Nucleolus organizer regions and heterochromatin in the zebu (Bos indicus L.)

Summary Ag-NOR staining and a counterstain enhanced fluorescence technique (chromomycin A₃/distamycin A/DAPI-staining = CDD-method) and G-banding, respectively, have been applied to the zebu (Bos indicus L.) chromosomes. The nucleolus organizer regions (NORs) were found in the telomeric regions of chromosomes nos. 2, 3, 4, 11, and 28. CDD staining led to a well-defined R-banding pattern along the chromosome arms and to the visualization of centric heterochromatic bands of variable sizes.     

Identical marker alleles in Podolic cattle (Bos taurus) and Indian zebu (Bos indicus)

In the context of biochemical marker research and in order to add new information on native breeds, the present work focuses on a local Southern Italy cattle, namely Italian Podolic. We provide the complete structural characterisation of alpha-lactalbumins and beta-globin chains isolated from Podolic cattle (Bos taurus). Given the unavailability of the complete sequence for alpha-lactalbumin A of taurine cattle in the literature, we intended to check its structure in order to ascertain the absence of any possible silent mutation. Screening the Podolic cattle, we found a new beta-globin variant not detectable by conventional methods. The presence of such a new variant might be helpful in the study of the Podolic population genetic structure and for a better knowledge of the gene pool per se, and in comparison with the other breeds. Structural analyses showed that the new beta-globin Podolic variant exhibited the same sequence as beta-globin Azebu. The alpha-lactalbumin A was the same as that isolated from zebu cattle (Bos indicus). The results are discussed in relation to the possible involvement of the two markers in the debate on the origin of the Podolic breed.     

Peripheral plasma progesterone concentration in zebu (Bos indicus) cows during pregnancy

Plasma progesterone concentrations were determined weekly during gestation averaging 283 +/- 2 d in Ethiopian zebu (Bos indicus) cows. Mean progesterone levels increased from 0.2 +/- 0.1 ng/ml at oestrus (service) to 3.1 +/- 1.6 ng/ml on d 7 and 8.1 +/- 2.1 ng/ml on d 21. Progesterone levels remained elevated throughout pregnancy. Hormone concentration differed significantly between cows (P less than 0.001) and with the wk of pregnancy (P less than 0.05); it tended to be higher during the last trimester of pregnancy. Mean levels dropped sharply to below 1 ng/ml during the last wk of pregnancy with considerable variation (C.V. = 39 to 63%) among cows. These data indicate that pregnancy in Ethiopian zebu cows can be reliably diagnosed by determining circulatory plasma progesterone levels.     

Studies on purine turnover in the camel (Camelus dromedarius) and zebu (Bos indicus)

Significantly higher hypoxanthine over uric acid ratios were found in camel plasma and urine, with respect to those of zebu. Enzyme levels of purine catabolism were markedly lower in camel than in zebu liver. Oxidation of hypoxanthine appears to be the limiting step of purine metabolism in camel liver. Any hepatic hypoxanthine appears to be actively converted into IMP in camel liver, rather than oxidized to uric acid.     

Ovarian follicular dynamics in suckled zebu (Bos indicus) cows monitored by real time ultrasonography

The pattern of follicular growth was studied in 17 suckled zebu cows with average body condition and under extensive management in a tropical environment (23 degrees C, 78% humidity; 2200 mm annual rainfall; 1000 m altitude). The study covered the period from parturition to weaning at 12 months postpartum (PP). Data were collected by transrectal ultrasonography (7.5 MHz) at 48 h intervals, and progesterone (P4) measurements were performed by RIA. The sequential development of ovarian follicles greater than 4 mm was followed until regression or ovulation. Ovarian activity as characterized by growth and regression of follicles of 4 to 6 mm, with sporadic dominance, and a long interdominance interval was observed in every cow and from as early as 26 +/- 2 days PP. This follicular pattern was highly variable during the first 6 months: cows presented 2 to 20 follicular waves (FW) in which a dominant follicle (DF) grew to 8 +/- 1 mm with daily growth rates of 1.1 +/- 0.5 mm/day. The duration of dominance varied from 2 to 8 days and the interdominance time interval was 0 (overlapped waves) to 60 days. Neither behavioural oestrus nor ovulation was observed during this period. From 6 to 12 months PP, cows presented 7 to 20 FW, some with ovulation and/or corpora lutea (CL) formation. The ovulation was preceded by oestrus in some cases (43%). The mean (+/- sem) diameter of DF was 9 +/- 2.7 mm, their mean growth rate 1.4 +/- 0.2 mm/day, their duration of dominance was 2 to 8 days and the interdominance interval was 0 to 14 days. Progesterone concentrations (P4) from 1.0 to 13 ng/ml were found when a CL was present. Once cyclicity re-commenced at 217 to 278 days PP, the cows presented either normal (21 +/- 3 days), short (10 +/- 2 days), or long (50 +/- 4 days) cycles. The resumption of cyclicity was characterized by an increased frequency of emerging follicular waves. Under the conditions of this study, the suckled Bos indicus cows re-commenced ovarian follicular activity as early as described in B. taurus breeds, but the establishment of cyclicity was substantially later. These data add further to the panorama of postpartum reproductive physiology in tropical cattle.     

Effects of lowered temperatures and media on short-term preservation of zebu (Bos indicus) preantral ovarian follicles

The maintenance of follicle quality during the transportation of ovaries is essential for the successful cryopreservation and in vitro development of preantral follicles. The objective of this study was to evaluate the effect of cooling ovarian tissue on the conservation of zebu cow preantral follicles. Ovarian pieces were immersed in saline or coconut water (CW) solutions and maintained at 4 or 20 degrees C for 6, 12, or 18 h. Preantral follicles were evaluated by histology and transmission electron microscopy. Storage of ovarian pieces at 20 degrees C for 12 or 18 h significantly reduced the percentage of morphologically normal follicles compared to controls. In contrast, conservation at 4 degrees C for up to 18 h and at 20 degrees C for up to 6 h kept the percentage of normal follicles similar to controls. However, the type of solution that the ovaries were immersed in had little effect on the results. Decreased cellular metabolism probably accounted for better preservation of preantral follicles at 4 degrees C. In conclusion, zebu cow ovaries were successfully stored at 4 degrees C for up to 18 h with no morphological damage to preantral follicles. However, at 20 degrees C, ovaries could only be stored for 6 h.     

Characterization of pregnancy-associated glycoproteins extracted from zebu (Bos indicus) placentas removed at different gestational periods

In the present work, two biochemical approaches were used to characterize PAGs isolated from Bos indicus fetal cotyledons removed at different gestational ages. The first procedure included acidic and ammonium sulfate precipitations, anion and cation exchange chromatographies and the second included pepstatin-agarose affinity chromatography. A bovine PAG radioimmunoassay was used to monitor the immunoreactivity throughout the isolation procedures. The most immunoreactive fractions issued from cation exchange and affinity chromatographies were analyzed by SDS-PAGE and Western blotting, before transfer to a polyvinylidene difluoride (PVDF) membrane for NH2-microsequence determination. Use SDS-PAGE and Western blotting, different isoforms of PAG with apparent molecular masses of 51 to 69 kDa and isoelectric points varying from 4.4 to 6.7 were identified in the placentas from different gestational ages. N-terminal microsequencing (10 to 25 aa long) indicates the expression of one single terminal amino acid sequence in the Bos indicus placenta, which is 100% identical to the bovine PAG-1.     

Effect of different cryoprotectants on the structural preservation of follicles in frozen zebu bovine (Bos indicus) ovarian tissue

Cryopreservation of ovarian tissue is a new and promising technique for germ-line storage. The objective of this study was to evaluate the effect of four cryoprotectants (at two concentrations each) on the preservation of zebu bovine preantral follicles after ovarian cryostorage. Strips of ovarian cortex were cryopreserved using glycerol (GLY; 10 or 20%), ethylene glycol (EG), propanediol (PROH) or dimethylsulphoxide (DMSO; 1.5 or 3M). In addition, a toxicity test was performed for each cryoprotectant by exposing the ovarian tissue to them without freezing. Tissues were analyzed by histology and transmission electron microscopy. Ovarian tissue frozen in either concentration of DMSO or PROH or in 10% GLY retained a higher percentage of morphologically normal follicles (73-88%) than tissue frozen in 20% GLY or in either concentration of EG (16-52%). In the toxicity test, exposure of tissues to DMSO, PROH or GLY resulted in higher percentages of normal follicles (80-97%) than exposure to EG (49%). Electron microscopy revealed damage to the ultrastructure of follicles frozen in 10% GLY, while follicles cryopreserved in DMSO and PROH at either concentration exhibited normal ultrastructure. In conclusion, DMSO and PROH were the most effective cryoprotectants for zebu ovarian tissue, preserving the structural integrity of somatic and reproductive cells within the ovary.     

Serum progesterone profiles of zebu cattle (Bos indicus) in relationship to conception and repeat breeding after artificial insemination

Sixty-four zebu cows, comprised mainly of Bunaji cattle aged between 3 and 7 years old, were involved in an intensive artificial insemination programme. The study was conducted during the breeding period (May-October), and cows were inseminated with freshly prepared Friesian semen when they stood to be mounted by the vasectomised bull or herdmates. Fertility was measured by serum progesterone (P(4)) concentrations and pregnancy diagnosis by rectal examination post-insemination. From the records of oestrus detection and P(4) profiles of cattle following breeding, 39 oestrous cycle lengths were classified into short, normal and long cycles. The mean duration for short, normal and long oestrous cycles were 15.6+/-2.0, 21.5+/-1.5 and 29.5+/-2.5 days, respectively. Mean inter-oestrus intervals between the treatment groups were not different. The percentage frequency distribution was 48.7% for normal oestrous cycles. The short oestrous cycle lengths of 11-17 days were observed in 12.8% of the cases, while 38.5% of the cases of oestrus returns had long luteal phases with oestrous cycle lengths of 26-32 days.Out of the 64 cows, 48.4% conceived to the first breeding while 18.8% conceived to the second breeding. Five and nine cows became acyclic after the first and second breeding, respectively, constituting 22. 0%. Cows that displayed irregular oestrous cycles (repeat breeders) constituted 10.9%. Average number of services per conception was 1.3. Serum P(4) is of practical value in monitoring ovarian activity in cattle and in the identification of acyclic and repeat breeder cows. Repeat breeder cows could be investigated by a combination of radioimmunoassay and clinical examination of the ovaries and reproductive tract. In cattle management, it is economical and profitable to diagnose pregnancy early after insemination so that cows which fail to conceive may be rebred.     

Fertility of Bos indicus and Bos indicus x Bos taurus crossbreed cattle after estrus synchronization

Ninety-five cows (79 Boran and 16 Boran-Brahman crossbreeds) and 107 heifers (55 Boran and 52 Boran x Friesian F1 crossbreeds) were used to determine estrus response, estrus response interval and pregnancy rate following synchronization with prostaglandin (PGF(2)alpha), a progesterone-releasing intravaginal device (PRID) and Synchro-mate B (SMB). The proportion of cattle responding to synchronization treatment was 62.5, 43.5 and 57.7% for cows and 85.7, 68.0 and 81.5% for heifers using PGF(2)alpha, PRID and SMB, respectively. The overall mean response was 59 and 81.8% for cows and heifers, respectively. The estrus response of the control animals over a 45-d breeding period was 72.7 and 90% for cows and heifers, respectively. The estrus response interval for cows was 31.8, 22.1 and 18.0 h and it was 51.1, 38.0 and 21.6 h for heifers with PGF(2)alpha, PRID and SMB treatment, respectively. Mean pregnancy rate for cows was 50.0, 34.8, 46.2 and 68.8% and for heifers it was 60.7, 40.0, 55.6 and 77.8% in the PGF(2)alpha, PRID, SMB and control groups, respectively. Based on these findings, it was concluded that both PGF(2)alpha and SMB produce a satisfactory estrus response and pregnancy rate in the cattle studied.     

Sexual development in early- and late-maturing Bos indicus and Bos indicus x Bos taurus crossbred bulls in Brazil

Two experiments were conducted to evaluate sexual development in early- and late-maturing Nelore (Bos indicus) and Canchim (3/8 Bos indicus x 5/8 Bos taurus crossbred) bulls and to determine predictors of sexual precocity, and pubertal and maturity status. In Experiment 1, 12 Nelore bulls where examined from 300 to 900 days of age. Puberty was characterized by an ejaculate containing > or =50 million sperm with > or =10% motile sperm, and maturity by an ejaculate containing > or =70% morphologically normal sperm. In Experiment 2, 28 Canchim bulls where examined from 295 to 488 days of age and puberty was characterized by an ejaculate containing > or =30% motile sperm. In both experiments, bulls were classified as early- or late-maturing based on age at puberty. Early-maturing bulls were younger (P < 0.05) than late-maturing bulls at puberty (527 days versus 673 days in Experiment 1 and 360 days versus 461 days in Experiment 2) and at maturity (660 days versus 768 days in Experiment 1). In general, early-maturing bulls were heavier and had greater scrotal circumference (SC), testes, and testicular vascular cone diameter than late-maturing bulls during the experimental period. Scrotal circumference adjusted for 365 days of age was a good predictor of sexual precocity; minimum yearling SC of 19 and 24 cm for Nelore and Canchim bulls, respectively, had the best predictive values. Early-maturing bulls were lighter and had smaller SC at puberty than late-maturing bulls; therefore, sexual precocity was not related to the attainment of a threshold body weight or testicular size earlier, but to lower thresholds in early-maturing bulls. When predictors of pubertal status were evaluated, SC had the best sensitivity/specificity relationship in Nelore bulls, and high sensitivity and specificity in Canchim bulls. When predictors of sexual maturity were evaluated in Nelore bulls, age, weight, and SC had similar sensitivity, specificity, and predictive values. At puberty, approximately 60% of the sperm present in the ejaculate were morphologically defective. Changes in semen quality after puberty in Nelore bulls were characterized by increased motility and proportion of morphologically normal sperm, with a decrease in the proportion of major sperm defects. In conclusion, early-maturing bulls were more developed in the pre-pubertal period and attained puberty at earlier stages of body and testicular development than late-maturing bulls. Yearling SC could be used to select bulls for sexual precocity and SC was the best predictor of pubertal status. Age, weight, and SC were equally good predictors of sexual maturity in B. indicus bulls.     

Microsatellite analysis of genetic population structure of zebu cattle (Bos indicus) breeds from north-western region of India

The present study aims to understand the existing genetic diversity and structure of six native cattle breeds (Rathi, Tharparkar, Nagori, Mewati, Gir, and Kankrej) adapted to the north-western arid and semi-arid region of India based on microsatellite loci. Various diversity estimates, mean number of alleles (12.84); effective number of alleles (5.02); gene diversity (0.769), and observed heterozygosity (0.667) reflected the existence of substantial within-breed diversity in all the investigated cattle breeds. Mean estimates of F-statistics: F(IT) = 0.144 ± 0.023, F(IS) = 0.071 ± 0.021, and F(ST) = 0.078 ± 0.014 were significantly different from zero (P < 0.05). The interbreed relationships indicated moderate level of breed differentiation between the six cattle breeds with least differentiation between Kankrej-Mewati pair. The phylogeny structuring further supported close grouping of Kankrej and Mewati breeds. Correspondence analysis plotted Rathi, Tharparkar, and Gir individuals into three separate areas of multivariate space; whereas, Kankrej, Mewati, and Nagori cattle showed low breed specific clustering. This reflected the existence of discrete genetic structure for Tharparkar, Rathi, and Gir, the prominent dairy breeds of the region; whereas, admixture was observed for Kankrej, Mewati, and Nagori individuals.     

Effect of early luteolysis in progesterone-based timed AI protocols in Bos indicus, Bos indicus x Bos taurus, and Bos taurus heifers

The objective of this study was to evaluate the effects of treatment with an intravaginal progesterone-releasing device (CIDR) and estradiol benzoate (EB) on follicular dynamics in Bos indicus (n=23), Bos taurus (n=25), and cross-bred (n=23) heifers. To assess the influence of reduced serum progesterone concentrations during 8 days of treatment with a progesterone-releasing device on follicular dynamics, half of the heifers received PGF at CIDR insertion (Day 0; 3 x 2 factorial design). Mean (+/-S.E.M.) serum progesterone concentrations during CIDR treatment varied (P<0.05) among genetic groups: B. indicus (5.4+/-0.1 ng/mL), B. taurus (3.3+/-0.0 ng/mL), and cross-bred (4.3+/-0.1 ng/mL). Maximum diameter of the dominant follicle (DF) was smaller (P<0.01) in B. indicus heifers (9.5+/-0.5 mm) than in cross-bred (12.3+/-0.4 mm) or B. taurus heifers (11.6+/-0.5 mm). B. indicus experienced lower (P<0.01) ovulation rate (39.1%) than did B. taurus (72.7%) and cross-bred (84.0%). Heifers treated with PGF on Day 0 had lower (P<0.05) serum progesterone concentrations during progesterone treatment. The PGF treatment on Day 0 increased (P<0.01) the diameter of the DF (11.9+/-0.4 mm vs. 10.5+/-0.4 mm). Moreover, greater (P=0.02) ovulation rates (78.8 vs. 54.0%) occurred in heifers treated with PGF on Day 0. In summary, B. indicus heifers had greater serum progesterone concentrations, smaller DF diameter, and a lower ovulation rate compared to B. taurus heifers. Prostaglandin treatment on the day of CIDR insertion reduced serum progesterone during treatment, and resulted in increased maximum DF diameter and ovulation rate.     

Influence of bull biostimulation, season and parity on resumption of ovarian activity of zebu (Bos indicus) cattle following parturition

A total of 135 postpartum suckled zebu cows were assigned randomly at calving to two treatments: cows exposed to vasectomised bulls (Mature Bull Exposure, MBE) and cows not exposed to bulls (No Bull Exposure, NBE) for a period of 150 days postpartum. This study was conducted to determine the influence of bull biostimulation, season and parity on postpartum reproductive performance of Bos indicus cattle. The trials were conducted in two seasons: cows calving in the dry season and cows calving in the rainy season. Cows with an increase in serum progesterone (P(4)) concentration of >1 ng/ml from the weekly blood samples were used to analyse the number of days from calving to the time of resumption of ovarian activity. The introduction of vasectomised bulls shortened the postpartum anoestrus in cattle following calving. Mean interval from calving to resumption of ovarian activity for the MBE cows was 71.7 days, which was significantly earlier following calving than the NBE cows with a duration of 77.8 days. By 60 to 80 days postpartum, the proportion of cows at resumption of postpartum activity for MBE cows was greater than for the NBE cows.Mean interval from calving to resumption of ovarian activity for cows that calved in the dry season was 71.3 days, which was significantly earlier than for cows that calved in the rainy season (78.6 days). At 60 days postpartum, the proportion of cows at resumption of postpartum ovarin activity for cows that calved in the dry season was greater than the cows that calved in the rainy season. Mean interval from parturition to resumption of ovarian activity for cows with three to five calvings was 65.1 days, which was significantly earlier than the value of 71. 2 days for cows with one to two calvings. By 60 to 80 days postpartum, the proportion of cows at onset of ovarian activity for cows with three to five calvings was greater than those cows with one to two calvings. Cows that calved in the dry season completed uterine involution by 24.4 days, which was significantly shorter than the duration of 26.5 days for cows that calved in the rainy season. Cows with three to five calvings completed uterine involution earlier than those with one to two calvings. It is concluded that bull-cow biostimulation influences reproductive activity in the cow possibly through olfactory cues (pheromones).     

Ciliate protozoa in the rumen of Kenyan zebu cattle, Bos taurus indicus, with the description of four new species

The composition of the fauna of rumen ciliates in zebu cattle in Kenya was surveyed and 13 genera containing 51 species with 19 formae were identified. Four new species were recognized, then described as Diplodinium africanum n. sp., Diplodinium namum n. sp., Eudiplodinium kenyensis n. sp., and Ostracodinium iwawoi n. sp. In addition, Buetschlia triciliata and Ostracodinium stokyi were found for the second and third times and described as Hsiungia triciliata n. comb. and Enoploplaston stokyi n. stat., with two formae, respectively. The species composition was similar to that of Indian zebu but several species were considered to originate from African native ruminants. In the percentage composition of genera, the numbers of Entodinium, which normally predominate in rumens in other areas, were very low.     

Acquisition and expression of resistance by Bos indicus and Bos indicus X Bos taurus calves to Amblyomma americanum infestation

Purebred and crossbred Bos indicus calves were infested 1, 2, or 3 times with 10 female and 5 male Amblyomma americanum. Resistance was acquired by both the purebred and the crossbred calves after 1 infestation and resulted in statistically significant decreases in the percentages of females that engorged, the mean weights of engorged females, and the mean weights of egg masses. Comparisons between breeds of the percent of female ticks that engorged during the first and second infestations indicate that purebred B. indicus expressed a stronger acquired resistance to A. americanum more readily than did crossbred animals. However, calves of both genetic compositions displayed similar levels of resistance during a third exposure. All tick-exposed and control animals were skin tested with salivary gland extracts of A. americanum, A. cajennense and Dermacentor andersoni. Control, uninfested calves, did not display significant cutaneous reactivity to these extracts. All calves that had been infested had immediate, 30-min, 5-hr and delayed, 24-hr, skin reactions to Amblyomma species antigens. Reactions to D. andersoni salivary antigens in tests of both purebred and crossbred calves with acquired resistance to A. americanum suggest that Amblyomma species salivary gland antigens might have cross reactive moieties with a salivary extract prepared from D. andersoni. Peripheral blood lymphocyte in vitro responsiveness to Amblyomma species antigens was detected in purebred calves after a first, second, and third infestation, indicating the presence of cells of the immune system capable of recognizing and undergoing blast transformation in response to tick salivary components.(ABSTRACT TRUNCATED AT 250 WORDS)