Serological profile of buffalo (Bubalus bubalis) female calves vaccinated with standard Brucella abortus strain 19 vaccine using rose bengal, 2-mercaptoethanol and complement fixation tests

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ultrasonographic monitoring of early pregnancy development in Murrah buffalo heifers (Bubalus bubalis)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ovarian morphometric characterization and in vitro maturation of oocytes obtained from buffalo (Bubalus bubalis) ovaries - partial results

Buffalo ovaries were collected from a slaughterhouse (Frigol, Brazil) and transported to the laboratory in saline solution at 36 degrees C. The ovaries were dissected to realize the evaluations (weight, length, width and height of the ovary; corpus luteum and dominant follicle diameters). The Cumulus-oocyte complexes (COCs) were recovered by aspiration of 2-8 mm follicles. Selected COCs were matured in TCM 199 supplemented with 10% fetal bovine serum, sodium pyruvate, LH, FSH, estradiol and gentamicin. In vitro maturation was carried out at 38.5 degrees C for 22-24 h and 34-36 h. For the evaluation of the nuclear maturation the oocytes were placed in TCM 199 medium added with type v hialuronidase where the granulosa cells were extracted. The denuded oocytes were transferred to 10 mu l of Hoescht 33342 and the chromosomic configuration was evaluated. The oocytes were classified according to meiosis stage in: Germinal Vesicle, Germinal Vesicle Breakdown, Metaphase I, Metaphase II and Degenerated. The means of weight, length, width and height of the ovary were 3.83 g, 2.27 cm, 1.08 cm and 1.56 cm, respectively. The means of corpus luteum and dominant follicle diameters were 1.40 cm and 7.77 mm. The proportion of oocytes that reached metaphase II stage was: 36.68%.

Structural and ultrastructural characterization of buffalo fetus (Bubalus bubalis) ovarian germinative cells

The objective of the present study was to characterize ovogones, primary oocytes and preantral follicles of buffalo fetus in different ages of gestation. For this, 29 fetuses were collected from a slaughterhouse (Frigol, Brazil) and crown-rump lengths were measured to estimate the fetal age (0-3, 4-6, 7-10 months of gestation). The ovaries were removed and ovarian tissue was processed for classic histology and transmission eletron microscopy examination. The structural evaluation demonstrated that in the first period of the gestation (0-3 months) the buffalo fetus showed ovogones (in mitotic division) and in some cases, the primary oocytes surrounded by somatic cells. In the second period (4-6 months), it was verified that the preantral follicles were completely formed. In the last period (70 month to the end of gestation) the ovaries contained a large amount of preantral follicles, and in some fetuses, antral follicles were observed. The ultrastructural analysis of the ovogones, primary oocytes and preantral follicles showed that these cells have few organelles and the quantity of mitochondria, endoplasmatic reticulum and apparatus Golgi complex is increased as the germinative cells passing from one stage to another.

Morphologic and quantitative evaluation of preantral follicles of buffalo (Bubalus bubalis) in different reproductive phases

The objectives of the present study were to estimate the population of the normal and degenerated preantral follicles of buffaloes in different reproductive phases and to classify the different types of degeneration of ovarian preantral follicles. For this, 18 ovaries were divided in three groups: prepubertal, non pregnat and pregnant adult females. The ovaries were collected from a slaughterhouse (Frigol - Brazil) and processed for classic histological examination. The follicular population was estimated according to Cahill et al. (1979) and calculated according to Gougeon & Chainy (1987). Young buffaloes, pregnant and non pregnat adults presented an average number of 15.5, 3.7 and 8.9 preantral follicles with normal morphology and type I (the degeneration was observed just in oocyte) and II (oocyte and granulosa cells were degenerated) degeneration, respectively. This experiment demonstrated that the proportions of primordial, primary and secondary follicles were affected by reproductive phase. A significant difference was observed considering the proportions of preantral follicles with normal morphology and Type I degeneration in all reproductive phases evaluated. The primary follicles presented more percentage of Type I degeneration that the others follicles and the primordial and primary follicles presented more percentage of Type II degeneration in three reproductive phases evaluated.

Ultrasonographic and endocrine aspects of follicle deviation, and acquisition of ovulatory capacity in buffalo (Bubalus bubalis) heifers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Twelve loci from HSA10, HSA11 and HSA20 were comparatively FISH-mapped on river buffalo and sheep chromosomes

Ten type I loci from HSA 10 (IL2RA and VIM), HSA11 (HBB and FSHB) and HSA20 (THBD, AVP/OXT, GNAS1, HCK and TOP1) and two domestic cattle type II loci (CSSM30 and BL42) were FISH mapped to R-banded river buffalo (BBU) and sheep (OAR) chromosomes. IL2RA (HSA 10) maps on BBU 14q13 and OAR13q13, VIM (HSA 10) maps on BBU14q15 and OAR13q15, HBB (HSA11) maps on BBU16q25 and OAR15q23, FSHB (HSA11) maps on BBU16q28 and OAR15q26. THBD (HSA20) maps on BBU 14q15 and OAR13q15 while AVP/OXT. GNAS1, HCK, and TOP I (HSA20) as well as CSSM30 and BL42 map on the same large band of BBU 14q22 and OAR13q22. All loci were mapped on the same homologous chromosomes and chromosome bands of the two species, and these results agree with those earlier reported in cattle homologous chromosomes 15 and 13. respectively, confirming the high degree of both banding and physical map similarities among the bovid species. Indirect comparisons between physical maps achieved on bovid chromosomes and those reported on HSA10, HSA11 and HSA20 were performed. Copyright (C) 2001 S. Karger AG, Basel.

Phenotype characterization of Staphylococcus species strains isolated from buffalo (Bubalus bubalis) milk

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparative RH Maps of the River Buffalo and Bovine Y Chromosomes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A radiation hybrid map of river buffalo (Bubalus bubalis) chromosome 7 and comparative mapping to the cattle and human genomes

A preliminary radiation hybrid (RH) map containing 50 loci on chromosome 7 of the domestic river buffalo Bubalus bubalis (BBU; 2n = 50) was constructed based on a comparative mapping approach. The RH map of BBU7 includes thirty-seven gene markers and thirteen microsatellites. All loci have been previously assigned to Bos taurus (BTA) chromosome BTA6, which is known for its association with several economically important milk production traits in cattle. The map consists of two linkage groups spanning a total length of 627.9 cR(5,000). Comparative analysis of the BBU7 RH 5,000 map with BTA6 in cattle gave new evidence for strong similarity between the two chromosomes over their entire length and exposed minor differences in locus order. Comparison of the BBU7 RH 5,000 map with the Homo sapiens (HSA) genome revealed similarity with a large chromosome segment of HSA4. Comparative analysis of loci in both species revealed more variability than previously known in gene order and several chromosome rearrangements including centromere relocation. The data obtained in our study define the evolutionarily conserved segment on BBU7 and HSA4 to be between 3.5 megabases (Mb) and 115.8 Mb in the HSA4 (genome build 36) DNA sequence. Copyright (c) 2008 S. Karger AG, Basel.

A radiation hybrid map of river buffalo (Bubalus bubalis) chromosome 1 (BBU1)

The largest chromosome in the river buffalo karyotype, BBU1, is a submetacentric chromosome with reported homology between BBU1q and bovine chromosome 1 and between BBU1p and BTA27. We present the first radiation hybrid map of this chromosome containing 69 cattle derived markers including 48 coding genes, 17 microsatellites and four ESTs distributed in two linkage groups spanning a total length of 1330.1 cR(5000). The RH map was constructed based on analysis of a recently developed river buffalo-hamster whole genome radiation hybrid (BBURH5000) panel. The retention frequency of individual markers across the panel ranged from 17.8 to 52.2%. With few exceptions, the order of markers within linkage groups is identical to the order established for corresponding cattle RH maps. The BBU1 map provides a starting point for comparison of gene order rearrangements between river buffalo chromosome 1 and its bovine homologs. Copyright (C) 2007 S. Karger AG, Basel.

Microsatellite loci isolation from river buffalo using enriched partial genomic libraries

The extensive use of buffalo in agriculture, especially in developing countries, begs for genetic resources to evaluate and improve traits important to local and regional economies. Brazil presents the largest water buffalo populations in the New World, with 1 1 million heads including swamp and river types. To design rational breeding strategies for optimum utilization and conservation of available genetic variability in the Brazilian buffalo's population, it is essential to understand their genetic architecture and relationship among various breeds. This depends, in part, on the knowledge of their genetic structure based on molecular markers like microsatellites. In the present study, we developed six enriched partial genomic libraries for river buffalo using selective hybridization methods. Genomic DNA was hybridized with six different arrays of repeat motif, 5' biotinylated - (CA)(15), (CT)(15), (AGG)(8), (GAAA)(8), (GATA)(8), (AAAAC)(8) - and bound to streptavidin coated beads. The cloning process generated a total of 1920 recombinant clones. Up to date, 487 were directly sequenced for the presence of repeats, from which 13 have been positive for presence of repeats as follows: 9 for di-nucleotide repeats, 3 for tri-nucleotide repeats and 1 for tetra-nucleotide repeat. PCR primer pairs for the isolated microsatellites are under construction to determine optimum annealing temperature. These microsatellites will be useful for studies involving phylogenetic relationships, genome mapping and genetic diversity analysis within buffalo populations worldwide.

First radiation hybrid map of the river buffalo X chromosome (BBUX) and comparison with BTAX

We report the first radiation hybrid map of the river buffalo X chromosome generated from a recently constructed river buffalo (Bubalus bubalis) whole-genome radiation hybrid panel (BBURH5000). This map contains a total of 33 cattle-derived markers, including 10 genes, four ESTs and 19 microsatellites. The markers are distributed in two linkage groups: LG1 contains eight markers spanning 125.6 cR, and LG2 contains 25 markers spanning 366.3 cR. LG1 contains six markers in common with bovine sequence assembly BUILD 3.1. With the exception of BMS2152, the order of these markers on our BBUX map is shuffled when compared to the cow X chromosome (Bos taurus; BTAX). From LG2, two markers (AMELX and BL22) map to a more distal portion of BTAX compared to BBUX. In addition, two pairs of LG2 markers exhibit inversions compared to BTAX (ILSTS017 and ATRX; XBM38 and PPEF1). Alternatively, when compared to the most recent bovine RH map (Bov-Gen 3000rads), BL1098 and BMS2227 from LG1 as well as PLS3 and BMS1820 from LG2 showed inverted positions on the BBUX map. These discrepancies in buffalo and cattle maps may reflect evolutionary divergence of the chromosomes or mapping errors in one of the two species. Although the set of mapped markers does not cover the entire X chromosome, this map is a starting point for the construction of a high-resolution map, which is necessary for characterization of small rearrangements that might have occurred between the Bubalus bubalis and Bos taurus X chromosomes.