Mitochondrial DNA analyses of Indian water buffalo support a distinct genetic origin of river and swamp buffalo

Water buffalo (Bubalus bubalis) is broadly classified into river and swamp categories, but it remains disputed whether these two types were independently domesticated, or if they are the result of a single domestication event. In this study, we sequenced the mitochondrial D-loop region and cytochrome b gene of 217 and 80 buffalo respectively from eight breeds/locations in northern, north-western, central and southern India and compared our results with published Mediterranean and swamp buffalo sequences. Using these data, river and swamp buffalo were distinguished into two distinct clades. Based upon the existing knowledge of cytogenetic, ecological and phenotypic parameters, molecular data and present-day distribution of the river and swamp buffalo, we suggest that these two types were domesticated independently, and that classification of the river and swamp buffalo as two related subspecies is more appropriate.     

Concise synthesis of a buffalo milk pentasaccharide derivative

An efficient synthesis of buffalo milk pentasaccharide derivative via a 3+2 strategy is described. The use of a trisaccharide isopropyl thioglycoside as a latent glycosyl donor and the application of two well-defined regioselective glycosylations significantly simplified the target preparation.     

Isolation and characterization of a sialo-glycopeptide from buffalo colostrum

A sialoglycopeptide was isolated from buffalo colostrum in pure form by chromatography on Sephadex G-25 and QAE-Sephadex A-25. This was found to be homogeneous by cellulose acetate membrane electrophoresis and reverse phase HPLC. It consisted of fucose, galactose, mannose,N-acetyl glucosamine andN-acetyl neuraminic acid in the ratio 12341, and aspartic acid, serine, threonine, proline and glutamic acid were the major amino acids. Glycine was identified as the N-terminal amino acid residue. The structure elucidation of the carbohydrate moiety was carried out by methylation analysis, mass spectrometry,¹H-NMR spectroscopy and the probable structure was revealed to be that of a complex biantennary type.     

Heterogeneity in buffalo pituitary prolactin

The Ellis procedure of serial extraction of gonadotropins and growthhormone (GH) followed by alkaline ethanol extraction was adopted to processfreshly frozen buffalo pituitaries. The procedure after slight modificationwas found very useful as more than 2 mg of GH free immunoreactive prolactin(PRL) could be isolated from each gram of wet pituitary tissue. Further, thebiochemical purity and immunobiological potency of the extracted PRL,designated as P-I, was comparable with that of the highly purified samplesof homologous and heterologous PRLs. No non-PRL protein was detectable inP-I. Micro-heterogeneity with regard to size, charge, co- andpost-translational modifications was also investigated under differentconditions of extraction and at different stages of purification.Immunological and biological potencies were compared in homologouscompetitive enzyme linked immunosorbent assay (ELISA) developed for buffaloPRL and in rat Nb2 lymphoma proliferation assay respectively. Structuralheterogeneity was observed in all the preparations checked including freshpituitary homogenate and highly purified hormone. Nevertheless a 25 Kspecies corresponding to the hormone monomer was always the only paramountform comprising more than 90% of the total PRL protein in all thesamples including P-I. Similar size forms were observed in all preparationsand were found to be equivalents of monomers, dimers, covalent-andnon-covalent multimers, disulphide bridged forms and cleaved fragments.Other sibling species identified were glycosylated PRL, charge isoforms andforms that perhaps differed in their extractability from the pituitarytissue. Strong apparent size heterogeneity was displayed by the monomericbuffalo PRL. In light of these observations and the information on thestructural and functional significance and the consequences of polymericforms, the use of a heterogeneous PRL (P-I) as a reference hormone isrecommended for a valid assay.     

Immunosuppressive activity in buffalo placenta

Immunosuppressive activity in buffalo placenta was evaluated by measuring proliferation of lymphocytes in presence of phytohaemagglutinin (PHA) alone or PHA plus placental proteins. The immunosuppressive activity was dose-dependent over the protein concentration range of 10-50 micrograms/ml. Proteins from both cotyledon and non-cotyledon portions of placenta exhibited immunosuppressive activity. Fractions obtained with 0-40, 40-60 and 60-80% saturated ammonium sulphate exhibited 70, 73 and 75% suppression, respectively. PBS-soluble placental proteins were resolved on G-100 column into three peaks that exhibited 69 (peak 1), 55 (peak 2) and 73% (peak 3) suppression. Exogenously added interleukin-1 (IL-1) failed to reverse the suppression caused by buffalo placental proteins.     

Freezing of swamp buffalo semen

The French mini straw technique (Society I.M.V.) was used to preserve semen of Indonesian swamp buffalo (Bubalus bubalis) in a lactose based extender with and without the removal of seminal plasma. Extended semen with 60–70% motility before freezing showed 60 and 40–50% motility after thawing at 4°C for 5 and 180 min. respectively. Nine of 13 cows conceived to a single insemination with frozen semen. Neither post-thawing motility nor conception was improved by removing seminal plasma before freezing     

Meiotic peculiarities in hybrid buffalo

The two varieties of the Water buffalo (Bubalis bubalis var. bubalis and Bubalis bubalis var. carabao) have 2n = 50 and 2n = 48 karyotypes, respectively. The F1 hybrids are thought to exhibit a karyotype of 2n = 49 and are known to be fertile. Meiosis was studied in 10 hybrid water buffalo bulls. Karyotypes of the bulls were prepared from leukocyte cultures and testicular biopsy samples in a routine manner and examined. Phenotypically the bulls showed characteristics of the hybrid buffalo. Five of the bulls carried 2n = 50 and 5 had 2n = 49. Multivalent chromosomes were found in diakinesis (metaphase I) cells of bulls with 2n = 49 karyotypes. Synapses were found in bulls of both karyotypes.     

Heterogeneity in buffalo lutropin.

Lutropin (LH-1) from water buffaloes has been shown to exhibit microheterogeneity in the N-terminal amino-acid sequence of its alpha-subunit. The beta-subunit did not exhibit such microheterogeneity. Another protocol of purification yielded a preparation of buffalo LH (bu LH-2) different from the buffalo LH-1 in certain physico-chemical properties like ease of dissociation into subunits, sugar composition, isoelectric point, and elution profile on S-200. Data appear to indicate the presence of more than one form of buffalo lutropin.     

Synteny mapping in river buffalo

The cosegregation of ten coding loci has been investigated, in a panel of 37 somatic cell hybrids resulting from the fusion of a hamster cell line and river buffalo lymphocytes, by use of Southern hybridization technique. Five syntenic groups, TCRB-PGY3, ASS-ABL, FUCA1P-CRYG, MBP-YES1, and CGN1-ACTA1, previously assigned to cattle as U13, U16, U17, U28, and U29 respectively, were also found to be syntenic in buffalo. Based on the extensive syntenic conservation and banding homology between cattle and river buffalo, comparative mapping predicts the localization of these syntenic groups on river buffalo Chromosomes (Chrs) :BBU7, BBU12, BBU2q, BBU22, and BBU4q respectively as they have been previously localized on cattle Chrs BTA4, BTA11, BTA2, BTA24 & BTA28. Received: 2 April 1996 / Accepted: 4 July 1996     

Banking North American buffalo semen

The purpose of this study was to develop a procedure to collect and preserve semen from wood bison (Bison bison athabascae) and plains bison (Bison bison bison). Semen samples from three wood and three plains bison bulls were collected by electroejaculation from June through October. In addition, sperm was collected from the cauda epididymis of seven plains bison. Semen was cryopreserved using two commercially available cryopreservation media, an egg yolk-based medium (Triladyl), and a medium free of products of animal origin (Andromed). Sperm morphology and motility were recorded on fresh and post-thawed semen samples. Total sperm motility was not different between plains and wood bison for the months of June (50%), July (69%) and October (54%). However, total sperm motility for wood bison was higher (P < 0.05) than plains bison for the months of August and September (August: 80% vs 55%; September: 73% vs 40%). Plains and wood bison did not differ in mean total and mean progressive motility (35 and 15%, respectively) of frozen-thawed sperm samples. The post-thaw motility of Triladyl-treated sperm was higher (P < 0.05) than Andromed-treated sperm (35% vs 13%, respectively). Interestingly, post-thawed epididymal spermatozoa had higher total motility (P < 0.05) than post-thawed electroejaculated sperm when cryopreserved with a medium free of products of animal origin (Andromed; 35% vs 9%, respectively). In conclusion, we used electroejaculation to collect high quality bison semen, and cryopreserved it for future needs.     

Characterization of a 59 kDa gelatin-binding fragment of buffalo plasma fibronectin

Limited proteolysis of buffalo plasma fibronectin (FN) by thermolysin yielded four gelatin-binding fragments of which, the major 59 kDa fragment, GBF1, was isolated by gelatin-Sepharose and heparin-Sepharose affinity columns. GBF1 appeared during early phase of thermolysin digestion and remained intact even after 4 hr of digestion. GBF1 may be similar to 56 kDa gelatin-binding fragment of FNs from human and hamster plasma. But, it is more resistant to thermolysin cleavage. The fragment binds to heparin with low affinity. On the basis of the structure of human plasma FN, the modular structure of GBF1 may be given as: 6Fn1 1Fn2 2Fn2 7Fn1 8Fn1 9Fn1 1Fn3. Biophysical properties of GBF1 suggest an expanded native conformation. The interaction of the fragment with gelatin is pH-dependent and independent of NaCl concentration.     

Cellulolytic bacteria in buffalo rumen

The influence of three different feeds, wheat straw, sorghum and berseem, on total and cellulolytic bacterial counts in the buffalo rumen at different time intervals from 0 to 8 h after feeding was studied. Berseem feeding supported maximum growth of rumen bacteria in general and cellulolytic bacteria in particular. Wheat straw supported the poorest growth.
The types of cellulolytic bacteria recovered from the rumen of adult buffaloes were Ruminococcus albus, R. flavefaciens, Bacteroides succinogenes, Butyrivibrio fibrisolvens, Clostridium lochheadii, Cl. longisporum and other Clostridium spp. Cellulolytic cocci were present in smaller numbers than rod forms in the rumen of wheat-straw-fed buffaloes, whereas the cocci outnumbered rod forms in sorghum-and berseem-fed buffaloes.     

N-terminal groups of buffalo thyroglobulin

N-Terminal analysis of purified buffalo thyroglobulin by the fluorodinitrobenzene method of Sanger yielded about 1.5 moles of DNP-glutamic acid per mole of buffalo thyroglobulin. No water-soluble DNP-amino acid was detectable as N-terminal. The presence of glutamic acid has been confirmed by Edman degradation and characterization of the PTH-amino acid in different solvent systems, and also after regeneration of free amino acid from PTH-amino acid in butanol-acetic acid-water (4:1:5, v/v) system. This is in contrast to the occurrence of aspartic acid or asparagine as N-terminals for several other mammalian thyroglobulins.