Effects of the in ovo injection of an Escherichia coli vaccine on the hatchability and quality characteristics of commercial layer hatchlings.

In the commercial egg industry, avian pathogenic Escherichia coli (APEC) can lead to significant economic loss. The Poulvac E. coli vaccine (PECV) is a commercially available attenuated live vaccine commonly applied via spray or drinking water to protect against losses associated with colibacillosis. The PECV has not been tested in layer hatching eggs using in ovo injection. Therefore, the purpose of this experiment was to determine the effects of injecting 50 µL of different doses of the PECV into Hy-Line W-36-layer hatching eggs on the hatchability and quality characteristics of hatchlings. At 18 d of incubation (DOI), treatments included 1 noninjected and 1 diluent-injected control. Furthermore, PECV treatments included a full dose (4.4 × 108E. coli CFU) or serial dilutions of the full dose to produce 4.4 × 106, 4.4 × 104, or 4.4 × 102 CFU doses of E. coli. In ovo injections targeted the amnion. Percent hatchability of live embryonated eggs (HI), percent residue eggs, hatchling mortality, and female chick whole and yolk-free BW, relative yolk sac weight, and body length were among the variables examined. Treatment significantly (P < 0.0001) affected HI, with HI being highest in the control groups (97.3% in the noninjected and 94.2% in the diluent-injected), and with HI values being 89.0, 88.9, 84.4, and 71.2% in the 4.4 × 102, 4.4 × 104, 4.4 × 106, and 4.4 × 108 CFU E. coli dose treatments, respectively. The percentage of live embryos that did not complete hatch but that pipped internally (P = 0.024) or externally (P < 0.0001) were significantly affected by treatment, with percentages being highest in the 4.4 × 108 CFU treatment. Female chick body length was significantly (P < 0.0001) affected by treatment and was longer in both control groups and in the 1 × 102 CFU E. coli treatment in comparison to all other treatments. Yolk-free female chick BW was significantly (P = 0.034) affected by treatment and was lower in the 4.4 × 106 CFU and 4.4 × 108 CFU treatments when compared to the diluent-injected control group. An increase in the E. coli concentration administered in the amnion of embryonated layer hatching eggs at 18 DOI decreased hatch success and female chick yolk-free BW and body length.

Effects of dietary Original XPC on selected blood variables in layer pullets challenged with Mycoplasma gallisepticum,.

Effects of dietary Original XPC (XPC) on 17 selected blood variables in commercial layer pullets challenged with the virulent, low-passage R strain of Mycoplasma gallisepticum (RlowMG) were investigated. Hy-Line W-36 pullets sourced from M. gallisepticum-clean layer breeders were fed a basal diet with XPC (1.25 kg/metric ton) or without from hatch until 12 wk of age (woa). At 8 and 10 woa, half of the birds in each dietary treatment were challenged with RlowMG. Blood samples were taken immediately before the initial RlowMG challenge at 8 woa and again at 12 woa (4 wk after challenge). At 8 woa, blood pH was lower and glucose concentration was higher in the preassigned challenge treatment groups. At 12 woa, the concentration of oxygen dissolved in the blood was significantly lower in the RlowMG-challenged group than the unchallenged group of birds regardless of dietary treatment. The RlowMG challenge significantly increased blood carbon dioxide partial pressure, calcium, sodium, anion gap, osmolality, glucose, and corticosterone levels but significantly decreased blood oxygen partial pressure, oxyhemoglobin concentration, concentration of oxygen dissolved in the blood, chloride, and pH levels. Because blood pH and glucose concentration at 8 woa were examined before challenge, their baseline values were biased with respect to challenge treatment before treatment was applied. However, the lack of a significant main effect due to diet at 8 woa for blood pH and glucose concentration, along with the other 15 blood variables, indicate that the baseline data with respect to dietary treatment were unbiased, allowing for real dietary effects to be accurately assessed. In conclusion, layer pullets challenged with RlowMG undergo a stress response associated with changes in various physiological blood variables, and a decrease in pH and increase in carbon dioxide partial pressure, in association with a lack of change in bicarbonate, indicates that the stress response caused by the RlowMG challenge was associated with respiratory acidosis. Nevertheless, feeding XPC did not influence the effects of challenge treatment on these postchallenge physiological blood values.

Growth and humoral immune effects of dietary Original XPC in layer pullets challenged with Mycoplasma gallisepticum.

Effects of dietary Original XPC (XPC) in commercial layer pullets challenged with the virulent, low passage R strain of Mycoplasma gallisepticum (Rlow MG) were investigated. Hy-Line W-36 pullets sourced from MG-clean breeders were fed a basal diet with or without (CON) XPC (1.25 kg/metric ton) from hatch until 12 wk of age (woa). At 8 and 10 woa, half of the birds in each dietary treatment were challenged with Rlow MG. Body weight was recorded at 3, 8, and 12 woa, and ovary, ceca, and bursa weights were recorded at 3 and 12 woa. Blood samples were taken immediately before the initial Rlow MG challenge at 8 woa and again at 12 woa to test for IgM and IgG antibody production against MG. All birds were evaluated for MG lesion scores at 12 woa. Regardless of challenge, inclusion of XPC in the diet did not significantly alter BW at 3 or 8 woa or relative organ weights at 3 or 12 woa. However, at 12 woa, BW of XPC-fed birds, regardless of challenge was significantly (P = 0.0038) heavier than CON by 25.7 g. All birds tested negative for MG antibodies before the 8 woa challenge. Respective percentage serum plate agglutination and ELISA positive birds at 12 woa were 0 and 0% (CON, nonchallenged), 1.4 and 0% (XPC, nonchallenged), 100 and 47.2% (CON, challenged), and 100 and 50.0% (XPC, challenged). Diet did not significantly affect ELISA titers, but they were significantly (P < 0.0001) increased due to challenge. Furthermore, lesion scores were significantly higher for Rlow MG-challenged birds (P = 0.0012), and dietary treatment with XPC in challenged birds numerically reduced MG lesion scores from 0.278 to 0.194. In conclusion, although dietary XPC did not significantly alter the humoral immune response, antibody titer levels, or severity of MG lesions in layer pullets that were or were not challenged with Rlow MG, it led to an increase in their rate of growth through 12 woa.

Effects of different vaccine combinations against Mycoplasma gallisepticum on the digestive and reproductive organ characteristics of commercial egg-laying hens.

Mycoplasma gallisepticum (MG) is a major and economically significant pathogen of avian species. When administered before lay, F-strain MG (FMG) can reduce egg production during lay, but the ts-11 strain of MG (ts11MG) does not exert this effect. Two trials were conducted to determine the effects of pre-lay vaccinations of ts11MG, MG-Bacterin (MGBac), or their combination, in conjunction with an FMG vaccination overlay after peak production on the digestive and reproductive organ characteristics of Hy-Line W-36 layers housed in biological isolation units (4 units per treatment, 10 birds per unit). The following vaccination treatments were administered at 10 wk of age (woa): 1) Control (no vaccinations); 2) MGBac; 3) ts11MG; and 4) ts11MG and MGBac combination (ts11MG+MGBac). At 45 woa, half of the birds were vaccinated with a laboratory stock of high passage FMG. In both trials, parameters determined in 4 birds per unit at 55 woa included: BW; fatty liver hemorrhagic syndrome incidence; mean number of mature ovarian follicles; ovarian, oviduct, and small intestine weights; and the weights and lengths of the various portions of the oviduct and small intestine. Treatment effects were observed for the weights of the entire small intestine and the duodenum, jejunum, and ileum, as percentages of BW; and for vagina weight as a percentage of total oviduct weight. In general, the weights of the small intestine and its 3 components were increased in response to the FMG vaccine that was administered at 45 woa. An FMG vaccination at 45 woa may increase relative intestine weight in layers; however, use of a prelay MGBac vaccine alone or in combination with ts11MG, with or without an FMG overlay, does not affect the gross characteristics of their digestive and reproductive organs, and may be used without having an adverse effect on their performance, as was observed in a previous companion study.

The impact of deposition site on vaccination efficiency of a live bacterial poultry vaccine.

Vaccines are utilized within the poultry industry to minimize disease-associated losses and spray vaccination is a commonly utilized means for the mass application of poultry vaccines. During this process, vaccine-laden particles are deposited upon target areas (e.g., eyes, nares, and oral cavity) resulting in the direct internalization of the vaccine. However, particles are also deposited on nontarget areas such as the exterior of the subject and its surrounding environment. To better determine the fate of particles deposited upon nontarget areas and the impact of deposition site on the efficiency of vaccine application, a live bacterial poultry vaccine (AviPro(®) MG F) was applied via spray using a spray cabinet with a slotted partition allowing for head-only, body-only, and whole-bird spray application. At 11 wk age, Hy-Line(®) W-36 pullets (n = 280) were allocated equally among 7 treatments including: nonvaccinated controls, pullets spray-vaccinated at the manufacturer's recommended dose (1X) in a site-specific manner (head-only, body-only, and whole-bird), pullets spray-vaccinated at 5X the recommended level (body-only), pullets vaccinated by manual eye-drop application (1X), and pullets eye-drop vaccinated at a level approximating that achieved during the spray vaccination process (1/700X). At 6 to 7 wk postvaccination, vaccination efficiency was assessed via serological-based assays [serum plate agglutination (SPA) and ELISA] and the detection of vaccine-derived in vivo populations. Results indicate an additive contribution of the vaccine deposited on the body to the overall vaccination efficiency of this live bacterial live poultry vaccine.

Effects of different vaccine combinations against Mycoplasma gallisepticum on the internal egg and eggshell characteristics of commercial layer chickens 1,2,3.

Live F-strain Mycoplasma gallisepticum (FMG) vaccines are presently being used to help control field-strain MG outbreaks. However, they may exert some adverse effects on egg production. Live strains of MG of lesser virulence as well as killed vaccines have little or no effect on egg production, but afford lower levels of protection. This has led to research investigating their use in combination with a subsequent overlay vaccination of FMG given later in the production cycle. In the present study, 2 trials were conducted to investigate the effects of prelay vaccinations of live and killed MG vaccines or their combination, in conjunction with an FMG vaccine overlay after peak production, on the egg characteristics of commercial layers. The following vaccination treatments were administered at 10 wk of age (woa): 1) unvaccinated (Control), 2) MG-Bacterin (MGBac) vaccine, 3) ts-11 strain MG (ts11MG) vaccine, and 4) MGBac and ts11MG combination (MGBac + ts11MG). At 45 woa, half of the birds were overlaid with an FMG vaccine. In each trial, internal egg and eggshell parameters including egg weight (EW), Haugh unit score (HU), eggshell breaking strength (EBS), percentage yolk weight (PYW), percentage albumen weight (PAW), percentage eggshell weight (PSW), eggshell weight per unit surface area (SWUSA), percentage yolk moisture (PYM), and percent total lipids (PYL) were determined at various time periods between 21 and 52 woa. At 28 woa, SWUSA was lower in the ts11MG and MGBac + ts11MG groups compared to the Control group. Conversely, at 43 woa, SWUSA was higher in the ts11MG than in the MGBac group. Between 23 and 43 woa, PYL was higher in the MGBac and ts11MG groups in comparison to the Control group. In conclusion, vaccination with MGBac alone or in combination with ts11MG at 10 woa with or without an FMG vaccine overlay at 45 woa does not adversely affect the internal egg or eggshell quality of commercial layers throughout lay.

Effects of live and killed vaccines against Mycoplasma gallisepticum on the performance characteristics of commercial layer chickens.

Different vaccine strains of Mycoplasma gallisepticum have been used on multiple-age commercial layer farms in an effort to protect birds against virulent field-strain infections. Use of the F-strain of M. gallisepticum (FMG), as an overlay vaccine during lay, may be necessary because of the lower level of protection afforded by M. gallisepticum vaccines of low virulence given before lay. Two replicate trials were conducted to investigate effects of live and killed M. gallisepticum vaccines administered individually and in combination before lay, in conjunction with an FMG vaccine overlay after peak egg production (EP), on the performance characteristics of commercial layers. The following treatments were utilized at 10 wk of age (woa): 1) control (no vaccinations); 2) ts11 strain M. gallisepticum (ts11MG) vaccine; 3) M. gallisepticum-Bacterin vaccine (MG-Bacterin); and 4) ts11MG and MG-Bacterin vaccines combination. At 45 woa, half of the birds were overlaid with an FMG vaccine. Hen mortality, BW, egg weight, percentage hen-day EP, egg blood spots, and egg meat spots were determined at various time periods between 18 and 52 woa. The data from each trial were pooled. Treatment did not affect performance in interval I (23 to 45 woa). However, during interval II (46 to 52 woa), the EP of control and MG-Bacterin-vaccinated birds that later received an FMG vaccine overlay was lower than that in the other treatment groups. Furthermore, treatment application reduced bird BW during interval II. Despite the effects on BW and EP, no differences were observed for egg blood or meat spots among the various treatments. It is suggested that the vaccination of commercial layers before lay with ts11MG, but not MG-Bacterin, may reduce the negative impacts of an FMG overlay vaccination given during lay. These results establish that the vaccination of pullets with ts11MG in combination with the vaccination of hens with an FMG overlay, for continual protection against field-strain M. gallisepticum infections, may be used without suppressing performance.

Impact of fowlpox-vectored Mycoplasma gallisepticum vaccine Vectormune FP MG on layer hen egg production and egg quality parameters.

This study was conducted to determine the impact of vaccination with Vectormune FP MG on egg production and egg quality characteristics of Single Comb White Leghorn hens. Due to questions of the efficacy of this vaccine in preventing Mycoplasma gallisepticum-mediated pathology, the ability of this vaccine to protect against postproduction-peak egg losses associated with F-strain M. gallisepticum (FMG) vaccination was also investigated. Vaccination with Vectormune FP MG did not result in any significant change in egg production or egg quality parameters compared with control (unvaccinated) hens. Subsequent revaccination with FMG at 45 wk of age (woa) yielded no impact on egg production or egg quality parameters of Vectormune FP MG vaccinated hens, unlike prior results for postproduction-peak vaccination of M. gallisepticum-clean hens with FMG, which exhibited a drop in egg production of approximately 6%. No difference in egg size distribution was observed for any of the treatment groups before or after FMG revaccination. These results suggest that hens can be safely vaccinated with Vectormune FP MG as pullets and can be revaccinated with a live M. gallisepticum vaccine such as FMG at a later date with no deleterious effects on egg production or egg or eggshell quality parameters.

Mycoplasma gallisepticum transmission: comparison of commercial F-strain vaccine versus layer complex-derived field strains in a tunnel ventilated house.

Two simultaneous trials were conducted using a commercially available, live, F strain Mycoplasma gallisepticum (FMG) vaccine (trial 1) or 2 inocula of layer complex-derived MG strains (LCD-MG; trial 2). In each of the 2 trials, 4 commercial turkeys were housed in each of 2 adjoining pens immediately adjacent to air inlets. The turkeys (8/trial) were inoculated in the right eye with either a 1× dose of FMG (trial 1) or with 0.02 mL of 1 of 2 actively growing LCD-MG inocula (4 turkeys/inoculum; trial 2). In each of the 2 trials, one pen housing 4 inoculated turkeys was maintained without the addition of other poultry, whereas 16 MG-free broilers and 4 MG-free layers were added to the other pen of 4 inoculated turkeys. Within each of the trials and at increasing intervals, either 4 layers (3 pens) or 4 turkeys (3 pens) were placed down-airstream from the inoculated pens. The distance of the first pen from the inoculated turkeys was separated by the width of one pen that was empty. Succeeding down-airstream pens were situated such that the empty distance (absence of any poultry) between pens that contained poultry doubled from one pen to the next such that the final pen that contained poultry had 4 empty pens between it and the next up-airstream pen that also contained poultry. At 106 d postinoculation, all poultry were bled, swabbed for MG from the choanal cleft, and then euthanized and necropsied. No commingled poultry in trial 1 (FMG), whether inoculated (turkeys) or commingled (layers and broilers), died during the course of the trial, and 5 of the 8 FMG-vaccinated turkeys exhibited serological but not cultural evidence of mycoplasmosis. In trial 2 (LCD-MG), 2 commingled broilers died and no inoculated turkeys exhibited either serological or cultural evidence of mycoplasmosis. In both trials, no poultry housed down-airstream from the inoculated poultry showed evidence of clinical signs of mycocplasmosis and none showed either serological or cultural evidence of mycoplasmosis.

Effect of infection route and concurrent infectious bronchitis virus vaccination on Mycoplasma gallisepticum disease pathology in an experimental model.

The study of Mycoplasma gallisepticum (MG) infection is needed, not only to understand the disease process but also to understand the mechanisms by which MG vaccines protect the host. Many model systems have been used to study the MG disease process. This work compared two different routes of infection (intratracheal versus eye drop) in commercial pullets, looking for differences in the pathology (air sac and tracheal lesion scores, and tracheal mucosal thickness) and the humoral immune response (measured by serum plate agglutination) of the host. The impact of concurrent infectious bronchitis virus vaccination on disease outcomes was also determined. Results showed that the intratracheal route of MG infection caused increased air sac and tracheal lesion scores and tracheal mucosal thickness at one week post infection, whereas the eye drop route produced no noticeable pathology. However, tracheal mucosal thicknesses of intratracheally challenged pullets were not statistically different from those of the eye drop challenged or control pullets at two and three weeks post infection. Concurrent infectious bronchitis virus vaccination had a negligible outcome on disease pathology. Vaccination of specific-pathogen-free chickens with the F-strain MG vaccine completely protected them against the effects of MG intratracheal infectious challenge, as evidenced by a lack of significant difference in air sac and tracheal lesion scores and tracheal mucosal thickness with those of unchallenged media control chickens.

A comparative study of live attenuated F strain-derived Mycoplasma gallisepticum vaccines.

Commercially available attenuated strains of Mycoplasma gallisepticum (MG) are commonly used within the layer industry to control MG-induced mycoplasmosis. Among these are two live MG vaccines derived from the moderately pathogenic MG "chick F" strain. In the present study, the commercially available F strain derivatives were compared for their ability to elicit seroconversion, persist in vivo, and protect against virulent MG-induced airsacculitis. In addition, a noncommercial laboratory-derived high-passage F strain isolate was included in the study. Commercial (Hy-Line W-36) layers were placed in biological isolation units at 9 wk of age (woa). At 10 woa, birds within each biological isolation unit were treated via eye-drop application with one of the three F strain-derived vaccines at one of four levels (1x, 10(-1)x, 10(-2)x, or 10(-3)x). For the commercially available F strain derivatives, 1x equaled the manufacturer's recommended dose. The 1x dose of the noncommercial laboratory-maintained F strain derivative equaled 20 microl of a 48 hr culture. For wk 1-6 postvaccination (p.v.), sera were collected weekly from each bird, and seroconversion was assessed via serum plate agglutination (SPA). Virulent MG (strain R(low)) challenge occurred via intratracheal inoculation at 7 wk p.v. Necropsies were subsequently performed to assess challenge-associated airsacculitus. For each F strain derivative applied at 1x and 10(-1)x, 100% seroconversion, as measured by SPA, was demonstrated by 6 wk p.v., and rates at the 10(-2)x dosage were 10% and 90% for the commercial vaccines and 60% for the laboratory-derived strain in this period. Following challenge, airsacculitis was observed in 66.67% of the nontreated controls but not in any 1x- or 10(-1)x-treated bird independent of applied F strain derivative.

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.

Effects of vaccination with F-strain Mycoplasma gallisepticum on egg production and quality parameters of commercial layer hens previously vaccinated with 6/85-strain Mycoplasma gallisepticum.

This study was conducted to determine the effect of overlaying (revaccinating) F-strain Mycoplasma gallisepticum at 22 or 45 wk of age on commercial leghorn hens previously vaccinated with 6/85-strain M. gallisepticum at 10 wk of age. The treatment groups included unvaccinated hens (group 1), hens receiving 6/85-strain M. gallisepticum only (group 2), and hens receiving 6/85-strain M. gallisepticum followed by F-strain M. gallisepticum at either 22 (group 3) or 45 (group 4) wk of age. There was no significant effect on egg production or egg size distribution between any of the treatment groups, unlike previous studies looking at F-strain vaccination only. Egg quality parameters, including eggshell strength, Haugh unit score, and blood-meat spot were similar between the different treatment groups. There was a difference in the rate of pimpling at postpeak production for the treatment group receiving F-strain M. gallisepticum at 22 wk of age, consistent with previously published results. This work suggests that hens previously vaccinated with 6/85-strain M. gallisepticum can be safely revaccinated with F-strain M. gallisepticum to increase protection from field strains while ameliorating the adverse effects associated with F-strain M. gallisepticum vaccination in layers post onset of lay.

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.

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.

Effect of selected water temperatures used in Mycoplasma gallisepticum vaccine reconstitution on titer at selected time intervals.

Numerous methods are currently used throughout the poultry industry for the administration of vaccines. Each utilizes water for vaccine reconstitution and/or administration, including two of the three commercially available live Mycoplasma gallisepticum (MG) vaccines. Selected water temperatures were used to reconstitute and/or dilute the three commercially available live MG vaccines. Water temperatures included 4 C, 22 C (room temperature), and 32 C, and titer (color change units) was recorded at four time intervals, at point of reconstitution (time 0), 15, 30, and 60 min postreconstitution of the vaccines (time periods 15, 30, and 60, respectively). Results for F strain MG (FMG) vaccine showed significant decreases in titer from time 0 to time 15 for the 22 C and 32 C water temperatures but no significant decrease for any time period for FMG reconstituted with 4 C water. For 6/85 strain MG no significant difference in titer was noted for any of four time periods within any of the three water temperatures. For ts-11 strain MG a significant decrease was observed in titer at each of the four postdilution time periods when diluted with 32 C water. There was no significant decrease in titer at any time period for ts-11 MG vaccine when diluted with either 4 C or 22 C water.

The effects of increasing sodium chloride concentration on Mycoplasma gallisepticum vaccine survival in solution.

Lyophilized Mycoplasma gallisepticum (MG) vaccines are generally rehydrated and diluted with distilled or chlorine-free water as per manufacturer recommendations. However, as mycoplasma species lack a cell wall, this can lead to decreased viability of live vaccine during administration. The ability of phosphate-buffered saline (PBS) to prevent losses in live vaccine viability was examined. It was shown that a concentration of 1 x PBS prevented the two-fourfold decrease in MG viability seen when the vaccines were diluted with water alone.

Effects of increased dosages of the Mycoplasma gallisepticum vaccine MYCOVAC-L in layer chickens subsequently challenged with virulent M. gallisepticum: egg production and serologic response.

Ten-week-old Hy-Line Commercial W-36 pullets were spray-vaccinated with MYCOVAC-L at the manufacturer's recommended dosage (1x) or at 15 times that rate (15x). At 22 or 45 wk of age, subsets of 1x- and 15x-vaccinated pullets were challenged with the virulent Mycoplasma gallisepticum (MG) strain Rlow. Percent hen-day egg production was determined through week 55. Analyses for treatment effects on overall (22-56 wk) percent hen-day egg production revealed no significant differences between nonchallenged 1x and nonchallenged 15x MYCOVAC-L treatments. Among 1x MYCOVAC-L-vaccinated groups, Rlow challenge at 45 wk corresponded to significantly (P < or = 0.01) lower overall egg production compared with the unchallenged 1x-vaccinated control (70.88% vs. 79.15%, respectively). Conversely, at the 15x MYCOVAC-L dosage level, overall egg production was not significantly affected by virulent MG challenge at 45 wk compared with its unchallenged counterpart (84.09% vs. 81.03%, respectively) and could indicate increased protection from virulent MG challenge. Serologic monitoring indicated the virulent MG challenge was consistently (100%) associated with seroconversion. Comparisons among the nonchallenged experimental treatments indicated that vaccinations at the 15x MYCOVAC-L dosage rate were associated with a greater seroconversion rate at weeks 21, 27, and 44, but not at week 50.

Unexpected results from the application of signature-tagged mutagenesis to identify Yersinia pestis genes required for adherence and invasion.

The signature-tagged mutagenesis (STM) method was applied in a protocol designed to identify genes required for Yersinia pestis invasion into epithelial cells. A library of 3060 mutants of Y. pestis CO99-3015 was made and assayed using an in vitro invasion assay with gentamicin protection. Initial results from the screen identified a set of 23 genes that might be required for invasion; however, screening of individual mutants for decreased invasion, even in a competition assay with the parent strain, failed to reveal obvious invasion defects. Altered colony character or size might have imposed a growth disadvantage for two of the mutants, which could possibly account for apparently decreased invasion. The sensitivity of the mutants to gentamicin was assayed to determine if the presence of the kanamycin-resistance cassette in the STM transposon changed the gentamicin resistance of the individual mutants. It was discovered that the mutants exhibited a variable range of resistance to killing by gentamicin, suggesting that the presence of the kanamycin-resistance cassette or the particular insertion mutation did in many cases affect the bactericidal potency of gentamicin. However, all mutants remained highly sensitive to growth inhibition in a disk assay on plates. These results may warrant precautions for use of kanamycin-resistance markers in studies with fully virulent Y. pestis, since gentamicin has been recommended for treatment of plague. Further, to use STM in the context of invasion assays, a selection other than gentamicin should be applied.

The long-term cancer survivor: a challenge for nurse practitioners.

For approximately 8 million Americans alive today, the words "cancer" and "survival" are no longer mutually exclusive. As advances are made in the early detection and treatment of cancer, the numbers of survivors who are recovering from their illnesses or living longer with cancer as a chronic disease are increasing. With this extended survival comes a new set of responsibilities and standards for follow-up care that include the following: (1) the recognition of chronic or potential problems, (2) the need for life-long surveillance, and (3) continued access to quality health care. Long-term follow-up care for cancer survivors can be a specialty unto itself. The development of clinics or programs that specialize in caring for this expanding population have a history within pediatric oncology. The challenge in the current health care market is to dedicate energy and resources to do the same within the adult oncology community. Nurse practitioners are ideal candidates to create holistic programs that focus on the special needs, both biomedical and psychosocial, of long-term cancer survivors.