Blood group and biochemical polymorphism studies in Bos bubalus

Cattle reagents were used to blood type 158 water buffalo originating from Central Romania.
Out of 36 cattle blood group factors which could be identified with the available reagents only 13 were present in the buffalo.
Three genotypes at the Tf locus in buffaloes were identified by starch gel electrophoresis: TfBB, Tf BC and TfCC. At the Hb locus, only one phenotype Hb A was identified in all samples showing two bands a fast and a slow one.
These data indicate a certain homology between cattle and water buffalo.     

In vivo pelvimetry in buffaloes (Bos bubalus)

The comparison of external and internal, pelvimetry measurement was performed on 79, buffalo heifers, and a high, positive correlation (r=0.90) between the two methods was revealed. The pelvic area of the buffalo heifers increased progressively with the advancing age and with the increase in body weight. The convenience with which external pelvimetry could be performed, the simplicity of the equipment which was used and the accuracy of the method would certainly favor the application of external pelvimetry for routine use.     

Water buffalo (Bubalus bubalus Arnee) allotypes: Identification of two specificities controlled by independent genes

Two water buffalo allotypes (Bl and CI) are described, which are located on distinct low molecular weight molecules. Bl is common to water buffalo and cattle. These two markers are inherited in a simple Mendelian manner and controlled by two independent genes.     

Immune regulation of ovarian function in buffaloes (Bubalus bubalus)

We studied the infiltration of different subsets of immune system cells in the ovarian parenchyma of Egyptian buffaloes during follicular and luteal phases of the estrous cycle. All subsets of leukocytes infiltrated significantly more into corpora lutea (CL) than into Graafian follicles (GF) (P < 0.01) except for plasma cells that were abundant in the GF but not observed in the CL. The number of macrophages, lymphocytes, neutrophils and eosinophils were significantly greater in mature CL than in corpora hemorrhagica (CH) or regressing CL. Moreover, the regressing CL showed significantly more macrophages, lymphocytes and neutrophils than the CH. Large antral follicles were infiltrated with larger number of leukocytes than growing preantral atretic follicles. Macrophages and neutrophils observed in large antral follicles were significantly more abundant in the theca externa than the theca interna (P < 0.01). Only plasma cells were significantly greater in number in the theca intema (P < 0.01). Leukocytes infiltrated significantly more into large mature follicles than large, growing, preantral atretic follicles (P < 0.01). Results of this study reveal the calling of leukocytes in a significant numbers inside the ovarian tissue of buffaloes around the time of ovulation and at luteolysis. It is possible that leukocytes with their powerful bioactive cytokines (IL-1, TNFalpha, GM-CSF, and INF-gamma) may assist in ovarian functions such as ovulation and luteolysis.     

Hemoglobin polymorphism in Italian water buffalo (Bubalus bubalus (Arnee))

Electrophoretic analysis of 794 individual samples of water buffalo hemoglobin shows the presence of three phenotypes, AA, AB and BB. Phenotype AA has only the fast hemoglobin (Hb-fast). Phenotype AB and BB have the fast as well as the slow hemoglobin (Hb-slow). The quantitative ratio Hb-fast: Hb-slow is approximately 68:32 in BB and 84:16 in AB. Several genetic mechanisms are discussed in the light of the biochemical and family data provided.     

Histology of hemal nodes of the water buffalo (Bos bubalus)

Hemal nodes are independent lymphoid organs found in various mammals but are ignored by most immunologists. They seem to play a role in defense against blood-borne infections in some species. The structure of the hemal node has been described in various species but, so far, not in the water buffalo. Specimens were obtained from ten clinically healthy male animals (five calves: 2–3 months old; five bulls: 2–8 years old). Six hemal nodes were obtained from each animal from the mesenteric and perirectal region. The samples were studied by light and transmission electron microscopy. The hemal nodes are bean-shaped or spherical, with one hilus through which the hilus arteries and nerves enter the node and from which veins and lymphatics leave it. The buffalo hemal node has a thin capsule of connective tissue and a few smooth muscle cells. Trabeculae extend from the capsule partially dividing the parenchyma. Subcapsular and trabecular blood sinuses are present. The parenchyma is composed of irregular lymphoid cords rich in erythrocytes, macrophages, and plasma cells and is separated by blood sinuses of variable size engorged with blood. These blood sinuses drain into the trabecular sinuses and then into the subcapsular sinus. In calves, the size of the lymphoid cords is larger than that in adult bulls. Buffalo hemal nodes can be classified as typical hemal nodes, because they are definitely different from hemolymph nodes in other species. They may play a role in filtering the blood.     

基于线粒体12S rRNA基因鉴别混合牛肉及制品的牛种来源

线粒体基因组具有种内高度保守性。它的12S rRNA基因同时具有抗腐蚀、耐高温的特点, 而被用于饲料、鲜肉及肉制品的来源追踪和物种成分鉴别等研究。利用牦牛、普通牛、水牛作为研究材料, 在牛线粒体12S rRNA基因通用引物扩增片段区域发现3种特异的酶切位点, 可用于混合鲜牛肉及制品的牛种来源的鉴别。结果显示: 牦牛12S rRNA扩增片段被酶切为203 bp和250 bp, 普通牛为134 bp和318 bp, 水牛为86 bp和367 bp, 通过测序验证了特异性位点和酶切的正确性; 对样品经过不同温度(100℃、120℃、140℃、160℃和180℃)处理均能扩增到目的条带, 但条带从120℃开始变淡。该方法在鲜牛肉及制品的牛种来源鉴别上具有简单、快速、廉价等优点。     

Fatal intestinal coccidiosis in a three week old buffalo calf (Bubalus bubalus)

The water buffalo (Bubalus bubalus) is important to the economy of several countries, especially in Asia and Brazil. Little is known of the impact of coccidiosis in buffaloes. Cattle and buffaloes are considered to have common species of Eimeria but critical cross transmissions have not been made because it is difficult to raise these hosts coccidian free. Clinical coccidiosis was confirmed post mortem in a 22-day old buffalo calf that died after a 3-4 day illness. Oocysts morphologically identical to Eimeria bareillyi were found in the feces and in sections of small intestine. Oocysts were often pyriform, sometimes with asymmetrical sides. The shorter end was flattened and approximately 5-6 microm wide. Unsporulated oocysts in feces were 23.2-29.5 x 16.5-22 microm in size with an average of 27.2 x 19.3 microm . Schizonts, gamonts, and oocysts were identified in sections of small intestine and they were located in entrocytes of jejunum and ileum. No coccidian stages were seen in sections of colon. This is one of the first confirmed cases of clinical coccidiosis in water buffalo.     

Water buffalo (Bubalus bubalus Arnee) allotypes: Identification of a multiple allelic system

This paper describes two allotypes of water buffaIo controlled by two codominant allelic genes. The third allele is a null allele and behaves as a recessive one. The two detectable serum antigens are termed A1 and A2 aild the third one (as yet undetectable) A0. The A1 antigen was recovered in the third peak and A2 antigen in the first peak following gel filtration through Sephadex G-200. The A1 antigen is common to water buffalo and cattle; the frequency of the corresponding gene (A¹) was the same in both species.     

Detection of genome specific monomorphic loci in Bos taurus and Bubalus bubalis with oligodeoxyribonucleotide probe

A synthetic oligodeoxyribonucleotide probe (OAT36) comprising nine repeats of 5'GACA 3′ and several enzymes were used to analyse cow, (Bos taurus) and buffalo (Bubalus bubalis) genomes and a number of monomorphic loci were detected in both the species. Different animals from the same species showed an almost ‘similar’ monomorphic hybridization pattern but animals from two separate species showed a different ‘genome specific’ pattern. The overall hybridization with any enzyme and probe combination was found to be unique to one species. This forms the basis of genome specific hybridization which is substantiated by our zoo-blot hybridization studies. The evolutionary aspect of these loci in the context of sequence polymorphisms is discussed.     

Effects of beta-carotene, selenium and vitamin A on in vitro polymorphonuclear leukocytic activity in peripartal buffalo (Bubalus bubalus)

The effect of different concentrations of three antioxidans on phagocytic and kill activities of blood polymorphonuclear leukocytes (PMN) isolated from buffaloes during the peripartum period (4 weeks before to 7 weeks after parturition) was investigated in this study. Two concentrations of beta-carotene and vitamin A (10(-6) and 10(-5) M) and one concentration of Se (10(-9) M) were used. Phagocytic activity of PMN treated with beta-carotene (10(-6)M) significantly enhanced (P < 0.05) after parturition (Week 0 until Week 3), whereas the kill activity of the same cells significantly (P < 0.05) increased before and after parturition (at Weeks -4, -3, -2, 0, 1, 2 and 3). The concentration of beta-carotene (10(-5) M) enhanced phagocytosis of PMN only at Weeks 0 and 1 and kill activity at Weeks -4, -3, -2, 0, and 1. Selenium (10(-9)M) significantly (P < 0.05) enhanced phagocytic activity of PMN starting from parturition (Week 0) until Week 3 postpartum. Kill activity increased significantly both before (Weeks -4, -3 and -2) and after (Weeks 0, 1, 2, 3 and 4) parturition. Vitamin A (10(-6) M) significantly enhanced phagocytic activity of PMN at Weeks 0, 1, and 2, whereas, the concentration of beta-carotene (10(-5) M) increased phagocytic activity only at Week 0. Kill activity of PMN increased significantly (P < 0.05) at Weeks -1 and 0 (10(-6)M). These results demonstrate that beta-carotene and selenium significantly enhanced phagocytic and kill activities of PMN isolated from buffaloes around parturition in vitro. Vitamin A enhanced phagocytosis and kill activities but not to the same extent as beta-carotene and selenium. Apparently, the in vitro killing activity of PMN is a distinctive function from phagocytosis and both activities may be enhanced by the use of essential nutrients, especially during the peripartum period. Moreover, beta-carotene is more effective as an antioxidant than vitamin A in enhancing the activities of phagocytic cells.     

Isolation of immunologically active uterine luminal proteins associated with follicular and luteal phases of the ovary in buffalo (Bubalus bubalus)

Uterine luminal proteins (ULP) collected from the genital tract of buffalo during the follicular (Group F) and luteal (Group L) phases of the estrous cycle were chromatographed using sephacryl S-200 gel. Five peaks were detected in each group. Different protein concentrations (10 to 200 microg) from Peaks I and V in each group were examined for immunological activity on polymorph nuclear leukocytic cells (PMNL) in vitro. All concentrations except 10 microg of ULP Peak I (< or = 250 kDa) in Group F enhanced phagocytic activity of PMNL. Peak V (56 kDa) in the same group enhanced phagocytic activity of PMNL only at low protein concentrations (10, 20 and 40 microg protein), while at greater concentrations (80, 150 and 200 microg protein) PMNL activity was suppressed. On the other hand, all protein concentrations from Peak 1 (> or = 250 kDa) in Group L suppressed PMNL activity in a dose-dependent manner. Proteins from Peak V (31 kDa) in Group L suppressed PMNL phagocytic activity at all concentrations but not to the same extent as in Peak I. Electrophoretic analysis of Peaks I and V in both groups revealed only 3 detectable protein bands (subunits) in Peak I and 1 detectable subunit in Peak V. Several additional proteins were probably not detected. The molecular weights of the detected subunits in Peaks I and V in Group F were greater than those in Group L as indicated by SDS-PAGE analysis. The results of this study show that ULP collected from buffalo possessed proteins that modulated phagocytic activity of PMNL in vitro. Proteins collected during the follicular phase, especially Peak I, enhanced phagocytic activity of the PMNL, whereas those collected during the luteal phase (Peaks I and V) suppressed activity. Changes in the molecular weights of ULP detected in this experiment may be related to the changes in phagocytic activity of PMNL tested in vitro.     

Entamoeba bubalus, n.sp., from Carabao

SUMMARY. Twelve carabao in the Philippines were found to be lightly infected with intestinal amoebae. Trophic forms (12 microns in diameter) of this protozoan possessed a definite ectoplasm and a homogeneous endoplasm. They were found only in stained preparations. The nucleus was similar to that of the cyst. All unconcentrated fecal smears contained at least a few cysts (8 microns in diameter). In these forms the cytoplasm usually contained a large vacuole and one or more irregular chromatoidal bodies. The nucleus (2.6 microns in diameter) possessed a pronounced, deeply staining, uniform peripheral ring and a large irregular endosome. There was no periendosomal ring. This amoeba is designated as Entamoeba bubalus , n.sp.     

Diurnal changes in concentration of rumen ciliates and in occurrence of dividing forms in water buffalo (Bubalus bubalus) fed once daily.

When buffalo were fed once daily, significant diurnal variations in concentration of rumen ciliates and occurrence of dividing protozoa were found. Differences in proportions of dividing Entodinium- and Diplodinium-type ciliates were also observed. Results obtained suggest that the range of diurnal fluctuations in rumen protozoa concentration may be related to the percentage of dividing cells in populaitons of these organisms.     

Cloning and characterization of novel cathelicidin cDNA sequence of Bubalus bubalis homologous to Bos taurus cathelicidin-4.

Cathelicidin synthesized by bone marrow cells plays an important role in neutralizing invading pathogens. In the present study, the myeloid cathelicidin cDNA from Bubalus bubalis has been cloned and characterized. RNA from bone marrow of buffalo ribs was extracted, reverse transcribed and amplified using specific pair of primer designed from published cathelicidin-4 cDNA sequences of Bos taurus popularly known as indolicidin. An expected amplified product of 517 bp was obtained, which was cloned and sequenced. Comparison of buffalo cathelicidin and indolicidin sequences reveal that the open reading frames (ORF) of both these two congeners consist of 435 nucleotides with 28 divergent nucleotides and the translated proteins of 144 amino acid residues. Fourteen amino acid residues were found to be dissimilar between these two congeners. The molecular mass of buffalo cathelicidin calculated from the deduced amino acid sequence was 16.23 kDa, which is in close proximity of indolicidin. The sequence comparison with known B. taurus cathelicidin congeners again show 70.8-92.9% identity at nucleotides level and 65-88.3% identity at amino acids level. The maximum similarity of buffalo cathelicidin both at nucleotides level (92.9%) and protein level (88.3%) was found to be with indolicidin. Phylogenetic tree analysis at nucleotides and amino acids level indicate that buffalo, cattle, sheep, pig and equine cathelicidin sequences comprise one clade which are distantly related with human, rabbit and murine cathelicidins. It may be reasonably concluded that buffalo possess the ancestral gene of cathelicidin like that of bovine species.     

Diurnal changes in concentration of rumen ciliates and in occurrence of dividing forms in water buffalo (Bubalus bubalus) fed once daily.

When buffalo were fed once daily, significant diurnal variations in concentration of rumen ciliates and occurrence of dividing protozoa were found. Differences in proportions of dividing Entodinium- and Diplodinium-type ciliates were also observed. Results obtained suggest that the range of diurnal fluctuations in rumen protozoa concentration may be related to the percentage of dividing cells in populaitons of these organisms.