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PubMed1987

[Rotavirus enteritis in buffaloes].

Immunodiffusion was used to investigate a total of 492 serum samples of buffaloes--buffalo calves up to the age of 6 months and buffalo heifers--on both private and public farms. Positive serological reagents for rotaviruses were established in 73.4 per cent of the cases. The study of the same samples via the blocking ELISA test revealed the presence of 86.2 per cent positive cases. It was demonstrated that this type of the ELISA technique was a more precise method for the demonstration of group specific rotavirus antibodies in buffaloes. The results obtained spoke of the widely occurring rotavirus infections among the buffalo populations in this country. Particularly intense was the circulation of rotaviruses in buffalo calves aged up to six months. The study of 46 fecal samples from buffalo calves with diarrhea by means of the immunodiffusion and ELISA tests revealed that in as many as seven of them there were rotaviruses. This unquestionably demonstrated the participation of these agents in the etiology of enteritis in buffaloes.

Animals↗
PubMed1979

Experiments on the Babesia bigemina carrier state in East African buffalo and eland.

Three captive East African buffalo (Syncerus caffer) were injected with 5 x 109 bovine erythrocytes with Babesia bigemina pathogenic for cattle. None of the buffalo showed signs of disease, but Babesia were found in blood smears for four days indicating their multiplication. The carrier state of Babesia bigemina was established in two buffalo for 28 and 147 days respectively by sub-inoculation into splenectomized calves, while no parasites were demonstrated 28 days after injection in the third buffalo. Two buffalo shot in Kenya did not appear to be carriers of Babesia gibemina, as shown by sub-inoculation of blood in splenectomized calves. Two captive eland (Taurotragus oryx) were inoculated and their blood investigated as with the experimental buffalo. Neither eland appeared to harbour Babesia bigemina 28 days after injection, even though one was splenectomized. The experimental data indicate that the East African buffalo can be a carrier of Babesia bigemina for at least 5 months, while this parasite did not survive in eland. Buffalo may play a role in the epidemiology of babesiosis in cattle, while eland most likely do not.

Animals↗
PubMed2002

Culture-derived Babesia orientalis exoantigens used as a vaccine against buffalo babesiosis.

Buffalo babesiosis represents a major problem for the livestock industry in China. The pathogen of this disease was isolated and identified as Babesia orientalis and subsequently propagated in vitro, using the microaerophilus stationary phase culture system. The aim of this study was to determine the efficacy of exoantigens derived from B. orientalis cultures to induce a protective immunity against challenge exposure to virulent organisms in laboratory and field. The results showed that exoantigens of B. orientalis could induce a high degree of protection against challenge with a virulent B. orientalis strain. The animals in the vaccinated group exhibited a slight decrease in haemoglobin levels and blood cell counts, whereas animals in the control group showed typical clinical symptoms and died between days 11 and 16 after challenge. In another approach, 82 buffaloes kept in B. orientalis-endemic areas were injected with exoantigens plus Freund's adjuvant in two doses at an interval of 2 weeks, one month before the endemic period. An additional 86 buffaloes served as unvaccinated controls. During the endemic period, the clinical signs of all buffaloes were monitored and the antibody response was investigated, using the latex agglutination test. During the endemic period, none of the vaccinated buffaloes showed clinical signs or died, whereas five control buffaloes showed clinical signs and two of them died. The mean antibody titre of vaccinated animals was higher than that of the control animals. These results showed that the culture-derived exoantigens of B. orientalis could be used as a vaccine to prevent buffalo babesiosis.

Animals↗
PubMed1994

Characterization of immune response and duration of protection in buffaloes immunized with haemorrhagic septicaemia vaccines.

Two of the three buffaloes immunized with a non-adjuvanted broth bacterin were found to be protected against experimental challenge at 6 weeks but not at 3 months post-challenge. Similarly all buffaloes (4/4) immunized with alum-precipitated vaccine were protected at 6 months but only 1 of the 2 vaccinated animals were protected at 12 months post-immunization. On the other hand, buffaloes immunized with an oil adjuvant and a double emulsion vaccine were completely protected at 12 months post-immunization. Statistically significant differences between immunized versus non-immune animals became evident at 3 months post-immunization, although analysis of cumulative antibody titres of pre-challenge sera of vaccinated buffaloes surviving versus those succumbing to experimental challenge revealed significant by higher antibody titres in the former as compared to the latter group. These results suggested that there was a relationship between ELISA antibody titres and active protection in buffaloes. There also appeared to be a relationship between cutaneous delayed-type hypersensitivity and active protection in buffaloes. Preliminary analysis of the antibody isotype distribution in the pre-challenge sera of 2 buffaloes vaccinated with the oil adjuvant vaccine revealed predominance of IgG1 and IgG2 subclasses whose role in protection against haemorrhagic septicaemia was not eludicated.

Animals↗
PubMed2005

Isolation and characterization of heparin and gelatin binding buffalo seminal plasma proteins and their effect on cauda epididymal spermatozoa.

Seventy semen ejaculates were obtained from 14 Murrah buffalo bulls and were subjected to plasma separation immediately after collection by centrifugation at 2000 rpm for 20 min and stored in liquid nitrogen until analysis. In the seminal plasma the total protein concentration were estimated and the heparin and gelatin binding (HB and GB) proteins were isolated using heparin and gelatin affinity column chromatography. The molecular weight of individual isolated HB and GB protein was determined by SDS-PAGE analysis. Buffalo bull spermatozoa was collected from cauda epididymis under aseptic conditions and was used for the in vitro fertility tests (i.e. bovine cervical mucus penetration test (BCMPT) and hypo-osmotic swelling test (HOST)). The heparin and gelatin binding buffalo seminal plasma proteins were used in six concentrations i.e. 10, 20, 30, 40, 50 and 60 microg/ml to test their effect on in vitro fertility assessment of cauda epididymal spermatozoa. The overall mean values of total protein, HB and GB proteins were recorded as 29+/-2.7, 2.61 and 0.2mg/ml, respectively. Eighteen total protein bands were observed in the range of 12-127 kDa. Eight major HB proteins were isolated in the range of 13-71 kDa. Seven major GB proteins were isolated in the range of 13-61 kDa in the buffalo seminal plasma. The mean penetration distance (mm) travelled by the buffalo cauda spermatozoa was maximum in HB proteins (26.9+/-0.6) followed by GB proteins (25.4+/-0.6) and control (21.2+/-1.4). The difference in BCMPT values between protein treated and control group was significant (P<0.05). Almost similar trend in the effect of protein on values of HOST percentage in both HB and GB proteins treated semen samples were recorded (66.4+/-0.65 and 66.1+/-0.6, respectively). The difference in HOST values between proteins treated and control group (50.4+/-2.0) was significant (P<0.05). The present results indicate that among the isolated proteins, 4 proteins were commonly seen in both the heparin and gelatin-sepharose affinity column chromatography, and the addition of buffalo seminal plasma proteins improved the in vitro sperm functions (40 microg/ml gave best results) of buffalo cauda spermatozoa.

Animals↗
PubMed2005

Development of water buffalo (Bubalus bubalis) embryos from in vitro matured oocytes reconstructed with fetal skin fibroblast cells as donor nuclei.

The present study was carried out to explore the feasibility of using buffalo fetal skin fibroblasts as donor nuclei and to find out the developmental competence of embryos following transfer of these nuclei to in vitro matured enucleated buffalo oocytes. Skin cells were isolated from 1 to 2-month-old fetuses obtained from slaughterhouse, by enzymatic digestion (0.5% w/v trypsin +0.05% w/v collagenase in Dulbecco's PBS) for 15-20 min. The cells were washed 4 times with Dulbecco's PBS and then once with RPMI-1640+10% FBS by centrifugation at 600 x g. The cells were then cultured in the same medium in a CO2 incubator (5% CO2 in air) at 38.5 degrees C for 2-3 days. Cumulus-oocyte complexes (COCs) collected from slaughterhouse buffalo ovaries were subjected to IVM in the IVM medium (TCM-199 + 5 microg/ml FSH-P + 10 microg/ml LH+10% FBS) for 20-22 h in a CO2 incubator (5% CO2 in air) at 38.5 degrees C. Oocytes were denuded with 0.1% trypsin followed by repeated pipetting and then enucleated by aspirating the first polar body with 10-15% of nearby cytoplasm with a micromanipulator. Two different types of donor cells (growing cells and those arrested with cytochalasin-B) were used for reconstruction of oocytes. The reconstructs were electro fused and incubated in the activation medium (TCM-199 + 8 microg/ml cytochalasin-B+10% FBS) for 4 h. These were then cultured in IVC medium (TCM-199+10% FBS) in a CO2 incubator (5% CO2 in air) at 38.5 degrees C for 48 h. The cleaved embryos were then co-cultured with buffalo oviduct cells in embryo development media (EDM). Out of 119 denuded matured oocytes which were enucleated and reconstructed with growing cells, 78 (65.5%) were electro fused, activated and cultured, out of which 4 (5.1%) reconstructs cleaved and developed to 2-cell stage, 3 (3.8%) reached to 4-cell stage and 3 (3.8%) reached to 8-cell stage. In the synchronized group, out of 62 denuded matured oocytes which were reconstructed with cytochalasin-B blocked cells, 40 (65%) were electrofused, activated and cultured, out of which 4 (10%) developed to 2-cell stage, 3 (7.50%) to 4-cell stage, 2 (5.0%) to early morula stage and 1 (2.50%) to blastocysts stage. These results suggest that buffalo fetal skin fibroblasts could be used as donor nuclei for the production of buffalo embryos after nuclear transfer to enucleated in vitro matured buffalo oocytes.

Animals↗
PubMed2005

Conception rate after fixed time insemination following ovsynch protocol with and without progesterone supplementation in cyclic and non-cyclic Mediterranean Italian buffaloes (Bubalus bubalis).

The aim of this study was to test the effect of progesterone supplementation to Ovsynch protocol in cyclic and non-cyclic Mediterranean Italian buffaloes on conception rate after fixed time artificial insemination. From 169 pluriparous buffaloes, 2 groups were identified and subjected to: (1) Ovsynch protocol (OV; n=83) and (2) Ovsynch protocol with the supplementation of progesterone from days 0 to 7 (OV+PROG.; n=86). All cows were inseminated 16-20 h after the second GnRH administration. Within each group, non-cyclic buffaloes were identified (OV=21 and OV+PROG.=20). Overall conception rate was significantly higher in cyclic compared to non-cyclic buffaloes: 43.7% versus 17.0%, respectively, P=0.001. A significant effect of progesterone supplementation on conception rate was observed in non-cyclic buffaloes (30% versus 4.7%, P=0.04) but not in cyclic buffaloes (51.5% versus 35.7%, P=0.077). Collectively, the presence of a large follicle (>or=10 mm) detected at the beginning of the Ovsynch protocol by ultrasound significantly affected conception rate (44% versus 8%, P=0.01). The findings of the present study suggest that (i) progesterone supplementation to the Ovsynch protocol in buffaloes increases conception rate in non-cyclic animals, (ii) the presence of a large follicle at the beginning of the Ovsynch protocol is a determining factor for a successful synchronization of ovulation and high conception rates and (iii) ultrasound monitoring can improve the overall efficiency by selectively identifying more suitable cycling animals carrying a responsive follicle at the time of first GnRH administration.

Animals↗
PubMed2005

Efficacy of the Ovsynch protocol for synchronization of ovulation and fixed-time artificial insemination in Murrah buffaloes (Bubalus bubalis).

Two experiments were conducted to assess the timing and synchrony of ovulation, plasma LH concentrations, and pregnancy rate in Murrah buffaloes (Bubalus bubalis) treated with the Ovsynch (GnRH-PGF(2 alpha)-GnRH) protocol. In Experiment 1, 10 non-lactating cycling buffaloes received 10 microg of a GnRH analogue i.m. (buserelin acetate) without regard to the stage of the estrous cycle (day of treatment, day 0), followed by 25mg of PGF(2 alpha) i.m. (dinoprost thromethamine) 7 days later. A second-treatment of the same GnRH analogue (10 microg, i.m.) was given 48 h after PGF(2 alpha). Ovulation was confirmed by transrectal palpation (at 2-h intervals) from the second-GnRH treatment to detection of ovulation or up to 96 h after the second-GnRH treatment. Plasma LH concentrations were determined in blood samples collected at 15-min intervals for 6h, starting at the second-GnRH treatment, and thereafter at 2-h intervals until 2h after detection of ovulation. Ovulation occurred in 9/10 buffalo (90%) 23.3+/-1.3h (mean+/-S.E.M.; range 20--32 h) after the second-GnRH treatment. Peak LH concentrations 13.5+/-3.5 ng/mL (range 3.9--40.0 ng/mL) occurred 2.1+/-0.1h (range 1.2-3.0 h) after the second-GnRH treatment. In Experiment 2, 15 lactating, cycling buffaloes were subjected to the Ovsynch protocol, with fixed-time AI 12 and 24h after the second-GnRH treatment and 75 lactating buffaloes were inseminated, approximately 12h after detection of spontaneous estrus. Pregnancy rates were 33.3% for TAI and were 30.7% for buffaloes inseminated following spontaneous estrus (P=0.84). In conclusion, the Ovsynch protocol effectively synchronized ovulation in Murrah buffaloes and resulted in conception rates (to two fixed-time inseminations) that were comparable to those achieved with a single AI after detection of spontaneous estrus.

Animals↗
PubMed2006

Rapid sexing of water buffalo (Bubalus bubalis) embryos using loop-mediated isothermal amplification.

Loop-mediated isothermal amplification (LAMP) is a novel DNA amplification method that amplifies a target sequence specifically under isothermal conditions. The objective of this study was to identify a Y chromosome-specific sequence in water buffalo and to establish an efficient procedure for embryo sexing by LAMP. The homologues of a Y chromosome-specific sequence, bovine repeat Y-associated.2, in swamp and river buffalo were cloned, and designated swamp buffalo repeat Y-associated.2 and river buffalo repeat Y-associated.2, respectively. Sexing by LAMP was performed using primers for swamp buffalo repeat Y-associated.2. A 12S rRNA was also amplified by LAMP as a control reaction in both male and female. The minimal amount of the template DNA required for LAMP appeared to be 0.1-10 pg. The sensitivity was further examined using swamp buffalo fibroblasts as templates. When fibroblasts were lysed with NaOH, the minimal cell number required for detection of both male-specific and male-female common DNA appeared to be two cells, whereas correct determination of sex could not be achieved using fibroblasts lysed by heat denaturation. Embryo sexing was also performed using blastomeres from interspecies nuclear transfer embryos. The sex determined by LAMP for blastomeres corresponded with the sex of nuclear donor cells in analyses using four or five blastomeres as templates. The LAMP reaction required only about 45 min, and the total time for embryo sexing, including DNA extraction, was about 1 h. In conclusion, the present procedure without thermal cycling and electrophoresis was reliable and applicable for water buffalo embryos.

Animals↗
PubMed2005

Distribution of G (VP7) and P (VP4) genotypes in buffalo group A rotaviruses isolated in Southern Italy.

Group A rotaviruses are established agents of disease in buffalo calves. Early epidemiological studies in Italian buffalo herds revealed the predominance of strains with G8 specificity and detected strains with the rare, RRV-like, VP4 P[3] genotype. To acquire additional information on the VP4 and VP7 specificities of buffalo rotaviruses, a total of 125 fecal samples were collected from buffalo calves affected with diarrhoea, in seven dairy farms in Southern Italy. Rotaviruses were detected in 21 samples (16.8%) by an immunochromatographic assay and by reverse transcription-PCR (RT-PCR). Analysis of the VP7 gene revealed that 57% (12 of 21) of the isolates were G6, 23.8% were G8 (5 of 21) and 19% (4 of 21) were G10. Analysis of the VP4 revealed that 71.4% (15 of 21) of the isolates were P[5] and that 28.6% (6 of 21) were P[1]. The most common combination of G and P types was P[5],G6 (57%), followed by P[1],G10 (19%), P[5],G8 (14%) and P[1],G8 (9.5%). While P[5],G6 rotaviruses are very common in Italian bovine herds, the antigenic combination P[1],G10 is unusual and presumably derives from reassortment between P[1] and G10 strains, that appear to be more frequent in buffaloes and bovines, respectively. The presence of bovine-like G and P serotypes suggests that in Italy the epidemiology of buffalo rotaviruses overlaps the epidemiology of bovine rotaviruses, presumably because of the strict species affinity and/or of the intermingled distribution over the same geographical areas of the buffalo and bovine herds.

Animals↗
PubMed2005

Antibodies against bovine herpesvirus 4 are highly prevalent in wild African buffaloes throughout eastern and southern Africa.

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world. Interestingly, a survey of wild African buffaloes mainly from the Maasai Mara Game Reserve in Kenya revealed that 94% of the animals tested had anti-BoHV-4 antibodies [Rossiter, P.B., Gumm, I.D., Stagg, D.A., Conrad, P.A., Mukolwe, S., Davies, F.G., White, H., 1989. Isolation of bovine herpesvirus-3 from African buffaloes (Syncerus caffer). Res. Vet. Sci. 46, 337-343]. These authors also proposed that the serological antigenic relationship existing between BoHV-4 and alcelaphine herpesvirus 1 (AlHV-1) could confer to BoHV-4 infected buffaloes a protective immune response against lethal AlHV-1 infection. In the present study, we addressed two questions related to Rossiter et al. paper. Firstly, to investigate the role of the African buffalo as a natural host species of BoHV-4, the seroprevalence of anti-BoHV-4 antibodies was analysed in wild African buffaloes throughout eastern and southern Africa. A total of 400 sera was analysed using two complementary immunofluorescent assays. These analyses revealed that independently of their geographical origin, wild African buffaloes exhibit a seroprevalence of anti-BoHV-4 antibodies higher than 68%. This result is by far above the seroprevalence generally observed in cattle. Our data are discussed in the light of our recent phylogenetic study demonstrating that the BoHV-4 Bo17 gene has been acquired from a recent ancestor of the African buffalo. Secondly, we investigated the humoral antigenic relationship existing between BoHV-4 and AlHV-1. Our results demonstrate that among the antigens expressed in AlHV-1 infected cells, epitope(s) recognised by anti-BoHV-4 antibodies are exclusively nuclear, suggesting that the putative property of BoHV-4 to confer an immune protection against AlHV-1 relies on a cellular rather than on a humoral immune response.

Animals↗
PubMed2003

Buffalo and cattle hybrid embryo development is decreased by caffeine treatment during in vitro fertilization.

Water buffalo are renowned for difficulties in the implementation of assisted reproductive technologies, with both males and females being problematic. In this study, we used cattle oocytes to assess the effect of treatments with heparin and caffeine on buffalo spermatozoa and subsequent fertilization and embryo development in vitro. There was no significant difference between buffalo and bovine spermatozoa in the events associated with fertilization. Fertilization of cattle oocytes with buffalo spermatozoa resulted in 7.8% of oocytes developing into hybrid embryos. A difference in the developmental capability of hybrid embryos compared with the cattle control was observed. This has not been previously reported. The subsequent transfer of a limited number of hybrid embryos did not produce a viable pregnancy. However, control treatments in this experiment also failed to achieve pregnancy, so objective data is not available to provide conclusions about the developmental competence of the buffalo and cattle hybrid embryos. Optimal spermatozoa capacitation treatments achieved 61% fertilization and 21% zygote cleavage into two cell embryos. There was no significant difference in fertilization or development due to heparin or spermatozoa concentrations. However, treatment of buffalo and cattle spermatozoa with caffeine significantly decreased embryo cleavage but also tended to decrease embryo development to the blastocyst stage. These studies suggest that problems with reproduction in buffalo may reside with biological mechanisms associated with the oocyte that are often complicated by poor male reproductive performance. Selection for bull fertility would prevent some of these complications.

Animals↗
PubMed2003

Bovine and buffalo in vitro embryo production using oocytes derived from abattoir ovaries or collected by transvaginal follicle aspiration.

This study was undertaken in order to evaluate the effect of oocyte source (live animals and abattoir ovaries) on subsequent embryo development in buffalo (Bubalus bubalis). Cow ovaries were also collected as oocyte donors for in vitro embryo production (IVEP). Three hundred thirty-eight oocytes were recovered by ovum pick up (OPU, Group A) from 8 pluriparous buffalo cows, while 1127 and 1457 oocytes were aspirated, respectively, from buffalo (Group B) and bovine (Group C) slaughterhouse ovaries. Cumulus enclosed oocytes (COCs) suitable for IVEP were in vitro matured (IVM), fertilized (IVF) and cultured (IVC) to the tight morula (Tm) and blastocyst (Bl) stage. Within buffalo species Group A had a higher Bl yield (29.7 % versus 19.9%; P<0.05) and a lower proportion of embryos arrested at Tm stage (11.1% versus 22.3%; P<0.05) than Group B. Within slaughterhouse groups cattle oocytes had a higher cleavage rate (83.9% versus 64.8%; P<0.05) and yielded 49.2% more blastocysts than buffalo. However, when data are related to the total number of cleaved oocytes, only 13.7% more blastocysts were produced in cattle than in buffalo.In conclusion, in buffalo species the source of oocytes significantly affected post-fertilization embryo development, as demonstrated by the higher Bl yields derived from OPU-derived oocytes.A higher overall IVEP efficiency, mainly related to the higher cleavage rate, was recorded in cattle compared with buffalo when ovaries from an abattoir were used as oocyte donors.

Abattoirs↗
PubMed2003

Brazilian isolates of Trypanosoma (Megatrypanum) theileri: diagnosis and differentiation of isolates from cattle and water buffalo based on biological characteristics and randomly amplified DNA sequences.

We detected and cultivated isolates of Trypanosoma (Megatrypanum) theileri from cattle and water buffaloes in São Paulo state, southeastern Brazil, which were characterized by comparing morphological, growth and molecular features. Although isolates from cattle and water buffalo were morphologically indistinguishable, they differed in their growth characteristics. A dendrogram based on randomly amplified polymorphic DNA (RAPD) patterns indicated close-genetic relationships among all isolates from both species, which were all tightly clustered together and distant from Megatrypanum species from wild mammals. In addition, isolates within the T. theileri-cluster were clearly segregated into two host-associated groups. The sequence of a synapomorphic RAPD-derived DNA fragment (Tth625), which was shared by all T. theileri trypanosomes from cattle and buffalo but not detected in any of 13 other trypanosome species, was used as target for a conventional T. theileri-specific PCR assay. We also defined RAPD fragments (Tthc606 and Tthb606) that distinguished cattle from buffalo isolates. Thus, distinct growth features and genetic variability distinguished between isolates from cattle and water buffaloes of the same geographic origin, suggesting an association of these isolates with their host species. The trypanosomes from water buffalo reported here are the first T. theileri-like isolates from the Asiatic buffalo (Bubalus bubalis) to be continuously cultured and compared with cattle isolates using biological and molecular methods.

Animals↗
PubMed2003

Validation of a sensitive enzymeimmunoassay for 13,14-dihydro-15-keto-PGF2alpha in buffalo plasma and its application for reproductive health status monitoring.

A simple, sensitive and direct enzymeimmunoassay (EIA) procedure on microtitre plates using the second antibody coating technique was standardized and validated for the determination of 13,14-dihydro-15-keto PGF2alpha (PGFM) in unextracted buffalo plasma. The assay was carried out directly in 20 microl of buffalo plasma. PGFM standards prepared in charcoal stripped hormone-free plasma were used. The sensitivity of the assay was 0.4 pg/well, which corresponded to 20 pg/ml plasma. Plasma volumes for the assay ranging from 10 to 50 microl did not influence the PGFM standard curve; however, a slight drop in the OD450 was observed with higher plasma volumes. Biological validation of the assay was carried out in buffalo plasma samples obtained during physiological states of cyclicity, peri-estrus, post-insemination, reproductive tract infection and persistent corpus luteum conditions. A pulsatile pattern of plasma PGFM release was observed prior to estrus when PGFM was determined in blood samples collected at hourly intervals of time. The PGFM pulsatility was not observed when blood sampling frequency of either 4 or 12 h was considered. The PGFM levels stayed high in peripheral circulation of buffaloes with reproductive tract infections and remained low throughout the sampling period in buffaloes having persistent corpus luteum. After an initial increase post-insemination, the plasma PGFM levels showed minor fluctuations. The assay was found to be sufficiently reliable and specific for estimation of PGFM levels in buffaloes. The standardization and validation of PGFM assay in buffalo opens the prospects of using PGFM levels as an indicator for reproductive health status monitoring in this species.

Animals↗
PubMed2003

Characterization and biological activity of gangliosides in buffalo milk.

Gangliosides (GS) were evaluated in Swiss cow's milk (SCM), Italian buffalo milk (IBM) and its serum, Pakistan buffalo colostrum (PBC), Pakistan buffalo mature milk (PBM), and Pakistan buffalo milk from rice-growing areas (PBR). Dairy GS were obtained from the Folch's upper (hydrophilic) and lower (lipophilic) extraction phases, respectively, and determined as lipid-bound sialic acid (LBSA) by colorimetry. Molar ratios of LBSA in the hydro- and lipophilic GS fractions were 52:48 to 79:21. Mature buffalo milk types had 40-100% more LBSA in the lipophilic GS fraction compared to SCM. Liquid PBC was higher in LBSA (24 nmol/g) compared to mature milk types (8-11 nmol/g). Thin-layer chromatography (TLC) and scanning densitometry showed distinct profiles of hydrophilic and lipophilic GS fractions. Lipophilic GS (but importantly not hydrophilic GS) from IBM and its serum decreased prostaglandin series 2 production by 75-80% in cultured human colonic epithelial cells exposed to tumor necrosis factor alpha (TNFalpha). Hydrophilic GD(3) and lipophilic GM(3) selectively bound rotavirus particles prepared from a rhesus strain and its mutant. A GS fraction in IBM showed a GM(1)-specific binding to cholera toxin subunit B (CTB). IBM serum (IBMS) was a rich source of LBSA (420 nmol/g proteins). In summary, improved methodology led to increased LBSA recovery and isolation of additional and bioactive milk GS. Human and Italian buffalo milk had similar CTB binding, and both had increased polysialo-GS compared to cows milk. The toxin binding properties of buffalo milk GS, and the anti-inflammatory activity of the lipophilized GS fraction could be important for developing innovative food applications, as well as the subject of future research.

Animals↗
PubMed1989

Characterization of buffalo-derived theilerial parasites with monoclonal antibodies and DNA probes.

The characteristics of intra-lymphocytic Theileria isolated from African buffalo and from cattle that were infected with buffalo-derived parasites were evaluated using anti-schizont monoclonal antibodies (mAbs) and DNA probes. Antigenic differences were revealed by the reactivities of 27 mAbs with the buffalo-derived parasites isolated from different animals. Antigenic diversity was also seen with Theileria-infected lymphoblastoid cell isolates taken from the lymph nodes and lambda gt11, showed specific hybridization to parasite DNA in Southern blots of restriction enzyme-digested, lymphoblastoid cells infected with buffalo-derived theilerial parasites. Genotypic differences between the buffalo-derived parasites were revealed by the restriction fragment length polymorphisms seen with hybridization of those probes to DNA from cloned and uncloned Theileria-infected cell lines. The evaluation of theilerial parasites derived from buffalo and from cattle which underwent typical T. p. lawrencei reactions, after being infected with buffalo-derived theilerial parasites, did not show any specific phenotypic or genotypic characteristics of these parasites that would distinguish them from T. p. parva and T. p. bovis parasites. The validity of these subspecies distinctions is discussed.

Animals↗
PubMed2002

Onset and sequence of RBA-band replication on the inactive X-chromosomes of cattle (Bos taurus L.), river buffalo (Bubalus bubalis L.) and goat (Capra hircus L.).

This study was undertaken to provide cytogenetic information about onset and sequence of RBA-band replication on the inactive X-chromosomes of cattle, river buffalo and goat. Blood cultures were synchronized overnight with thymidine after 48 hours of growth. The cell block was released with fresh medium and the cells allowed to grow in the presence of BrdU and H33258 for 1, 2, 4, 6, 8, 10, 12 and 14 hours, including 20 minutes colcemide. Results show that: (a) the onset of RBA-banding replication was 12 hours before mitosis in cattle and river buffalo, 14 hours in the goat; (b) the replication process was still 'on' in cattle and river buffalo one hour before mitosis, whereas it was 'off' in the goat; consequently the length of the G2 phase was less than one hour in cattle and river buffalo and one hour or slightly longer in the goat; (c) the first band undergoing replication was identified as band Xq31 in cattle, homologous to band Xq34-36 in river buffalo and Xq24 in the goat; (d) the second replicating band was the Xp22 in cattle, homologous to band Xq21 in river buffalo and Xq34 in the goat, respectively; (e) the sequence of RBA-band replication was quite similar between cattle and river buffalo, but reversed in the goat, due to the wide chromosomal rearrangements which differentiated the X-chromosome of Caprinae from that of Bovinae.

Animals↗