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H D Stone

Publications and source records attributed to H D Stone.

At least 37 records · Page 2Linked to original sources
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Response of congenitally immune chicks to viscerotropic velogenic Newcastle disease virus.

Separate groups of chicks of hen hyperimmune to viscerotropic velogenic Newcastle disease virus (VVNDV) were challenge-exposed to VVNDV by intraocular route at 1 day and 34 days old. Their response was evaluated by clinical symptoms, hemagglutination-inhibition (HI) titers, and virus isolations. Chicks exposed at 1 day old excreted VVNDV from the vent for up to 60 days; their active mean HI titers remained low (10-40); and deaths from VVNDV occurred early (5-16 days) and late (28-55 days). Chicks challenge-exposed at 34 days old excreted virus from the vent for 10 days; active HI titers developed quickly and remained high (891-1177); and deaths and signs of VVNDV occurred early (5-13 days).

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Simulation of maternal immunity by inoculation of immune yolk preparations into the yolk sac of 1-day-old chickens.

Yolk harvested from eggs laid by hens hyperimmunized with killed Newcastle disease virus (NDV) was inoculated into the yolk sac of 1-day-old specific-pathogen-free (SPF) chickens. Serum hemagglutination-inhibition antibody titers reached maximum levels 1 to 4 days after yolk inoculation and declined at a rate similar to that reported for naturally acquired maternal antibody. Expected levels of immune interference were observed when yolk-inoculated chickens were vaccinated with a conventional oil-emulsion NDV vaccine. These results show that yolk-sac inoculation with yolk antibody is a suitable approach for producing maternally immune chickens for laboratory studies.

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Evaluation of the efficacy of an oil-emulsion bacterin for protecting chickens against Salmonella enteritidis.

To assess the potential protective efficacy of a Salmonella enteritidis bacterin, an acetone-killed oil-emulsion vaccine was prepared from a phage type 13a S. enteritidis strain and administered subcutaneously to hens in two experiments. Hens were housed individually, and every other hen was vaccinated (at 23 weeks of age in one experiment and at 45 weeks in the other). A second (booster) bacterin injection was administered 6 weeks later in both experiments. Three weeks after the second vaccination, all hens were challenged with an oral dose of approximately 10(9) cells of a heterologous (phage type 14b) S. enteritidis strain. In both trials, S. enteritidis was isolated from fewer internal organs (spleens, ovaries, and oviducts) and pools of egg contents from vaccinated hens than from unvaccinated control hens. Vaccination did not, however, affect the percentage of hens that shed S. enteritidis in feces in either experiment.

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The preparation and efficacy of manually emulsified Newcastle disease oil-emulsion vaccines.

Experimental Newcastle disease oil-emulsion vaccines were prepared by manually shaking small quantities of antigen, surfactant, and mineral oil and compared with oil-emulsion vaccines made by the more conventional mechanical methods. Emulsion stability (prolonged emulsification of aqueous antigen) of the manually prepared vaccines was achieved by four different processes and tested for efficacy in broilers. White rock broilers were vaccinated at 3-5 weeks of age and bled at 1- to 2 week intervals thereafter for 8 weeks. Cumulative 8-week hemagglutination-inhibition (HI) mean titers (reciprocals) ranged from 15 to 250 for manually emulsified vaccines and from 18 to 240 for mechanically emulsified vaccines. Eight-week cumulative mean HI responses induced by manually emulsified vaccines were never significantly (P less than 0.05) lower than their specific mechanically emulsified comparison vaccines and were occasionally significantly higher. Highest HI titers were induced when hydrophile-lipophile balances of 7, 9, and 6 were used for the preparation of manually emulsified vaccines. In general, most of the vaccine emulsions were stable for more than 30 days. Vaccine efficacy, however, was not always diminished by poor emulsion stability. These results indicate that manual emulsification can be used for the production of oil-emulsion vaccines of high efficacy that can provide benefits over existing mechanical methods.

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Efficacy of experimental Newcastle disease water-in-oil oil-emulsion vaccines formulated from squalane and squalene.

Water-in-oil inactivated Newcastle disease oil-emulsion vaccines were formulated with the terpene oils squalane or squalene, or mixtures thereof, and injected into 4-week-old broilers. Vaccine efficacy based on hemagglutination-inhibition (HI) titers was comparable to that of control mineral oil vaccines. Tissue reaction to intramuscular injection of the terpene oil emulsion vaccines was greatly reduced 3 weeks post-vaccination compared with that of mineral oil-based vaccine. Viscosity of the terpene oil vaccines was satisfactory but increased three to four times that of mineral oil vaccine when the antigen phase volume increased from 5% to 20%.

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Immune response to oil-emulsion vaccines with single or mixed antigens of Newcastle disease, avian influenza, and infectious bronchitis.

Inactivated Newcastle disease virus (NDV), avian influenza virus (AIV), and infectious bronchitis virus (IBV) antigens were evaluated for immunological efficacy in monovalent and polyvalent vaccines. Vaccinated broilers were bled for hemagglutination-inhibition (HI) tests at 1- or 2-week intervals. Half of the chickens were challenged with the Largo isolate of velogenic viscerotropic (VV) NDV at 8 weeks post-vaccination, and the remainder were challenged with the Massachusetts 41 strain IBV at 9 weeks post-vaccination. Newcastle disease HI titers were reduced significantly (P less than 0.05) from those of monovalent control vaccine groups when IBV antigen was emulsified in mixtures with low (1-3x) concentrated NDV or NDV and AIV antigens. Avian influenza HI titers were significantly (P less than 0.05) lower than those of the control monovalent groups when highly concentrated NDV was part of the polyvalent vaccine. Infectious bronchitis HI titers were higher than those of control monovalent groups in 13 of 15 vaccine groups when IBV antigen was in polyvalent formulations. VV NDV challenge killed all non-NDV vaccinates and induced increased HI titers in NDV vaccinates but no morbidity or mortality. Sixty of 80 IBV vaccinates experienced a fourfold or greater HI titer increase following challenge. All non-IBV vaccinates seroconverted at 1 week post-challenge.

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Preparation of inactivated oil-emulsion vaccines with avian viral or Mycoplasma antigens.

The influence of the composition of water-in-oil emulsions on their physical characteristics was determined by preparing experimental emulsions with various water-to-oil ratios and various emulsifiers. Emulsions containing Tween 80 in the aqueous phase and Arlacel A or Arlacel 80 in the oil phase were lower in viscosity than emulsions containing only an oil-phase emulsifier. Viscosity decreased as the concentration of oil increased. Oil-emulsion vaccines prepared with aqueous- and oil-phase emulsifiers had low viscosity, were stable for more than 12 weeks at 37 C, and induced a marked primary antibody response in chickens.

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Efficacy of oil-emulsion vaccines prepared with pigeon paramyxovirus-1, Ulster, and La Sota Newcastle disease viruses.

Three strains of avian paramyxovirus-1 virus (PMV-1) were used to prepare four experimental monovalent oil-emulsion vaccines. A pigeon PMV-1 isolate (PPMV-1) and the Newcastle disease virus strains La Sota and Ulster were used to prepare four pools of beta-propiolactone-inactivated allantoic fluid for the vaccines. Groups of susceptible white rock chickens and racing homing pigeons were vaccinated subcutaneously with one of the vaccines, and their serologic responses were determined using the hemagglutination-inhibition (HI) test at frequent intervals up to 9 weeks postvaccination. Pigeons were challenged after 10 weeks with a virulent PPMV-1 isolate given intravenously, observed for signs of disease for 5 weeks, and then tested for secondary serologic HI responses. The HI responses were measured using the three strains of virus as HI test antigens. The titers were generally greater when the hemagglutination antigen used in the test was homologous with the antigen used to prepare the vaccine. All vaccines protected pigeons against morbidity and death but not against infection with the challenge virus. The shedding of PPMV-1 challenge virus from PPMV-1 vaccinates was greatly reduced 6 days after challenge.

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Efficacy of avian influenza oil-emulsion vaccines in chickens of various ages.

An experimental avian influenza (AI) oil-emulsion vaccine was formulated with 1 part inactivated A/turkey/Wisconsin/68 (H5N9) AI virus emulsified in 4 parts oil. Broilers were vaccinated subcutaneously (SC) either at 1 or 3 days old or at 4 or 5 wks old. Commercial white leghorn (WL) layers were vaccinated SC at 12 and 20 wks old or at only 20 wks old. Maximum geometric mean hemagglutination-inhibition titers postvaccination (PV) were 1:86-1:320 for broilers, 1:597 for twice-vaccinated layers, and 1:422 for once-vaccinated layers. Ninety to 100% of vaccinated broilers were protected against death and morbidity when challenged with highly pathogenic A/chicken/Penn/83 (H5N2) AI virus 4 weeks PV, and all were protected when challenged 8 wks PV. All controls and most vaccinates were infected by challenge virus, and 90-100% of controls died or exhibited clinical signs. Vaccinated commercial pullets were protected against morbidity, death, and egg-production decline at either peak of lay (25 wks old) or at 55 wks old. All unvaccinated controls became morbid or died, and egg production ceased 72 hours after challenge. The 0.5-ml vaccine dose was determined to contain 251 and 528 mean protective doses (PD50S) in 4-wk-old and 1-year-old SPF WL chickens, respectively, challenged 4 wks PV.

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Optimization of hydrophile-lipophile balance for improved efficacy of Newcastle disease and avian influenza oil-emulsion vaccines.

Preparations of inactivated Newcastle disease (ND) and avian influenza (AI) oil-emulsion vaccines with surfactant hydrophile-lipophile-balance (HLB) values between 4.3 and 9.5 were evaluated for their efficacy in broiler-type white rock chickens. Chickens were vaccinated at 3-4 weeks of age and bled at 2-week intervals over 8 weeks. Post-vaccinal hemagglutination-inhibition (HI) geometric mean titers (reciprocals) ranged from 197 to 485 for ND vaccines and from 184 to 1040 for AI vaccines. Based on the HI response, an HLB value of 7.0 induced the greatest stimulation of antibody titers. Ten percent surfactant in the oil phase of the vaccines induced maximum titers at this HLB. The oil:aqueous ratios of the vaccines did not greatly influence the overall serologic response when the vaccines had an HLB of 7.0. These results indicate that manipulating surfactant HLB values of OE vaccine may maximize the HI response in broilers.

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Effect of thimerosal concentration on the efficacy of inactivated Newcastle disease oil-emulsion vaccines.

Different quantities of the preservative thimerosal in inactivated Newcastle disease oil-emulsion vaccines were tested to determine the influence on the hemagglutination-inhibition (HI) response of broilers. The effect of thimerosal was measured in vaccines that had been stored for 1, 21, and 52 weeks; HI serology was conducted at 2, 4, and 6 weeks after vaccination. Mean HI titers 4 weeks after vaccination decreased at a significant rate (P less than or equal to 0.001) with increasing concentrations of thimerosal. HI titers 4 weeks after vaccination with 1-week-old vaccine were significantly (P less than or equal to 0.05) higher than those after vaccination with 52-week-old vaccine at all thimerosal concentrations tested. Titers were also significantly higher (P less than or equal to 0.05) after vaccination with 1-week-old vaccine than after vaccination with 21-week-old vaccine at all thimerosal concentrations below about 8.25 mg/ml of antigen. Thimerosal at the levels recommended in commercial vaccines does not significantly decrease vaccine efficacy.

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Determination of hemagglutination activity recovered from oil-emulsion Newcastle disease vaccines as a prediction of efficacy.

Hemagglutination (HA) activity was recovered from the aqueous phases of commercially and experimentally prepared oil-emulsion (OE) Newcastle disease virus (NDV) vaccines using two methods: aqueous partition and freeze-thaw. Quantitation of the HA activity retrieved by the aqueous partition technique was directly related to the degree of protection conferred to chickens against a strain of velogenic viscerotropic (VV) NDV in nine of the ten vaccines analyzed. One of the ten vaccines yielded high levels of retrieved HA activity but induced low hemagglutination-inhibition (HI) antibody levels and low levels of protection. Therefore, the retrieval and quantitation of HA activity from OE NDV vaccines using the partition technique provides a useful adjunct to vaccine testing methods that doesn't require vaccination and challenge trials. The freeze-thaw method did not yield measurable HA titers for all vaccines with high efficacy, so its use should be restricted to only those vaccines for which it has been demonstrated to be suitable.

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Influence of formulation on the efficacy of experimental oil-emulsion Newcastle disease vaccines.

Twenty-one experimental oil-emulsion vaccines with different emulsifier contents, aqueous-to-oil ratios, and antigen concentrations were compared by immunization of 4-week-old chickens. Vaccines that contained oil-phase (Arlacel 80) and aqueous-phase (Tween 80) emulsifiers induced 2-to-4-fold higher hemagglutination-inhibition titers than vaccines with only the oil-phase emulsifier. The emulsion vaccines containing both emulsifiers were also more stable at 37 C and less viscous than those containing only the oil-phase emulsifier. Vaccines that had different aqueous-to-oil ratios and contained different quantities of allantoic-fluid antigen (1.2% to 50% of the vaccine volume) induced similar protection against challenge, but hemagglutination-inhibition titers were proportional to the amount of antigen added. Vaccines that had different aqueous-to-oil ratios but contained equal amounts of antigen induced similar hemagglutination-inhibition titers and similar protection against challenge.

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Comparison of three experimental inactivated oil-emulsion Newcastle disease vaccines.

White Leghorn pullets vaccinated previously with live Newcastle disease virus (NDV) were revaccinated at point-of-lay with 3 experimental oil-emulsion (OE) Newcastle disease vaccines to determine whether immunogenicity of OE vaccines is influenced by emulsion composition differences. The vaccines used were formulated differently but contained equal amounts of inactivated NDV in stable, low-viscosity, water-in-oil emulsions. Serological responses to the vaccines differed (P less than 0.01), but 55 of 56 OE vaccinates remained clinically normal following challenge exposure with viscerotropic velogenic NDV 28 or 44 weeks after revaccination; 4 of 21 nonrevaccinated controls and all of 20 nonvaccinated challenge controls were clinically affected or died.

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Evaluation of the efficacy of oil-emulsion bacterins for reducing fecal shedding of Salmonella enteritidis by laying hens.

Two replicate experiments were conducted to test the efficacy of two different Salmonella enteritidis oil-emulsion bacterins (an experimentally prepared acetone-killed vaccine and a commercially available vaccine) for protecting laying hens against intestinal colonization following oral exposure to S. enteritidis. Each vaccine was administered twice (4 weeks apart), and all hens were challenged with 10(8) cells of a nalidixic-acid-resistant S. enteritidis strain 2 weeks after the second vaccination. Fecal samples from vaccinated and unvaccinated control hens were cultured at three weekly intervals post-challenge to determine the incidence of intestinal colonization and the numbers of S. enteritidis shed into the environment. Both vaccines significantly reduced the incidence of intestinal colonization (P < 0.05) and the mean number of S. enteritidis cells shed in the feces (P < 0.01) at 1 week post-challenge. However, the degree of protection afforded by vaccination was only partial, as more than half of the vaccinated hens still shed substantial numbers of S. enteritidis. If used in conjunction with other flock sanitation and infection-monitoring strategies, vaccination with bacterins could potentially reduce the overall level of environmental contamination and thereby also reduce the horizontal transmission of S. enteritidis within and between laying flocks.

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