Mycoplasma Gallisepticum Vaccine Research Articles

A Comparative Study of Live Attenuated F Strain–Derived Mycoplasma gallisepticum Vaccines

A Comparative Study of Live Attenuated F Strain–Derived Mycoplasma gallisepticum Vaccines

Evaluation of Mycoplasma gallisepticum K-Strain as a Live Vaccine in Chickens

We evaluated the pathogenicity of three live Mycoplasma gallisepticum (MG) vaccine candidates by infection via aerosol of 3-wk-old chickens with log phase broth cultures (trial 1). Two of the candidates (K3020 and K4649A) colonized only 10% and 20% of the chickens, respectively, unlike K2101 (K-strain), which was reisolated from all of the vaccinated chickens tested. K-strain inoculation did not result in significant air sac or tracheal lesions in chickens at 10 and 39 days postinfection (P < or = 0.05). The efficacy of K-strain as a live vaccine was evaluated in trial 2, by challenge of vaccinated chickens with virulent R-strain via aerosol at 6 wk postvaccination. K-strain vaccination resulted in significant protection from air sac and tracheal lesions (P < or = 0.05). The K-strain was further investigated to evaluate transmissibility (trial 3), colonization and persistence of infection following aerosol administration (trial 4), genetic and phenotypic stability following back passage through chickens (trial 5), and vertical transmission (trial 6). The K-strain had a low rate of horizontal transmission; it remained primarily in the respiratory system of inoculated birds and persisted in the upper respiratory tract for the duration of the trial 4 (5 mo). There was no increase in virulence of K-strain when it was back passaged five times through chickens, and no vertical transmission of K-strain was detected. K-strain showed great potential as a safe and effective live MG vaccine.

Serologic Response of Roosters to Gradient Dosage Levels of a Commercially Available Live F Strain-Derived Mycoplasma gallisepticum Vaccine Over Time

Spray application is a commonly used, time- and labor-efficient means to deliver live Mycoplasma gallisepticum (MG) vaccine to laying hens in commercial production facilities. The dosage of vaccine received by spray-vaccinated birds can vary due to variation in the spray plume and the vaccine suspension droplet trajectory. In this study, a total of 48 Hy-Line W-36 males were placed individually in isolation units following eye-drop application of gradient levels (1 x, 10(-1) x, 10(-2) x, 10(-3) x, 10(-4) x, 10(-5) x, 10(-6) x, and unvaccinated control) of the MG vaccine. The determined titer associated with a 1 x dose was 2 x 10(6) colony-forming units/dose. Serologic response was assessed weekly following vaccination via serum plate agglutination (SPA) for weeks one through seven postvaccination (p.v.). In addition, immunologic response was assessed at 5, 6, and 7 wk p.v. via MG enzyme-linked immunosorbent assay (ELISA). As indicated by SPA analyses, a 1 x dose of vaccine resulted in 100% seroconversion, and dose levels of 10(-1) x and 10(-2) x resulted in 75% and 37.5% seroconversion, respectively, at 6 wk p.v. The MG ELISA results at 6 wk p.v. demonstrated immunologic responses in 100%, 57.1%, and 28.6% of the 1 x, 10(-1) x, and 10(-2) x dosed birds, respectively. The lower dosage levels of 10(-3) x, 10(-4) x, 10(-5) x, and 10(-6) x did not elicit a response from any bird at 6 wk p.v. Utilizing the SPA data, a logistic regression model was used to determine the relationship between dosage level and seroconversion rate (R2 = 0.999 with a standard error of prediction of 1.6%). The model predicted a required effective dosage of 0.26 x for 90% seroconversion at 6 wk p.v. under test conditions.

Serologic Response of Roosters to Gradient Dosage Levels of a Commercially Available Live F Strain-Derived Mycoplasma gallisepticum Vaccine Over Time

Serologic Response of Roosters to Gradient Dosage Levels of a Commercially Available Live F Strain-Derived Mycoplasma gallisepticum Vaccine Over Time

Development and immunogenicity of recombinant GapA+Mycoplasma gallisepticum vaccine strain ts-11 expressing infectious bronchitis virus-S1 glycoprotein and chicken interleukin-6

Mycoplasma gallisepticum (MG) is a major pathogen of poultry that causes chronic respiratory disease in chickens and infectious sinusitis in turkeys. A live attenuated vaccine, ts-11, has been used for the control of MG in several countries. The efficacy of this vaccine is highly dose dependent and the flock antibody response is weak. To improve the functionality of the vaccine and investigate its potential as a delivery vector for foreign antigens and immunomodulatory proteins, we developed a derivative of ts-11 expressing infectious bronchitis virus-S1 glycoprotein (IBV-S1) and releasing chicken interleukin-6 into the extracellular milieu (MG ts-11 C3 (+CS)) using a transposon-based delivery vector. Following administration of MG ts-11 C3 (+CS) to chickens by eye-drop, an antibody response to MG and IBV-S1, as determined by the rapid serum agglutination test (RSA) and Western blotting, respectively, could be detected. Birds inoculated with the recombinant vaccine had significantly enhanced weight gain and were partially protected against damage by pathogenic IBV. These results indicate that the ChIL-6 released by MG ts-11 C3 (+CS) may have had a non-specific effect on growth rate. They also suggest that ts-11 is a promising vaccine vector, capable of delivering heterologous protective antigens, and may also provide non-specific benefits when engineered to express immunomodulatory proteins. With some improvements in the expression system, it could be used to induce a targeted immune response against specific mucosal pathogens, and co-expression of several antigens would allow development of a novel multivalent vaccine.

Eye Surface Area and Dosage Rates for Spray Vaccination

Spray application of Mycoplasma gallisepticum (MG) vaccines is a labor- and time-saving means of mass vaccination of layer chickens. Recent assessment of spray characteristics of nozzles commonly used to apply MG vaccine in layer chicken operations has shown that the amount of respirable droplets (< 5 microm) is negligible. Topical application of vaccine onto the eye surface has been suggested as a route of vaccination, but no estimates of vaccine load delivered via spray application were found in the literature. Estimates of eye surface area were developed using digital imaging; 24 layer pullets were used for analysis, and the mean eye surface area, corrected for corneal curvature, was found to be 0.609 cm2. This surface area was then used to estimate vaccine load for commercially available live MG vaccine sprayed through popular nozzles. Less than 3000 colony-forming units can be expected for direct deposition onto the surface of an eye.

A Chronicle of Serologic Response in Commercial Layer Chickens to Vaccination with Commercial F Strain Mycoplasma gallisepticum Vaccine

A Chronicle of Serologic Response in Commercial Layer Chickens to Vaccination with Commercial F Strain Mycoplasma gallisepticum Vaccine

A Chronicle of Serologic Response in Commercial Layer Chickens to Vaccination with Commercial F Strain Mycoplasma gallisepticum Vaccine

Vaccination of multi-age layer operations, wherein one million plus commercial layer chickens are housed, has been spurious until the development of a self-propelled, constant-speed spray vaccinator. Still, even with its use, live Mycoplasma gallisepticum (MG) vaccinations have been questionable in terms of seroconversion. Using the vaccinator as a research tool over the past 5 yr, factors have been elucidated which impact seroconversion to one live MG vaccine in particular, the F strain of MG (FMG). These factors include the type of nozzle used to spray the vaccine, the temperature of the water used to rehydrate and administer the vaccine, and the pH and osmolarity of the fluid used to apply the vaccine. In the present study, one farm was monitored for its seroconversion rates over 4 1/2 yr, during which time the FMG vaccination protocol was amended as factors were identified that enhanced seroconversion rates. The results of this study showed that implementation and inclusion of the optimized factors into the vaccination protocol for FMG enhanced seroconversion rates because they went from an initial 50%-55% positive seroconversion rate to a consistent 100% positive seroconversion rate over the 56-mo study period.

A safety assessment of a fowlpox-vectored Mycoplasma gallisepticum vaccine in chickens

A recombinant fowlpox virus vaccine expressing key protective Mycoplasma gallisepticum antigens could facilitate in the prevention both of fowlpox virus and M. gallisepticum infections. Vectormune FP-MG vaccine, a recombinant fowlpox virus expressing both M. gallisepticum 40k and mgc genes, was assessed for its safety in 8-wk-old specific-pathogen-free White Leghorn chickens. The vaccine virus was serially passaged 5 times by wing-web inoculation. Based on the postinoculation clinical observation, gross pathological examination of air sacs and peritoneum, genetic stability evaluation, virus shedding and tissue distribution detection, horizontal transmission ability determination, and protection against fowlpox virus challenge, the Vectormune FP-MG vaccine possesses a high level of safety.

Influence of supplemental dietary poultry fat on the yolk characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

Effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5% supplemental dietary poultry fat (PF) on the egg yolk characteristics of commercial layers at 24, 34, 44, 50, and 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 and 22 wk of age and dietary treatments (basal control and basal control with 1.5% supplemental PF) were initiated at 20 wk of age. Yolk lipid concentration was reduced on wk 24 in birds that had been inoculated at 12 or 22 wk of age with FMG. The use of 1.5% supplemental PF increased percentage of yolk weight and yolk:albumen ratio across age and inoculation treatment. At 58 wk of age, concentrations of yolk palmitic acid increased and those of oleic and linolenic acid decreased when sham inoculations were given at 22 rather than at 12 wk of age. However, FMG inoculations given at 22 rather than at 12 wk increased palmitoleic acid and decreased stearic acid yolk concentrations. At 12 wk of age, FMG inoculations decreased yolk palmitoleic, oleic, and linolenic acid concentrations while causing increased yolk stearic and arachidonic acid levels when compared with sham inoculations. Furthermore, 1.5% supplemental PF decreased concentrations of palmitic and oleic acid and increased those of linoleic acid in the yolk at 58 wk of age. Despite the interaction of 1.5% supplemental PF with the prelay inoculation of FMG on early (18 to 26 wk) layer performance noted in a previous report, the effects of a prelay FMG inoculation and 1.5% supplemental PF on the egg yolk characteristics examined in the current study were independent of each other. This suggests that 1.5% supplemental PF is not effective in modulating the effects of an FMG inoculation at 12 wk of age on hen egg yolk characteristics between 24 and 58 wk of age and that the combined effects of PF supplementation and FMG inoculation on performance do not influence egg yolk characteristics.

Effect of Dosage and Vaccination Route on Transmission of a Live Attenuated Mycoplasma gallisepticum Vaccine: A Broiler Model *

Effect of Dosage and Vaccination Route on Transmission of a Live Attenuated Mycoplasma gallisepticum Vaccine: A Broiler Model *

Effect of Dosage and Vaccination Route on Transmission of a Live Attenuated Mycoplasma gallisepticum Vaccine: A Broiler Model

Mycoplasma gallisepticum (MG) is an economically significant pathogen of poultry species. Among the table egg sector of the poultry industry, live attenuated strains of MG are commonly used to limit production losses associated with MG-induced disease. These vaccines, however, may be problematic to broiler- and turkey-related industries because of associated virulence; therefore, an understanding of the transmissibility of the live MG vaccines is of particular importance. In the present study, a broiler model addresses the effect of vaccine application route and dosage on the transmission of the MG vaccine FVAX-MG to commingled unvaccinated subjects for 7 wk postvaccination. Vaccinations occurred at 2 wk of age via eyedrop or spray application at 1 x (4 x 10(6) colony-forming units [cfu]), 10(-3) x (4 x 10(3) cfu), or 10(-6) x (4 cfu) of the manufacturer's recommended dosage, and subsequent transmission to unvaccinated subjects was measured. The serologic response to MG antigen and the presence of MG DNA indicated FVAX-MG transmission only within the 1 x FVAX-MG eyedrop treatment. Among no other treatment was transmission of FVAX-MG detected. The results of the present study demonstrate that the dosage and vaccination route may have direct implications on subsequent transmission of FVAX-MG.

Influence of supplemental dietary poultry fat on the performance of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

The effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5% supplemental dietary poultry fat (PF) on the performance of commercial layers between 20 and 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 (before lay) and 22 (early in lay) wk and dietary treatments (basal control diets and basal control diets with 1.5% supplemental PF) were initiated at 20 wk of age. Mortality at wk 47 and 53 was greatest in birds inoculated with FMG at 22 wk. Feed consumption from 20 to 23 and from 52 to 55 wk of age was greater in birds that were inoculated with FMG (12 or 22 wk). However, feed consumption decreased in birds that were inoculated at 12 wk (sham or FMG) when 1.5% supplemental PF was added to the diet. Percentage of total egg production (EP) between 22 and 58 wk of age was highest in hens that were inoculated with FMG at 22 wk. Furthermore, weekly EP increased at wk 27 and 58 and decreased at wk 47 after birds had been inoculated with FMG (12 or 22 wk), and increased at 22 wk and decreased at 54 wk when inoculations (sham or FMG) were given at 22 wk. Egg weight was increased at wk 29, 31, 39, 40, 42, 44, 53, and 58 in birds that were inoculated with FMG (12 or 22 wk); however, there were no coherent treatment effects on eggshell quality. An FMG inoculation at 22 wk may promote total EP through 58 wk, whereas the inoculation of commercial layers with FMG (12 or 22 wk) may increase subsequent feed consumption during the early and late stages of EP and increase egg weight throughout lay. However, the supplementation of hen diets with 1.5% PF beginning at 20 wk of age may reduce subsequent feed consumption throughout lay in birds having experienced a prelay (12 wk) inoculation.

Effects of time-specific F-strain Mycoplasma gallisepticum inoculation overlays on prelay ts-11-strain M. gallisepticum vaccination on digestive and reproductive organ characteristics of commercial egg-laying hens ,

Two trials were conducted to determine the effects of a prelay ts-11-strain Mycoplasma gallisepticum (ts-11MG) vaccination alone or in conjunction with F-strain M. gallisepticum (FMG) inoculation overlays at 2 different age periods during lay on the digestive and reproductive organ characteristics of commercial egg-laying hens. In each trial, the following 4 treatments were utilized: sham vaccination at 10 wk of age, ts-11MG vaccination at 10 wk of age, ts-11MG at 10 wk of age overlaid by FMG inoculation at 22 wk of age, and ts-11MG at 10 wk of age overlaid by FMG at 45 wk of age. Necropsies were performed at the end of both trials (58 wk of age), using 2 birds from each of 4 replicate units per treatment, to observe treatment effects on the following parameters: liver weight, liver lipid and moisture concentrations, incidence of fatty liver hemorrhagic syndrome, ovary weight, number of mature ovarian follicles, and the total and segmental weights, lengths, and histologies of the oviduct and small intestine. Treatments affected only vaginal length as a percentage of total oviduct length. Vaginas were relatively longer in hens that had only been vaccinated with ts-11MG at 10 wk in comparison to all the other treatment groups, including controls. Except for relative vaginal length, the digestive and reproductive organs of layers were not influenced by the ts-11MG and FMG treatment regimens imposed in this study. These results confirm that when coupled with FMG inoculations during lay, prelay ts-11MG vaccinations may be a practical substitute for prelay FMG inoculations for providing continual protection against field-strain M. gallisepticum infections in layers.

Effects of time-specific F-strain Mycoplasma gallisepticum inoculation overlays on prelay ts-11-strain M. gallisepticum vaccination on blood characteristics of commercial laying hens

Two trials were conducted to determine the effects of a prelay ts-11-strain Mycoplasma gallisepticum (ts-11MG) vaccination alone or in combination with subsequent time-specific F-strain M. gallisepticum (FMG) inoculations on the blood characteristics of commercial laying hens. The following 4 treatments were utilized: 1) sham vaccination at 10 wk of age, 2) vaccination of ts-11MG at 10 wk, 3) ts-11MG at 10 wk overlaid by FMG inoculation at 22 wk, and 4) ts-11MG at 10 wk overlaid by FMG at 45 wk. Parameters measured in both trials were whole blood hematocrit, plasma protein, serum cholesterol, serum triglycerides, and serum calcium. No significant age x treatment interactions and no significant age or treatment main effects were observed for any of the blood parameters investigated, except for serum calcium. At wk 22, serum calcium concentrations were increased by vaccination with ts-11MG at 10 wk, and levels were further increased when the ts-11MG vaccination at 10 wk was overlaid by an FMG inoculation at 22 wk. These results suggest that ts-11MG vaccination at 10 wk of age alone or combined with F-strain inoculum overlays at either 22 or 45 wk may be used without any consequential effects on hematocrit or the lipid and protein levels in the blood of commercial layers. Because elevations in serum calcium were not associated with changes in hen performance, as reported in a previous companion article, it is further suggested that prelay ts-11MG vaccination before FMG inoculation overlays during lay may provide adequate protection against field strain M. gallisepticum infections while being innocuous to layer performance.

Effects of 6/85-strain Mycoplasma gallisepticum vaccination alone at ten weeks of age or in conjunction with F-strain Mycoplasma gallisepticum inoculation overlays at twenty-two or forty-five weeks of age on the reproductive and digestive organs of commercial egg-laying hens

Two trials were conducted to determine the effects of a prelay 6/85-strain Mycoplasma gallisepticum (6/85MG) vaccination alone or in conjunction with time-specific F-strain M. gallisepticum (FMG) inoculation overlays on the gross reproductive and digestive organ characteristics of commercial egg-laying hens. In each trial, the following 4 treatments were applied: 1) sham vaccination at 10 wk of age; 2) vaccination of 6/85MG at 10 wk; 3) 6/85MG at 10 wk overlaid by FMG inoculation at 22 wk; and 4) 6/85MG at 10 wk overlaid by FMG at 45 wk. Two birds per isolation pen (experimental replicate unit) were necropsied at the end of both trials to observe the effects of treatment on liver weight, liver lipid and moisture concentrations, incidence of fatty liver hemorrhagic syndrome, ovary weight, mature ovarian follicle numbers, and the total and segmental weights, lengths, and histologies of the oviduct and small intestine. The applied treatments affected only liver moisture. Liver moisture content was greater in birds vaccinated with 6/85MG at 10 wk alone or in conjunction with FMG at 45 wk in comparison with sham vaccinated controls and birds that received a 6/85MG vaccination at 10 wk overlaid by an FMG inoculation at 22 wk. Prelay 6/85MG vaccinations may be a suitable substitute for prelay FMG inoculations, and FMG overlays during lay on prelay 6/85MG vaccinations may also provide continual protection against field-strain MG infections without eliciting any subsequent suppressive effects on performance, as noted in an earlier study.

Effects of Prelay 6/85-Strain Mycoplasma gallisepticum Inoculation Alone or in Conjunction with the Inoculation of F-Strain Mycoplasma gallisepticum During Lay on the Blood Characteristics of Commercial Egg-Laying Hens

The effects of 6/85-strain Mycoplasma gallisepticum (6/85MG) inoculation alone or in conjunction with F-strain M. gallisepticum (FMG) overlays and their timing on the blood characteristics of commercial egg-laying hens were investigated. Control birds received sham inoculations at 10 wk of age. Birds in a second treatment group were inoculated with 6/85MG at 10 wk of age, those in a third treatment group were inoculated with 6/85MG at 10 wk followed by an overlay inoculation of FMG at 22 wk, and those in a fourth treatment group were inoculated with 6/85MG at 10 wk followed by an overlay inoculation of FMG at 45 wk. Parameters investigated at 24, 32, 43, and 47 wk were hematocrit, plasma total protein, and serum calcium, triglycerides, and cholesterol. No significant treatment effects were noted for hematocrit, serum triglycerides, or serum cholesterol. However, at wk 32, plasma protein was greater in birds that received 6/85MG at 10 wk or 6/85MG at 10 wk and FMG at 22 wk in comparison to controls. Also, at wk 47, serum calcium concentration was greater in birds that received 6/85MG at 10 wk and FMG at 45 wk compared with controls and those that received 6/85MG at 10 wk and FMG at 22 wk. These results suggest that the prelay inoculation of pullets with 6/85MG may subsequently elevate plasma protein, and in conjunction with an FMG overlay at 45 wk, may increase serum calcium concentrations in laying hens.

Differentiation of Mycoplasma gallisepticum Vaccine Strains TS-11 and 6/85 from Commonly Used Mycoplasma gallisepticum Challenge Strains by PCR*

Differentiation of Mycoplasma gallisepticum Vaccine Strains TS-11 and 6/85 from Commonly Used Mycoplasma gallisepticum Challenge Strains by PCR*

Differentiation of Mycoplasma gallisepticum Vaccine Strains ts-11 and 6/85 from Commonly Used Mycoplasma gallisepticum Challenge Strains by PCR

Mycoplasma gallisepticum (MG) is an important avian pathogen causing significant economic losses within the poultry industry. In an effort to develop tools to aid in MG research and diagnostics, we have compared sequences of the attenuated MG vaccine strain ts-11 to those of commonly used pathogenic challenge strains in search of a simple means of differentiation. Via gapA sequence alignments and comparisons, we have identified and designed primers facilitating strain differentiation. When applied to conventional polymerase chain reaction (PCR) assay at low annealing temperature, the primer sets allow for the differentiation of MG attenuated vaccine strains ts-11 as well as the attenuated MG vaccine strain 6/85 from the commonly utilized MG challenge strains R(low), R, and S6. Conventional PCR differentiation is based on the visualization of sole products with the attenuated MG strains ts-11 and 6/85 and the lack of the corresponding products from MG strains R(low), R, and S6. When applied to MG strain F, product visualization varies with the applied primer set. The differentiation of MG strains ts-11 and 6/85 from the pathogenic challenge strains was also accomplished via real-time analyses, however, the primer sets were not able to differentiate MG strains ts-11 and 6/85 from selected MG field isolates.

Development and immunogenicity of recombinant Mycoplasma gallisepticum vaccine strain ts-11 expressing chicken IFN-γ

Mycoplasma gallisepticum (MG) is a poultry pathogen that causes respiratory disease and loss of egg production worldwide. A live attenuated vaccine, ts-11, has been used for the control of MG in several countries. To improve the functionality of the vaccine and investigate its potential as a delivery vector for host immune molecules and foreign antigens, we have developed ts-11 as a vector to express and secrete chicken IFN-γ (ts-11 C3) using a transposon-based delivery vector. Following administration of ts-11 C3 in chickens by eye drop, up to 2 weeks post-vaccination, neither significant systemic IFN-γ expression nor an antibody response as determined by the rapid serum agglutination (RSA) could be detected, while moderate RSA scores were detected in birds vaccinated with ts-11. However, the MG-specific IFN-γ response in spleen cultures was significantly enhanced in ts-11 C3 vaccinated chickens and, more interestingly, significant heterophil infiltration was detected in the tracheal epithelium in ts-11 C3 vaccinated birds, but not in ts-11 vaccinated birds. These results indicate that the IFN-γ expressed by ts-11 C3 enhanced host cellular immunity rather than humoral immunity and may also have stimulated mucosal heterophil infiltration. These results also suggest that ts-11 is a promising vector for protective antigens of other chicken respiratory pathogens.