competence, judged by the blastocyst formation rate, was significantly higher in the co‐cultured somatic cell nuclear transfer (SCNT+DO, 37.1 ± 1.1%) group than that in the non‐co‐cultured somatic cell nuclear transfer (SCNT‐DO, 25.1 ± 0.9%) group and was very similar to that in the control IVF (IVF, 38.8 ± 2.8%) group. Moreover, the total cell number per blastocyst in the SCNT+DO group (101.7 ± 6.2) was higher than that in the SCNT‐DO group (81.7 ± 4.3), while still less than that in the IVF group (133.3 ± 6.0). Furthermore, our data showed that mRNA levels of the methylation‐related genes DNMT1 and DNMT3a in the SCNT+DO group were similar to that in the IVF group, while they were significantly higher in the SCNT‐DO group. Similarly, while the mRNA levels of the deacetylation‐related genes HDAC2 and HDAC3 were significantly higher in the SCNT‐DO group, they were comparable between the IVF and SCNT+DO groups. However, the mRNA levels of HDAC1 and DNMT3B were significantly higher in the SCNT+DO group than in the other groups. In conclusion, the present study demonstrated that co‐culture of COCs with DO improves the in vitro developmental competence and quality of cloned embryos, as evidenced by increased total cell number.     

Inheritance of histone H3 methylation in reprogramming of somatic nuclei following nuclear transfer

Successful cloning requires reprogramming of epigenetic information of the somatic nucleus to an embryonic state. However, the molecular mechanisms regarding epigenetic reprogramming of the somatic chromatin are unclear. Herein, we transferred NIH3T3 cell nuclei into enucleated mouse oocytes and evaluated the histone H3 dimethyl-lysine 4 (H3K4me2) dynamics by immunocytochemistry. A low level of H3K4me2 in the somatic chromatin was maintained in pseudo-pronuclei. Unlike in vitro fertilized (IVF) embryos, the methylation level of nuclear transfer (NT) embryos was significantly increased at the 8-cell stage. NT embryos showed lower H3K4me2 intensity than IVF embryos at the 2-cell stage, which is when the mouse embryonic genome is activated. Moreover, the H3K4me2 signal was weak in the recloned embryos derived from single blastomeres of the NT embryos, whereas it was intense in those from IVF embryos. Two imprinted genes, U2afbp-rs and Xist, were abnormally transcribed in cloned embryos compared with IVF embryos, and this was partly correlated to the H3K4me2 level. Our results suggest that abnormal reprogramming of epigenetic markers such as histone acetylation and methylation may lead to dysregualtion of gene expression in cloned embryos.     

DNA methylation status of bovine blastocyst embryos obtained from various procedures

DNA methylation is an important factor for the regulation of gene expression in early embryos. It is well known that the satellite I sequence is more heavily methylated in bovine somatic cell nuclear transfer (NT-SC) embryos than in embryos derived from in vitro fertilization (IVF). However, the methylation status of bovine embryos obtained by other procedures is not well known. To clarify DNA methylation levels of bovine embryos obtained from various procedures, we examined satellite I sequences in bovine blastocyst (BC) embryos derived from NT-SC, NT using embryonic blastomeres (NT-EM), in vivo (Vivo), IVF and parthenogenetic treatment (PA). Furthermore, in order to evaluate the efficacy of DNA demethylation by the NT procedure, we determined the DNA methylation levels in bovine embryos in which NT was recapitulated (Re-NT). Although the DNA methylation levels in the NT-SC embryos were higher than those in the other embryos, the NT-EM embryos exhibited lower DNA methylation levels. The satellite I sequence in the NT-SC embryos was more demethylated than that in the donor cells. Although the DNA methylation level in the individual NT-SC embryos showed variation, the full-term developmental efficacy of these embryos were not different. These findings suggest that the methylation level of the satellite I sequence at the BC stage is not related to the abnormalities of bovine embryos produced by NT-SC. There was no difference in methylation levels between Re-NT and NT-SC embryos. Our results indicated that the DNA methylation status differed among embryos produced by various methods and that at least some of the demethylation of the donor cell genome occurred in the recipient cytoplast after NT-SC, but the demethylation ability of the NT procedure was noted in the first NT but not in the second NT.     

Suberoylanilide hydroxamic acid,a novel histone deacetylase inhibitor,improves the development and acetylation level of miniature porcine handmade cloning embryos

Histone deacetylase inhibitors (HDACis) can change the histone acetylation and significantly enhance the developmental competence of the pre‐implantation SCNT embryo. To select a proper histone deacetylase inhibitor to improve the success rate and potentially developmental ability of handmade cloning (HMC) embryos of miniature porcine, we compared the effect of two histone deacetylase inhibitors (SAHA vs. VPA) on HMC embryo development, their histone acetylation level and the expression level of relevant genes. The blastocyst rate and number of blastocyst cells of HMC embryos treated with SAHA (SAHA‐HMC) or VPA (VPA‐HMC) were significantly higher than those of control (Control‐HMC), respectively, but there were no significant difference between SAHA‐HMC and VPA‐HMC groups. In addition, the acetylation level (AcH4K8) of Control‐HMC and VPA‐HMC embryos at the blastocyst stage, respectively, was significantly lower than that of in vitro fertilized (IVF) and SAHA‐HMC embryos. However, the acetylation H4K8 of the blastocysts had no significant difference between SAHA‐HMC and the IVF groups. The SAHA‐HMC blastocysts indicated comparative expression levels of Oct4 and HDAC1 (histone deacetyltransferase gene) with those of IVF blastocysts. In contrast, the expression levels of Oct4 were lower and those of HDAC1 were higher in the VPA‐HMC and Control‐HMC blastocysts, respectively, compared to those of the IVF blastocysts. Our results demonstrated that the HMC embryos treated by SAHA could promote the pre‐implantation development and increase the levels of histone H4K8 acetylation and the expression of the OCT4 gene, yet decrease the expression of the HDAC1 gene to the comparable level of the IVF embryos. Our results proved that SAHA may be a better histone deacetylase inhibitor for porcine HMC compared to VPA, and furthermore, it may indicate that SAHA can effectively correct the abnormal histone acetylation during the HMC embryo development and subsequently improve the full‐term developmental potential of the HMC embryos after embryo transplantation.     

Effect of histone acetylation modification with sodium butyrate, a histone deacetylase inhibitor, on cell cycle, apoptosis, ploidy and gene expression in porcine fetal fibroblasts

The present study evaluated the effective dose of sodium butyrate (NaB), a histone deacetylase (HDAC) inhibitor, for determination of the level of enhancement of histone acetylation in porcine fetal fibroblasts (PFFs) based on their morphology, growth, apoptosis and cell cycle status. Cells were analyzed for their histone acetylation levels at H3, H4 and H2A and expression of genes related to histone deacetylation (HDAC1, HDAC2 and HDAC3), pro-apoptosis (Bax and Bak) and anti-apoptosis (Bcl-2). PFFs at passage 3-4 were cultured with 0, 0.5, 1.0, 2.0 and 3.0 mM NaB for 96 h. NaB inhibited cell proliferation at all tested concentrations in a dose-dependent manner. However, there was slow cell growth for PFFs treated with 2.0 and 3.0 mM NaB compared with those of untreated PFFs and those treated with other lower concentrations (0.5 and 1.0 mM). More than 85% of the cells that were untreated or treated with 0.5 or 1.0 mM NaB had intact membranes, whereas, approximately 30% of the cells treated with 2.0 or 3.0 mM NaB had increased cell sizes and a more flattened and elongated appearance. NaB induced apoptosis in a dose-dependent manner; the rates of apoptosis were 2.5 +/- 0.4% for 1.0 mM NaB, 7.6 +/- 1.1% for 2.0 mM NaB and 11.2 +/- 1.4% for 3.0 mM NaB. The chromosomal sets of PFFs treated with 0.5 and 1.0 mM NaB were normal, whereas a lower proportion of PFFs treated with 2.0 and 3.0 mM were classified as normal. NaB at 0.5 and 1.0 mM showed little effect on cell cycle. However, 2.0 and 3.0 mM resulted in an increased cell population at the G(0)/G(1) phase. Increased NaB concentrations led to elevated acetylation of H3, H4 and H2A. NaB altered the expression of histone deacetylation and apoptosis-related genes. In conclusion, 1.0 mM NaB induced histone hyperacetylation in the PFFs and produced less deleterious effects than other concentrations; these PFFs might serve as suitable donors for porcine somatic cell nuclear transfer (SCNT).     

Extended exposure to trichostatin A after activation alters the expression of genes important for early development in nuclear transfer murine embryos

The low viability of embryos reconstructed by somatic cell nuclear transfer (SCNT) is believed to be associated with epigenetic modification errors, and reduction of those errors may improve the viability of SCNT embryos. The present study shows the effect of trichostatin A (TSA), a strong inhibitor of histone deacetylase, on the development of murine SCNT embryos. After enucleation and nuclear injection, reconstructed murine oocytes were activated with or without TSA for 6 hr (TSA-6 hr). After activation, TSA treatment was extended to 3 hr (TSA-9 hr), 5 hr (TSA-11 hr) and 18 hr (TSA-24 hr) during culture. As a result, the SCNT embryos in the TSA-11 hr group showed a remarkably higher blastocyst rate (21.1%) when compared with the nontreated embryos (3.4%), while the concentration of TSA did not significantly affect embryonic development. The expressions of histone deacetylase (HDAC1 and HDAC2) and DNA methylation (DNMT3a and DNMT3b) genes decreased in the TSA-11 hr and TSA-24 hr groups, while there was an increase in the expression of histone acetyltransferase (P300 and CBP), pluripotency (OCT4 and NANOG) and embryonic growth/trophectoderm formation (FGF4)-related genes in the same groups. The expression of CDX2, a critical gene for trophectoderm formation was upregulated only in the TSA-24 hr group. Our results show that TSA treatment during the peri- and postactivation period improves the development of reconstructed murine embryos, and this observation may be explained by enhanced epigenetic modification of somatic cells caused by TSA-induced hyperacetylation, demethylation and upregulation of pluripotency and embryonic growth after SCNT.     

Buffalo SCNT embryos exhibit abnormal gene expression of ERK/MAPK pathway and DNA methylation

Inhibition of ERK/MAPK pathway has been shown to decrease DNA methylation via down‐regulation of DNA methyltransferases (DNMTs) in several studies suggesting that this pathway plays an important role in regulation of DNA methylation. We examined the relative expression level of seven important genes related to ERK/MAPK pathway and DNMTs (DNMT1, DNMT3a and DNMT3b) by quantitative real‐time PCR in buffalo blastocysts produced by Hand‐made cloning and compared it with that in blastocyst‐stage embryos produced by in vitro fertilization (IVF). The expression level of six of seven genes related to ERK/MAPK pathway examined i.e., p21RAS, RAF1, AKT1, ERK2, PIK3R2 and c‐Myc was significantly higher (p < 0.05) in cloned than in IVF embryos. However, the expression level of FOS was lower (p < 0.005) in cloned than in IVF embryos. The relative expression level of DNMT3a and DNMT3b but not that of DNMT1 was significantly higher (p < 0.05) in cloned than in IVF embryos. These results indicate that the cloned embryos exhibit an abnormal expression of several important genes related to ERK/MAPK pathway and DNMTs. Although a direct link between ERK/MAPK pathway and DNMTs was not examined in the present study, it can be speculated that ERK/MAPK pathway may have a role in regulating the expression of DNMTs in embryos, as also observed in other tissues.     

Defined system for in vitro production of porcine embryos using a single basic medium

We have previously indicated that porcine blastocysts can be produced by in vitro fertilization (IVF) and culture (IVC) in chemically defined porcine gamete medium (PGM) and porcine zygote medium (PZM)-5, respectively, In the present study, the effects of basic media and macromolecular components on in vitro maturation (IVM) were investigated to develop a defined system for in vitro embryo production using a single basic medium through IVM, IVF and IVC. Porcine immature oocytes were matured in porcine oocyte medium (POM) or modified North Carolina State University (mNCSU) 37, which were supplemented with either 10% (v/v) porcine follicular fluid (pFF) or 3 mg/ml polyvinyl alcohol (PVA) as a macromolecular component (designated POM+pFF, POM+PVA, mNCSU37+pFF and mNCSU37+PVA). In the maturation with mNCSU37+PVA, the percentages of oocytes that reached the metaphase II stages were significantly lower than those in the other treatments. Following IVM with the above media, oocytes were treated with an electrical stimulus and cycloheximide for parthenogenetic activation and were cultured in PZM-5 for 5 days. The rates of cleavage and blastocyst formation of parthenogenetic oocytes were significantly lowered for maturation with mNCSU37+PVA compared with the other treatments, while there were no significant differences in the total numbers of cells in blastocysts among the treatments. Following IVF and IVC, the rates of penetration, male pronucleus formation, cleavage and blastocyst formation were significantly lower when oocytes were matured in mNCSU37+PVA than in other maturation media. The normal fertilization rate was significantly higher in POM+PVA compared with the other treatments, although the total number of cells in blastocysts was reduced with the addition of PVA to both POM and mNCSU37 compared with pFF supplementation. These results demonstrate that porcine blastocysts can be produced by the defined system using a single basic medium.     

Ovarian evaluation of Girolando (Holstein × Gir) heifers submitted to a GnRH–PGF2α–GnRH protocol in the dry or rainy seasons in the tropical savannah

The Girolando breed is used in pasture-based dairy production systems in Brazil to associate the high production of Bos taurus to the rusticity and thermal adaptation of Bos indicus. This study was designed to evaluate the physiological response to a gonadotropin-releasing hormone (GnRH)–prostaglandin F_2α (PGF_2α)–GnRH protocol to synchronize the ovulation in 40 Girolando heifers of a pasture-based dairy production system and its relationships with the temperature and humidity index (THI) during the dry (DS) and rainy season (RS) in the tropical savannah—Brazil's cerrado biome. Responses were characterized by follicular and corpus luteum number and diameter, ovulation (D9), and pregnancy rates after first AI. Total follicle number (8.1?±?0.3?×?8.8?±?0.3), D9 ovulatory follicle diameter (11.9?±?0.4?×?10.1?±?0.4 mm), corpus luteum diameter (8.6?±?1.3?×?3.9?±?1.5 mm), corpus luteum score (3.7?±?0.8?×?1.8?±?1.0), corpus luteum diameter after AI (9.6?±?1.6?×?3.9?±?1.5 mm), and corpus luteum score after AI (3.2?±?0.4?×?0.9?±?0.6) in DS and RS differed (P?<?0.01). D9 ovulation rate was 40 % (DS) and 20 % (RS), without differences (P?>?0.05). Pregnancy rate was 45 % (DS) and 11 % (RS), with differences (P?<?0.01). THI differed between DS and RS (P?<?0.01). THI may interfere in the follicular and luteal dynamics and in the response of Girolando heifers to the GnRH–PGF_2α protocol in the tropical savannah, thus reducing the chances of pregnancy at the first artificial insemination.     

m‐carboxycinnamic acid bishydroxamide improves developmental competence,reduces apoptosis and alters epigenetic status and gene expression pattern in cloned buffalo (Bubalus bubalis) embryos

Incomplete or aberrant reprogramming of nuclear genome is one of the major problems in somatic cell nuclear transfer. In this study, we studied the effect of histone deacetylase inhibitor m‐carboxycinnamic acid bishydroxamide (CBHA) on in vitro development of buffalo embryos produced by Hand‐made cloning. Cloned embryos were treated with CBHA (0, 5, 10, 20 or 50 μM) for 10 hr from the start of reconstruction till activation. At 10 μM, but not at other concentrations examined, CBHA increased (p < .05) the blastocyst rate (63.77 ± 3.97% vs 48.63 ± 3.55%) and reduced (p < .05) the apoptotic index of the cloned blastocysts (8.91 ± 1.94 vs 4.36 ± 1.08) compared to untreated controls, to levels similar to those in IVF blastocysts (4.78 ± 0.74). CBHA treatment, at all the concentrations examined, increased (p < .05) the global level of H3K9ac in cloned blastocysts than in untreated controls to that observed in IVF blastocysts. Treatment with CBHA (10 μM) decreased (p < .05) the global level of H3K27me3 in cloned blastocysts than in untreated controls but it was still higher (p < .05) than in IVF blastocysts. CBHA (10 μM) treatment increased (p < .05) the relative expression level of pluripotency‐related genes OCT‐4 and NANOG, and anti‐apoptotic gene BCL‐XL, and decreased (p < .05) that of pro‐apoptotic gene BAX than in untreated controls but did not affect the relative expression level of apoptosis‐related genes p53 and CASPASE3 and epigenetics‐related genes DNMT1, DNMT3a and HDAC1. These results suggest that treatment of cloned embryos with 10 μM CBHA improves the blastocyst rate, reduces the level of apoptosis and alters the epigenetic status and gene expression pattern.     

Production potentials of native chickens (Gallus gallus domesticus L.) of Western Visayas, Philippines

Poultry genetic resources diversity serves as a reservoir of genes adaptable to local conditions, production, resource utilization, and production that are compatible with consumer preferences for meat and egg products. This study was, therefore, conducted to determine the production potentials of native chickens in Western Visayas, Philippines. A total of 270 raisers were interviewed and 810 chickens were characterized. These numbers were divided equally among the provinces of Aklan, Antique, Capiz, Guimaras, Iloilo, and Negros Occidental. The data collected were analyzed using one-way analysis of variance and significant differences were determined using the Duncan’s Multiple Range Test. Results show that native chickens from Aklan had the highest (P?<?0.01) hatching percentage (93.10?±?1.67 %) and oldest (P?<?0.01) slaughter/marketing age (23.56?±?1.03 weeks). Those from Guimaras had the biggest (P?<?0.01) egg (42.69?±?0.87 g) and heaviest slaughter/marketing weight (920.00?±?22.72 to 924.44?±?23.01 g); the shortest (P?<?0.01) clutch interval (77.87?±?4.14 days) was observed from those in Iloilo. Those from Antique had the highest (P?<?0.01) survival rate from 1 day old to a slaughter/marketing age of 52.48?±?1.83 %. The adult live weight ranged from 1.27?±?0.04 to 1.62?±?0.04 kg with those from Aklan as the lowest (P?<?0.01) while those from Guimaras were the highest. Thus, the observed variations in reproductive and growth performances of native chickens from this region expressed their potentials for genetic improvement toward production.     

Epigenetic characteristics of cloned and in vitro-fertilized swamp buffalo (Bubalus bubalis) embryos

Swamp buffalos are becoming endangered due to reproductive inefficiencies. This is of concern because many countries depend heavily on their products. Somatic cell nuclear transfer (SCNT) is a potential strategy for preserving endangered species. To date, SCNT in swamp buffalo has succeeded in the creation of blastocyst embryos. However, development to term of SCNT swamp buffalos is extremely limited, and only 1 live birth has been reported. An abnormal epigenetic mechanism is suspected to be the cause of developmental failure, as is also seen in other species. The DNA methylation and histone acetylation are key players in epigenetic modification and display marked variability during embryonic preimplantation development. Knowledge of epigenetic modifications will aid in solving the developmental problems of SCNT embryos and improving reproductive technology in the swamp buffalo. The objective of this study was to determine the relationship between preimplantation embryonic development and 2 epigenetic patterns, global DNA methylation and histone acetylation, in SCNT and in vitro-fertilized (IVF) swamp buffalo embryos. In addition, we examined the correlations between those 2 mechanisms in the SCNT and IVF swamp buffalo embryos throughout the developmental stages using double immunostaining and quantification of the emission intensities using confocal microscopy. We discovered an aberrant methylation pattern in early preimplantation-stage swamp buffalo SCNT embryos. In addition, greater variability in the DNA methylation levels among nuclei within SCNT embryos was discovered. Hyperacetylation was also observed in SCNT embryos compared with IVF embryos at the 4- and 8-cell stages (P < 0.05). Dynamic changes and interplay between these 2 epigenetic mechanisms could be crucial for embryonic development during the early preimplantation period. The aberrancies uncovered here may contribute to the low efficiency of SCNT.     

Pharmacokinetics of lincomycin following single intravenous administration in buffalo calves

Lincomycin 10 mg kg^?1, IV in buffalo calves followed two-compartment open model with high distribution rate constant α (11.2?±?0.42 h^?1) and K ₁₂/K ₂₁ ratio (4.40?±?0.10). Distribution half-life was 0.06?±?0.01 h and AUC was 41.6?±?1.73 μg mL^?1 h. Large Vd_area (1.15?±?0.03 L kg^?1) indicated good distribution of lincomycin in various body fluids and tissues. Peak plasma level of lincomycin (71.8?±?1.83 μg mL^?1) was observed at 1 min as expected by IV route. The elimination half-life and MRT of lincomycin were short (3.30?±?0.08 and 4.32?±?0.11 h, respectively). Lincomycin 10 mg kg^?1 IV at 12-h interval would be sufficient to maintain T?>?MIC above 60 % for bacteria with minimum inhibitory concentrations (MIC) values ≤1.6 μg mL^?1. Favourable pharmacokinetic profile in buffalo calves and a convenient dosing interval suggest that lincomycin may be an appropriate antibacterial in buffalo species for gram-positive and anaerobic bacterial pathogens susceptible to lincomycin.     

Ultrasonographic studies on ovarian dynamics and associated estrus manifestations of jennies under controlled management, Ethiopia

A serial ultrasonographic study was conducted on nine jennies aged 5–15 years from January to April 2008 with the objective of studying ovarian follicular dynamics and estrus manifestations under controlled management. Ovarian follicular activity was determined from the number and size distribution of follicles, length of interovulatory interval (IOI), growth rate of preovulatory follicles, diameter of follicles at the onset of estrus, and incidence of ovulation. Estrus manifestations were characterized using length of estrus and estrous cycle. The mean (±SD) number of follicle detected per ovary was 5.45?±?2.3 (range, 1–16) with sizes ranging from 2.9 to 44 mm. The mean (±SD) size of follicle encountered at the onset of estrus was 25.9?±?3.7 mm (range, 20.9–34.4) while that of the preovulatory follicles at ?1 day before ovulation was 36.81?±?3.78 mm. The mean (±SD) IOI, estrus, and estrous cycle length were 25.4?±?3.6, 7.9?±?2.9, and 24.2?±?7.4 days, respectively. The mean (±SD) growth rate of the preovulatory follicle after the day of divergence was 1.9?±?0.3 mm/day. Serum progesterone profile followed the same patterns of ovarian dynamics with maximum values being detected during midluteal phase. Serum progesterone assay revealed blood progesterone profiles of <1.0 ng/ml during estrus and up to 11 ng/ml during midluteal phase with a pattern following follicular dynamics. Body condition of the study jennies steadily increased and was positively correlated (r?=?0.52, p?<?0.001) with the diameter of the preovulatory follicle. In conclusion, the ultrasonic evaluation has revealed that follicular dynamics of jennies were generally related with body condition which might have been influenced by the type of management.     

Estrone sulfate and progesterone profiles during late gestation in recipient cows transferred embryos produced by nuclear transfer and in vitro fertilization

The aim of present experiment was to evaluate the plasma concentrations of estrone sulfate (E(1)S) and progesterone (P(4)) during late gestation in recipient cows transferred embryos produced by nuclear transfer (NT) and in vitro fertilization (IVF). Blood samples were collected from recipients transferred embryos produced by NT (n=9) and IVF (n=13) at 160, 220, 240, 260 and 270 d of gestation and then at 5 d intervals until parturition. Plasma samples were analyzed for E(1)S and P(4) by ELISA. One NT and three IVF cows aborted between days 220 and 260 of gestation. Two NT and one IVF cow had prolonged gestation (over 290 d). One IVF cow had an overweight fetus (50 kg) after abortion (257 d). The patterns of changes in the concentrations of E(1)S during late gestation in the NT and IVF cows were almost identical. The NT and IVF cows that aborted had prolonged gestation and much higher E(1)S concentrations than the average. One NT cow aborted after 220 d of gestation and had a sudden high increase in its E(1)S concentration from 160 d to 220 d of gestation. The NT and IVF cows that had prolonged gestation also had significantly higher (P<0.05) P(4) concentrations than the average. These results raise the possibility that the E(1)S and P(4) profiles can be used to monitor some late gestational problems, such as higher birth weight, abortion and prolonged gestation.     

Prospects of raising Sahiwal cow calves for veal production under tropical environment

The objective of present study was to evaluate the growth potential of Sahiwal calves for veal production on whole milk or whole milk and milk replacer combined in a ratio of 50:50 (MMR). For this purpose, 48 Sahiwal calves (both male and female) were assigned to four dietary treatments having 12 animals/treatment. Calves in the treatments A and B were offered whole milk at 15 or 20 % of their body weight (BW), respectively, up to day 84 adjusted on weekly basis. The calves in treatments C and D received the same amount of milk as in treatments A and B until day 21, respectively, after which 50 % of the milk offered was replaced with a blend of chickpea (Cicer arietinum) flour and vegetable (corn) oil mixed in water (MR) until day 84. The constituted MR had 3.1, 2.8, and 14.3 %, CP, EE, and DM, respectively. The growth and intake data were analyzed using repeated measures analysis, with MIXED Procedures of SAS in a 2?×?2 factorial design. The two factors were feeding level and feeding source. Calves offered whole milk grew faster (P?<?0.05) and had greater weaning weights (P?<?0.05) than those offered MMR (606.4?±?18.1 vs 331.3?±?18.1 g/day and 70.4?±?1.5 vs 47.8?±?1.5 kg, respectively). Greatest daily BW gain (656?±?26 g/day) and weaning weight (74.6?±?2.1 kg) were observed in calves given treatment B while the lowest production cost/kg of BW gain (≈US$3.6?±?0.2) was observed in calves given treatment A. The daily BW gain of calves fed milk ad libitum was 716?±?40 and 836?±?40 g/day, during 5–8 and 9–12 weeks, respectively. The number of days calves exhibited scours was higher in calves offered MMR than those offered whole milk. Replacement of 50 % milk with a blend of chickpea flour and vegetable oil, as an alternative to milk replacer, did not support growth equivalent to whole milk and was not effective in reducing feeding cost during the weaning period. Sahiwal calves may have promise for being raised for veal production under tropical environments.