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Biomedical subjects

B M Bindon

Publications and source records attributed to B M Bindon.

At least 19 recordsLinked to original sources

Effects of various adjuvants on efficacy of a vaccine against Streptococcus bovis and Lactobacillus spp in cattle.

OBJECTIVE: To determine efficacy of vaccines incorporating QuilA, alum, dextran combined with mineral oil, or Freund adjuvant for immunization of feedlot cattle against Streptococcus bovis and Lactobacillus spp. ANIMALS: 24 steers housed under feedlot conditions. PROCEDURE: Steers were randomly assigned to 4 experimental groups and a control group. Animals in experimental groups were inoculated on days 0 and 26 with vaccines containing Freund adjuvant (FCA), QuilA, dextran combined with mineral oil (Dex), or alum as adjuvant. Serum anti-S bovis and anti-Lactobacillus IgG concentrations were measured, along with fecal pH, ruminal fluid pH, and number of S bovis and Lactobacillus spp in ruminal fluid. RESULTS: Throughout the study, serum anti-S bovis and anti-Lactobacillus IgG concentrations for animals in the Dex, QuilA, and alum groups were similar to or significantly higher than concentrations for animals in the FCA group. Serum anti-S bovis and anti-Lactobacillus IgG concentrations were significantly increased on days 26 through 75 in all 4 experimental groups, and there was a linear relationship between anti-S bovis and anti-Lactobacillus IgG concentrations. For animals in the QuilA and Dex groups, mean pH of feces throughout the period of experiment were significantly higher and numbers of S bovis and Lactobacillus spp in ruminal fluid on day 47 were significantly lower than values for control cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that immunization of feedlot steers against S bovis and Lactobacillus spp with vaccines incorporating Freund adjuvant, QuilA, dextran, or alum as an adjuvant effectively induced high, long-lasting serum anti-S bovis and anti-Lactobacillus IgG concentrations. Of the adjuvants tested, dextran may be the most effective.

Adjuvants, Immunologic↗

Synchronization of estrus and fertility in zebu beef heifers treated with three estrus synchronization protocols.

The effects on estrus and fertility of 3 estrus synchronization protocols were studied in Brahman beef heifers. In Treatment 1 (PGF protocol; n=234), heifers received 7.5 mg, i.m. prostianol on Day 0 and were inseminated after observed estrus until Day 5. Treatment 2 (10-d NOR protocol; n = 220) consisted of norgestomet (NOR; 3 mg, s.c. implant and 3 mg, i.m.) and estradiol valerate (5 mg, i.m.) treatment on Day -10, NOR implant removal and 400 IU, i.m. PMSG on Day 0, and AI after observed estrus through to Day 5. Treatment 3 (14-d NOR+PGF protocol; n = 168) constituted a NOR implant (3 mg, sc) on Day -14, NOR implant removal on Day 0, PGF on Day 16, and AI after observed estrus through to Day 21. All heifers were examined for return to estrus at the next cycle and inseminated after observed estrus. The heifers were then exposed to bulls for at least 21 d. During the period of estrus observation (5 d) after treatment, those heifers treated with the PGF protocol had a lower (P<0.01) rate of estrual response (58%) than heifers treated with the 10-d NOR (87%) or 14-d NOR+PGF (88%) protocol. Heifers treated with the 10-d NOR protocol displayed estrus earlier and had a closer synchrony of estrus than heifers treated with either the PGF or the 14-d NOR+PGF protocol. Heifers treated with the 14-d NOR+PGF protocol had higher (P<0.05) conception and calving rates (51 and 46%) to AI at the induced estrus than heifers treated with the PGF (45 and 27%) or the 10-d NOR (38 and 33%) protocol. Calving rate to 2 rounds of AI was greater (P<0.05) for heifers treated with the 14-d NOR-PGF (50%) protocol than heifers treated with the 10-d NOR (38%) but not the PGF (43%) protocol. Breeding season calving rates were similar among the 3 protocols. The results show that the 14-d NOR+PGF estrus synchronization protocol induced a high incidence of estrus with comparatively high fertility in Brahman heifers.

Animal Husbandry↗

Increased ovulation rate in Merino ewes immunized against small synthetic peptide fragments of the inhibin alpha subunit.

Four experiments were carried out in Merino ewes during a period of 4 years to determine the long-term effects of immunization against different synthetic peptides mimicking the amine terminal of the alpha subunit of porcine inhibin. Peptides were conjugated to human serum albumin and 100-200 micrograms emulsified in Freund's complete adjuvant for the primary immunization. Usually two booster injections were given at monthly intervals with 50-100 micrograms conjugated peptide using either incomplete Freund's adjuvant or Montanide:Marcol. In some experiments a further immunization was carried in the next year. Blood samples were taken 10 days after each immunization, during the luteal phase, for estimation of gonadotrophin concentrations and determination of inhibin antibody titres. One day after blood sampling cloprostenol was used to induce luteolysis and laparoscopy was performed in the subsequent oestrous cycle. Immunization of ewes with synthetic peptides 1-32, 1-26, 7-26 and 8-30 resulted in large increases in the ovulation rate (OR). An approximately two-fold increase in OR was observed following the first booster immunization with these peptides and a three- to five-fold increase after the second booster immunization. Immunization with these large peptides resulted in a sustained increase in OR for a period of at least 1 year after the second booster immunization. Of the shorter peptides, peptides 10-26 and 13-26 gave a reasonable ovulatory response, although it was more difficult to obtain a response with peptides 1-16, 8-22, 13-25, 8-19 and 10-19; peptides 7-13 and 1-6 gave no response (but were examined for one breeding season only). The smaller peptides led to lower inhibin antibody titres that were not necessarily associated with increased follicle-stimulating hormone (FSH) or OR. More intensive blood sampling in one experiment showed that following primary immunization against peptide 1-32 there was a transient increase in plasma FSH, which did not lead to an increased OR. Moreover, a prolonged period of raised FSH after the first booster was significantly correlated with increased OR. In these animals antibody titres were only slightly increased after primary immunization, but after the first booster immunization higher titres were observed that were significantly correlated with trough FSH values and the subsequent OR. These results are interpreted as showing that (1) to obtain an increase in OR peptides 1-32, 1-26 and 7-26 are suitable as immunogens; (2) smaller peptides are less reliable, often require multiple injections, and the response may be delayed; and (3) an extended period of raised plasma FSH is needed to give a large ovulatory response.

Animals↗

Immunological manipulation of ovulation rate for twinning in cattle.

Unlike in sheep, in which immunization against androstenedione causes mild and reasonably controlled increased ovulation rate, in similar studies cattle showed highly variable responses ranging from increased ovulation rate and fertility through to anovulation/anoestrous or superovulation. As a consequence, interest in manipulation of ovulation rate through this approach has declined and is now focused on immunological manipulation of endogenous inhibin following successful studies in sheep. Studies have concentrated on developing a prototype inhibin-based vaccine to be used for twinning in the Australian beef industry. The prototype vaccine (with recombinant ovine inhibin-alpha.3 fusion protein and Montanide:Marcol adjuvant) has proved to be very potent and control of the degree of ovarian stimulation has not been possible. The proportion of cattle with increased ovulation rate after inhibin immunization is affected by timing of booster vaccination within the ovarian cycle, time after vaccination, vaccine formulation and possibly genotype. Physiological studies show that cattle responding to the inhibin vaccine have increased plasma inhibin binding of native bovine inhibin, high plasma FSH concentrations, greater numbers of large (> or = 8 mm) follicles and fewer small (< 5 mm) follicles during the preovulatory wave of follicular development compared with control or non-responding animals. Significant correlations among the response parameters (i.e. inhibin binding, plasma FSH concentrations, number of large follicles and ovulation rate) have been demonstrated. The results indicate that greater understanding of the various processes of folliculogenesis will be necessary to achieve a controlled increase in ovulation rate in cattle.

Animals↗

Fertility of superovulated ewes after intrauterine or oviducal insemination with low numbers of fresh or frozen-thawed spermatozoa.

Two experiments investigated the effects of number of spermatozoa inseminated and time and site of insemination of fresh and frozen-thawed semen on fertility in Merino ewes treated with intravaginal progestagen pessaries and a combination of pregnant mare serum gonadotrophin and follicle stimulating hormone. In Experiment 1, ewes were inseminated in the uterus or oviducts with totals of 10(4), 10(5), 10(6) or 10(7) fresh spermatozoa 44 or 68 h after pessary removal. Ova recovered 48 h later were classified as fertilized if they had cleaved. Proportion of fertile ewes (ewes with fertilized ova per ewes inseminated) and fertilization rate of ova (ova fertilized per ova recovered) were higher for inseminations 44 h (63% and 45%) than 68 h (38% and 19%) after pessary removal (P < 0.001). More ewes were fertile after oviducal (61%) than after intrauterine insemination (39%; P < 0.05), and with high (10(6) and 10(7)) than with low (10(4) and 10(5)) doses of spermatozoa for intrauterine (54% v. 24%; P < 0.05) but not for oviducal inseminations (63% v. 59%). Fertilization of ova was better after oviducal than after intrauterine inseminations (44% v. 22%; P < 0.001), and with high than with low doses of spermatozoa (45% v. 21%; P < 0.001). In Experiment 2, ewes were inseminated with 10(5), 5 x 10(5) or 10(6) total fresh or frozen-thawed spermatozoa in the uterus or oviducts, 44 h after pessary removal.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Active immunization of Merino ewe lambs with recombinant bovine alpha inhibin advances puberty and increases ovulation rate.

Ewe lambs (n = 24-25) were immunized at 3, 7 and 15 weeks of age with recombinant bovine alpha-inhibin (rec inhibin) or with bovine monoclonal antibody purified inhibin (bMPI) obtained by immunochromatography from bovine follicular fluid or with adjuvant alone (control). Antibodies in the plasma of the lambs immunized with the inhibin preparations bound to iodinated 31 kDa bovine inhibin. Binding was minimal after the primary immunization, increased after each booster immunization and remained elevated until at least 45 weeks of age (29% for rec inhibin and 11% for bMPI). Of the group treated with rec inhibin, 10 ovulated as lambs (control 0/22; bMPI 3/21) and onset of overt oestrous activity (as hoggets) was advanced (P < 0.05) by 17 days in lambs immunized with rec inhibin. As hoggets, the ovulation rate was greater (P < 0.01) in the rec inhibin immunized lambs (4.41 +/- 0.67) than in the control animals (1.27 +/- 0.15) but not in the bMPI-treated lambs (1.40 +/- 0.16). After a further immunization at 17 months of age, however, ovulation rate increased (P < 0.01) in the bMPI-immunized group (3.40 +/- 0.47) but was unchanged in the lambs immunized with rec inhibin (2.80 +/- 0.52) and those in the control group (1.15 +/- 0.08). There were no effects of immunization on plasma concentrations of either follicle stimulating hormone (FSH) or luteinizing hormone (LH). The initial smaller response seen with the bMPI may be due to either the presence of compounds other than inhibin in such preparations or the small absolute amount of inhibin injected.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Immunization of merino ewes with a synthetic inhibin peptide or with preparations obtained from bovine and porcine follicular fluids by immunoaffinity chromatography result in different effects on ovulation rate and on plasma gonadotrophin concentrations.

Ewes were immunized with either a synthetic peptide (peptide 1-32) that has an amino acid sequence identity with the first 32 amino acids at the amino terminal of the alpha-subunit of porcine inhibin, or with bovine or porcine monoclonal antibody purified inhibin (bMPI and pMPI respectively), obtained by immunochromatography from follicular fluids. The peptide 1-32 was conjugated to albumin before use. Peptide 1-32 and bMPI increased ovulation rate and number of follicles (greater than or equal to 5 mm diameter). Although bMPI increased plasma FSH concentration the peptide did not. pMPI had no effect on ovarian activity but markedly elevated both plasma FSH and LH concentrations. The plasma LH concentration was lowered in ewes immunized with peptide 1-32. It appears, therefore, that ovulation rate can be increased following increased plasma FSH concentrations at luteolysis or in the absence of such an increase. Conversely, greatly increased plasma gonadotrophin concentrations at luteolysis (pMPI) were not followed by an increase in ovulation rate. Antibodies in the plasma of ewes immunized with peptide 1-32 and bMPI bound to iodinated synthetic human inhibin alpha-chain 6-30 peptide. The results suggest that ovulation rate is at least partly determined by intraovarian factors.

Animals↗

Ovarian follicular dynamics in lines of sheep (Finn, Merinos) selected on ovulation rate.

Ewes from selected lines of sheep from each of two breeds, Finns (high ovulation rate, low ovulation rate and control lines with respective ovulation rates of 5.4, 2.7 and 3.3) and Merinos (T Merinos selected for increased ovulation rate and control Merinos with respective ovulation rates of 1.9 and 1.2) were used to examine how selection to alter ovulation rate had altered follicle development. Ovarian antral follicles were counted, measured, classified as nonatretic or atretic (more than five pyknotic bodies). The growth of ovulatory follicles in vivo, followed by repeated follicle ink marking, also was compared in the three lines of Finns. Regardless of breed, ewes selected for high ovulation rate had a similar number of antral follicles and a similar extent of atresia compared with their controls. Alterations induced by selection were located in the last stages of folliculogenesis. T Merinos exhibited a lower proportion of atretic follicles among follicles greater than 3 mm and a larger diameter of the largest healthy follicle when preovulatory follicles were excluded. High-line Finn ewes recruited more follicles, which produced smaller preovulatory follicles, each containing a smaller number of granulosa cells compared with either the low- or control-line ewes. Hence, physiological selection for high ovulation rate raised it by different methods in Merino than in Finn ewes.

Animals↗

Increase in ovulation rate in Merino ewes after active immunization with inhibin preparations obtained by immunoaffinity chromatography.

Ewes were immunized with four inhibin preparations of increasing purity obtained from bovine follicular fluid by affinity chromatography. The two purest immunogens, which had 69 and 174 ng inhibin micrograms-1 protein (expressed in terms of 32 kDa ovine inhibin), increased ovulation rate by 240 and 320% respectively following two injections, and gave a transient increase in plasma FSH concentration at the same time. Plasma antibodies recognizing native 125I-31 kDa bovine inhibin appeared at the same time, and, together with the increased ovulation rate, persisted after the injections ceased. Ovulation rate and antibody titres were raised by the less pure immunogens, but to a smaller extent, and the increases did not persist. It was concluded that the increased ovulation rate was due to induced inhibin antibodies, acting at least in part through the consequent rise in plasma FSH concentration.

Analysis of Variance↗

Induction of ovulation in chronically hypophysectomized Booroola ewes.

Booroola Merino ewes, with (F+; N = 17) and without (++; N = 13) a copy of the fecundity gene were hypophysectomized and 6 weeks later were given an i.m. injection of PMSG (high, medium or low dose) followed by hCG. The induced ovulation rates were observed laparoscopically. Ovulation rates were significantly higher (P less than 0.01) in Booroola F+ ewes than in ++ ewes (8.00 +/- 1.66 s.e.m. vs 3.62 +/- 1.10 respectively). This suggests that the high fecundity of the Booroola ewe may be due primarily to ovarian rather than pituitary factors.

Animals↗

Characterization of LH and testosterone responses to intramuscular injection of GnRH in tropical postpubertal bulls.

Five Zebu x British crossbred bulls 17 months of age and of uniform liveweight (320+/-3 kg) were used to study testosterone responses to single intramuscular doses of exogenous gonadotropin-releasing hormone (GnRH). The eight dose levels used were 0, 31.25, 62.5, 125, 250, 500, 1000, and 2000 ng GnRH/kg live weight. Plasma samples for hormone responses were collected at 30-minute intervals from zero to three hours and at one-hour intervals from three to seven hours postinjection. Luteinizing hormone (LH) and testosterone responses were measured as peak heights or as areas under response curves. Increasing the dosage of GnRH increased the time to reach the peak LH response, the height and duration of the response, and the area under the response curve. The maximum LH peak height was reached by the 1 mug/kg dose. In contrast to LH, testosterone responses reached the same peak heights (two hours postinjection of GnRH) for all doses of GnRH. The only effect of increased dosage was to increase the duration of response. Testosterone responses showed repeatable differences (P<0.01) between animals, but LH responses did not. It was demonstrated that the testosterone status of bulls can be accurately assessed by simply measuring testosterone in a single plasma sample collected two to three hours after the intramuscular injection of 100 mug or more (dose unimportant) of GnRH per bull.

Journal Article↗

Reproductive characteristics of lambs actively immunized early in life with inhibin-enriched preparations from follicular fluid of cows.

Active immunization of Merino and crossbred ewe lambs early in life with a partly purified inhibin derived from bovine follicular fluid (bFF) advanced their puberty. This occurred whether the lambs were immunized from 3 weeks of age or from 9 weeks of age or just 3 times between 3 and 9 weeks. However, the effect was more obvious with multiple injections starting at 3 weeks of age. The ovulation rate of the immunized lambs was significantly higher than that of the control ewes. When lambs were subjected to multiple immunizations the increased ovulation rate persisted in Merinos for at least 1 year after immunization ceased. The lambs injected 3 times only (at 3, 6 and 9 weeks of age) did not show any increase in ovulation rate. Plasma concentrations of FSH and LH measured in blood samples taken 1 week after immunization were not significantly different between the treatment groups. Daily sperm output in the urine and testis diameter were measured in ram lambs from the same breeds and flocks as the ewe lambs. Immunization with the inhibin preparation from 3 weeks of age resulted in a significant increase in daily sperm output (4255 +/- 701 vs 2344 +/- 344 x 10(6), P less than 0.01) and testis diameter (5.21 +/- 0.3 vs 4.33 +/- 0.2 cm, P less than 0.05) in Merino rams examined in the non-breeding season when the rams were aged 23 months. Although the same trend was seen in the following year the differences were not significant. Immunization from 9 weeks of age had no effect. A similar increase in daily sperm output was seen (at 20 and 25 months of age) in crossbred rams immunized from 3 weeks of age but the difference was not significant. Plasma concentrations of FSH (but not LH) in samples from rams immunized from 3 weeks of age were significantly higher than those of control rams at 7 and 60 weeks of age in Merino rams and at 23 and 31 weeks of age in crossbred rams. Plasma FSH concentrations at 21 and 26 weeks of age in the Merino rams immunized from 3 weeks of age were higher than those of control lambs, and these increases preceded a significant (P less than 0.05) increase in testicular diameter at 30 weeks of age (4.99 +/- 0.2 vs 4.37 +/- 0.2 cm). These results suggest that active immunization early in life with an inhibin-enriched fraction from bFF advances puberty in ewe lambs.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

The half-life of follicle-stimulating hormone in ovary-intact and ovariectomized booroola and control merino ewes.

To determine whether the high ovulation rate of the Booroola Merino ewe could be explained by FSH metabolism we have tested the proposition that FSH may have a longer half-life in the plasma of Booroola Merino ewes than in control ewes. The half-life of plasma FSH was determined by removal of the pituitary gland, to abolish FSH secretion into the peripheral circulation, and monitoring by repeated blood sampling the subsequent decline in plasma FSH concentrations. The half-life of FSH was similar in Booroola (103 +/- 14 (s.e.m.) min, N = 8) and control (116 +/- 8 min, N = 9) ewes. However, when ewes that had been ovariectomized at least 6 months earlier were hypophysectomized, the half-life of FSH was increased from 110 + 8 min in ovary-intact ewes (N = 11) to 1101 +/- 49 min (N = 6) (P less than 0.001) with no difference between the two Merino strains. We conclude that changes in the circulating half-life of FSH do not account for the high fecundity of the Booroola but that ovariectomy can alter the half-life of FSH secreted by the pituitary gland.

Animals↗

Increase in ovulation rate after immunization of Merino ewes with a fraction of bovine follicular fluid containing inhibin activity.

Affinity chromatography of bovine follicular fluid using Matrix gel Red A resulted in a 20-fold increase in inhibin-like specific activity assessed in a mouse ovulation inhibition test system. When this material was emulsified with Freund's adjuvant and injected into adult Merino ewes their mean ovulation rate was increased from 1.2 to 2.3 (P less than 0.01). Follicles of diameter greater than or equal to 3.5 mm and 2-3.4 mm were also increased (4.33 vs 2.25 and 5.39 vs 2.44 per ewe respectively). The ovulation rate response was variable and transient. Length of oestrous cycles, number of granulosa cells per follicle and seasonal oestrous patterns were not affected. Plasma from the immunized ewes contained antibodies to the immunogen and reversed the ovulation-inhibiting effects of bovine follicular fluid in mice.

Animals↗

Suppression of ovine plasma FSH by bovine follicular fluid: neutralization by plasma from ewes immunized against an inhibin-enriched preparation from bovine follicular fluid.

Adult Merino ewes were immunized against an inhibin-enriched preparation (bFFI) obtained by affinity chromatography of bovine follicular fluid (bFF). Plasma was obtained in early luteal phase from these ewes and from control ewes immunized against bovine serum albumin. Ten months after ovariectomy the plasma concentration of FSH, but not LH, in control ewes was decreased by four s.c. injections of 8 ml bFFI (17,500 units inhibin/injection). There was no decrease in plasma concentrations of FSH or LH in immunized ewes with the same dose of bFFI. In a second study with long-term ovariectomized ewes, four injections of 20 ml plasma from the immunized ewes significantly reduced the decrease in FSH concentration caused by four injections of steroid-free bFF (2500 units inhibin/injection) in comparison with similar ewes injected with plasma from control ewes. These results show that the plasma of ewes immunized against bFFI contains substances, presumably antibodies, which neutralize the FSH-suppressive effects of bFF and bFFI in ovariectomized ewes.

Animals↗