Effects of probiotics and maternal vaccination on Salmonella enteritidis infection in broiler chicks

The effects of probiotics and maternal vaccination with an inactivated Salmonella Enteritidis (SE) vaccine on day-old chicks challenged with SE were evaluated. A 2 X 3 factorial arrangement was used (with or without probiotics; breeders nonvaccinated, vaccinated intramuscularly, or vaccinated intraperitoneally). Three trials were conducted in isolation cabinets and SE challenge was different between trials. The number of SE organisms per chick and the time interval between housing and introduction of seeder birds (hereafter called challenge) were 1.6 X 10(8) and 1 hr (Trial I), 1.8 X 10(6) and 12 hr (Trial II), and 1.2 X 10(4) and 24 hr (Trial III). SE recovery was assessed in ceca and liver at 3, 5, and 7 days postchallenge, and the number of colony-forming units (CFU) in ceca was evaluated at 5 and 7 days postchallenge. The number of SE (log CFU) in the ceca reduced 0.56 log (from 7.59 to 7.03) and 1.45 log (7.62 to 6.17) because of the treatment with probiotics in Trials II and III, respectively. The greater reduction in Trial III indicates the importance of the early use of probiotics on the prevention of SE infection. Treatment with probiotics resulted in a smaller number of SE-positive livers after 5 days postchallenge on Trial III. Although there was no significant effect of maternal vaccination on the number of SE CFU in the ceca, a significant effect of maternal vaccination on the SE CFU was observed in the liver, but not in the ceca at 5 days after challenge.     

Colonization of a marker and field strain of Salmonella enteritidis and a marker strain of Salmonella typhimurium in vancomycin-pretreated and nonpretreated laying hens

This study was conducted to evaluate the influence of a vancomycin pretreatment on the ability of marker (nalidixic-acid resistant) Salmonella Enteritidis (SE(M)), field Salmonella Enteritidis (SE(E)), and marker Salmonella Typhimurium (ST(M)) strains to colonize within the intestinal and reproductive tracts and translocate to other organs of leghorn laying hens. In each of three trials, caged laying hens (76, 26, and 33 wk ofage) were divided into six groups designated to receive SE(M), SE(F), or ST(M), and half were pretreated with vancomycin (n = 11-12 hens). Vancomycin-treated hens received 10 mg vancomycin in saline/kilogram body weight orally for 5 days to inhibit Gram-positive bacteria within the intestines. On Day 6, all hens were concurrently challenged by oral, intravaginal, and intracolonal routes with Salmonella and placed into separate floor chambers by Salmonella strain. Two weeks postinoculation, all hens were euthanatized and the ceca, spleen, liver/gall bladder (LGB), upper (URT), and lower (LRT) reproductive tracts, and ovarian follicles were aseptically collected, and analyzed for Salmonella. Results did not differ for the three hen's ages and were therefore combined. The vancomycin pretreatment also had no significant effect on the colonization ability of SE(M), SE(F) or ST(M), and therefore results were combined within Salmonella strain. The marker strain of Salmonella Enteritidis was recovered from 21% of ceca, 4% of LGB, 9% of LRT, and 17% of the fecal samples. The field strain of Salmonella Enteritidis was recovered from 88% of ceca, 96% of spleen, 92% of LGB, 30% of LRT, 4% of URT, 13% of follicle, and 42% of the fecal samples. The marker strain of Salmonella Typhimurium was recovered from 100% of ceca, 74% of spleen, 91% of LGB, 30% of LRT, 9% of URT, 9% of follicle, and 100% of the fecal samples. Among ceca, spleen, LGB, and fecal samples, SE(F) and ST(M) colonization was significantly greater than SE(M) colonization. Overall prevalence of Salmonella in the reproductive tracts of challenged hens was relatively low, ranging from 4% to 30%.     

Coarse-spray immunization of one-day-old broilers against enteric reovirus infections.

Coarse-spray (CS) administration of a commercial S1133 reovirus vaccine was evaluated in 1-day-old specific-pathogen-free broilers for prevention of clinical infection induced by intratracheal challenge with two enteric reovirus isolates. In Expt. 1, chickens were challenged at 4 days of age with either the 2408 or CO8 isolate. In Expt. 2, chickens were challenged at 7 days of age with either isolate. In Expt. 3, chickens were challenged at 3, 5, or 7 days of age with the 2408 isolate. In Expt. 1, vaccinated birds showed significant protection against challenge with either isolate at 4 days of age as measured by morbidity, mortality, gross lesions, and body weight. In Expt. 2, vaccinated birds showed greater protection against challenge at 7 days of age. In Expt. 3, resistance in vaccinated birds increased with time between vaccination and challenge. Vaccinated birds challenged at 3 days of age showed no significant protection, whereas vaccinated birds challenged at 5 or 7 days of age had increased resistance. This vaccine did not induce a drop in weight gain, morbidity, mortality, or microscopic lesions in the tendons.     

Use of bacteriophages in combination with competitive exclusion to reduce Salmonella from infected chickens

Salmonella-spedfic bacteriophages (BP) and competitive exclusion (CE) were used to reduce Salmonella colonization in experimentally infected chickens. A "cocktail" of distinct phage (i.e., phage showing different host ranges and inducing different types of plaques on Salmonella Typhimurium [ST] cultures) was developed. The killing activity of the selected BPs on ST cultures differed significantly, as determined in in vitro killing assays. BPs were administered orally to the chickens several days prior and after ST challenge but not simultaneously. BPs were readily isolated from the feces of the BP-treated chickens approximately 48 hr after administration. A CE product consisting of a defined culture of seven different microbial species was used either alone or in combination with BP treatment. CE was administered orally at hatch. Salmonella counts in intestine, ceca, and a pool of liver/spleen were evaluated in Salmonella-challenged chickens treated with BP or with BP and CE. In both trials 1 and 2, a beneficial effect of the phage treatment on weight gain performance was evident. A reduction in Salmonella counts was detected in cecum and ileum of BP-, CE-, and BP+CE-treated chickens as compared with nontreated birds. In trial 1, BP treatment reduced ST counts to marginal levels in the ileum and reduced counts sixfold in the ceca. A reduction of Salmonella counts with BP, CE, and BP+CE treatments was evident in chickens from trial 2. Both CE and BP treatments showed differences in the reduction of Salmonella counts after challenge between spedmens obtained at days 4 and 14 postchallenge in ceca, liver/spleen, and ileum. The preliminary data presented in this report show that isolation and characterization of Salmonella-specific BP is uncomplicated and feasible on a larger scale. Results indicate a protective effect of both Salmonella-specific BPs and a defined competitive exclusion product against Salmonella colonization of experimentally infected chickens. These results are encouraging for further work on the use of BP as an effective alternative to antibiotics to reduce Salmonella infections in poultry.     

Effect of a selected Lactobacillus spp.-based probiotic on Salmonella enterica serovar enteritidis-infected broiler chicks

The effect of a Lactobacillus spp.-based probiotic (FM-B11) on Salmonella enterica serovar Enteritidis (SE) recovery was evaluated in liquid (Expt. 1) and lyophilized (Expt. 2) forms in two separate experiments with two trials each. For each trial, 80 broiler chicks were randomly allocated into two treatments: control and probiotic culture. All chicks were challenged with SE (approximately 10(4) colony-forming units [cfu]) upon arrival at our laboratory. In both experiments, probiotic culture was administered in the drinking water for 3 consecutive days at a final concentration of approximately 10(6) cfu/ml, beginning 1 hr after SE challenge. Cecal tonsils were aseptically removed at 24 and 72 hr postchallenge, followed by enrichment and plating on xylose lactose deoxycholate (XLD) agar for the presence or absence of Salmonella-typical colonies. In Expt. 1, a significant reduction (P < 0.05) in SE-positive samples was observed in both trials at 24 and 72 hr postchallenge. Additionally, in Expt. 2, the lyophilized probiotic decreased (P < 0.05) SE recovery at both 24 and 72 hr postchallenge compared with the control group in trial 1. In trial 2, SE evaluation was performed only at 72 hr after challenge and fewer (P < 0.001) treated samples were positive for SE. Results showed that application of either liquid or lyophilized probiotic culture in the drinking water for 3 consecutive days can help to reduce SE recovery from young birds, although further research is needed to elucidate the mechanism of this response.     

In vivo biologic effects of recombinant-turkey interferon-gamma in neonatal leghorn chicks: protection against Salmonella enteritidis organ invasion.

Interferon-gamma (IFNgamma) has been demonstrated to have potent stimulatory effects on parameters of cell-mediated immunity in chickens (11). Protection of neonatal leghorn chickens against infection by invasive salmonellae has been associated with enhanced cell-mediated indices of immunity (5). The present investigation evaluated the effect of recombinant-turkey (rt) IFN-gamma on protection of neonatal leghorn chicks from Salmonella enteritidis (SE) organ invasion after experimental challenge in three experiments. In Expt. 1, intraperitoneal (i.p.) administration of 25 microg rtIFNgamma per chick 30 min prior to per os SE challenge resulted in a 35% reduction (P < 0.01) in SE organ invasion when compared with control (vehicle injected) chicks 24 hr post-SE challenge. However, i.p. administration of 2.5 microg rtIFNy per chick was not efficacious in reducing SE organ invasion. In Expt. 2 and Expt. 3, i.p. administration of 13.75 microg rtIFNgamma per chick 30 min prior to per os SE challenge resulted in significant reductions of 38.4% (P < 0.025) and 31.58% (P < 0.01), respectively, in SE organ invasion as compared with control chicks 24 hr post-SE challenge. Administration of 2.5 or 25 microg rtIFNgamma per chick i.p. had no effect on SE organ invasion in either Expt. 2 or Expt. 3 24 hr post-SE challenge. Additionally, i.p. administration of rtIFNgamma 30 min prior to SE challenge in Expt. 2 and Expt. 3 was not associated with protection against SE organ invasion when organ culture was performed 72 hr postchallenge. Further, the oral administration of 25 microg rtIFNgamma per chick was not efficacious in conferring protection against SE organ invasion at 24 or 72 hr postchallenge when this route of administration was evaluated in Expt. 2. Similarly, the subcutaneous administration of a potential repository injection of 13.75 or 25 microg rtIFNgamma per chick did not protect chicks against SE organ invasion when evaluated 72 hr postchallenge. These data indicate a potential acute immunostimulatory activity of rtIFNgamma in chickens experimentally challenged with SE. Further, these experiments, although preliminary, are suggestive of the potential involvement of IFNgamma in cell-mediated or innate mechanisms of protective immunity against salmonellosis in chickens.     

Efficacy of narasin in the prevention of necrotic enteritis in broiler chickens

The efficacy of narasin in the control of necrotic enteritis (NE) was investigated in a floor pen study of 2000 broiler chickens using a Clostridium perfringens feed inoculum challenge model. Treatments were 1) nonmedicated, nonchallenged; 2) nonmedicated, challenged; 3) narasin, nonchallenged; 4) narasin, challenged. Narasin was administered at 70 ppm in the feed from day 0 to trial termination on day 41. Challenge inoculum contained approximately 1 x 10(8) colony-forming units CP/ml and was administered from day 14 to day 16. In the unmedicated groups, challenged birds had significantly (P < 0.05) lower mean body weight and reduced feed efficiency at day 21 and significantly (P < 0.01) higher cumulative NE mortality at day 41 compared with unchallenged. Similarly, among unmedicated birds, those challenged had a significantly (P < 0.01) higher mean NE score on day 17 and significantly (P < 0.05) higher mean huddling scores on days 15-17 than unchallenged. Among challenged birds, those fed narasin had significantly (P < 0.05) higher mean body weight and improved feed efficiency at days 21 and 41 and significantly (P < 0.01) lower cumulative NE mortality at day 41 than unmedicated. Similarly, among challenged birds, those receiving narasin had a lower mean NE score on day 17 (P > 0.05) and significantly (P < 0.05) lower huddling scores on days 16 and 17 than unmedicated. Coccidiosis lesion scores were zero for birds euthanatized from all treatment groups on day 17, suggesting that the beneficial effects of narasin were not due to prevention of coccidiosis. This study thus provides evidence that narasin is effective in the prevention of necrotic enteritis in broiler chickens.     

Presence of inoculated Campylobacter and Salmonella in unabsorbed yolks of male breeders raised as broilers

Day-old male broiler breeder chicks were obtained from a commercial hatchery and raised as broilers. For Experiment 1, at 5 wk of age, the broilers were orally inoculated with a 10(6) cfu/ml of a characterized strain of Campylobacter jejuni and a cocktail (three naladixic acid-resistant strains) of Salmonella serovars. One week after inoculation, the birds were euthanatized and defeathered. The abdominal cavity was examined and any unabsorbed yolk material (and remaining yolk stalk) and ceca were aseptically removed for microbiological analyses. For each pooled sample (two birds per pool), an aerobic plate count (APC), an Enterobacteriaceae (ENT) count, and a test for the presence of Campylobacter and Salmonella was performed. For Experiment 2, at 5 wk of age, the broilers were orally inoculated with 10(5) cfu/ml of a characterized strain of Campylobacter jejuni. One week after inoculation, the birds (n = 20) were killed, defeathered, and the yolk stalk, attached yolk, or free-floating yolk and ceca were individually analyzed for presence of Campylobacter. For Experiment 1, the Salmonella-inoculated birds had 2/12 ceca and 0/12 unabsorbed yolk samples positive for Salmonella. The average yolk APC was log10 3.4 cfu/g and the average ENT was log10 1.9 cfu/g. For the Campylobacter-inoculated birds, 12/12 ceca and 9/12 unabsorbed yolk samples were positive for Campylobacter. The average yolk APC was log10 3.5 cfu/g and the average ENT was log10 3.1 cfu/g. For Experiment 2, the inoculated Campylobacter birds had 19/20 ceca, 5/20 free floating yolks, and 19/20 yolk stalks positive. In Experiment 1, the inoculated Campylobacter colonized the ceca in every instance and were present in 75% of the unabsorbed yolks. Alternatively, the inoculated Salmonella were not found in any of the unabsorbed yolks and only rarely in the ceca. In Experiment 2, the inoculated Campylobacter was found in very high numbers in the yolk and internal body samples. Determining to what extent these internal bodies and unabsorbed yolks play in bacterial colonization and contamination of the birds at processing has not been determined. The next step will be to determine the incidence of unabsorbed yolks and presence of Campylobacter and Salmonella in these bodies of commercial broilers at processing.     

Comparison of Salmonella enterica serovar enteritidis levels in crops of fed or fasted infected hens

Long-term feed withdrawal has been shown to increase ileocecal intestinal colonization and fecal shedding of Salmonella enterica serovar Enteritidis in challenged hens. Less information is available regarding effects of fasting on crop colonization. Two trials were conducted to compare effects of 14-day feed withdrawal vs. full feed on crop colonization in hens challenged with Salmonella Enteritidis. The levels of Salmonella Enteritidis in the crops of fasted hens were significantly higher than in nonfasted hens on days 3 and 10 and days 3, 9, and 16 postinfection (PI) in trials 1 and 2, respectively. Fecal shedding of Salmonella Enteritidis was significantly increased in the fasted hens on day 10 PI in trial 1. Analysis of crop IgA anti-Salmonella Enteritidis lipopolysaccharide levels in crop lavage samples of hens in trial 1 revealed a humoral response PI in both treatment groups with no significant differences, although peak response for fasted hens occurred 1 wk later. Histologic evaluation of hematoxylin and eosin-stained crop sections from trial 1 birds revealed mild to moderate heterophilic infiltration within the crop lamina propria (LP) or LP and epithelium of nonfasted infected hens at 24 and 96 hr PI. In comparison, heterophils in crops of fasted hens infected at this time point were sparse, indicating a possible diminished heterophil response in the fasted birds. Multifocal areas of tissue inflammation, as indicated by marked heterophil infiltration, with necrosis and sloughing of epithelium, were observed in crops from fasted hens at day 11 PI (14th day of feed withdrawal) but not in the fed groups. This severe heterophilic inflammation was observed in both challenged and nonchallenged fasted hens, suggesting that some factor other than Salmonella Enteritidis was responsible. These results indicate that feed withdrawal can have a dramatic effect on the integrity of the crop and its ultimate response to infection.     

Effect of dietary mannanoligosaccharide and sodium chlorate on the growth performance, acute-phase response, and bacterial shedding of weaned pigs challenged with Salmonella enterica serotype Typhimurium

A 28-d experiment evaluated the growth, acute-phase response, and bacterial shedding patterns in pigs (n = 96; initially 6.8 +/- 1.3 kg) fed mannanoligosaccharides (MANNAN) and sodium chlorate (CHLORATE) before and after oral challenge with Salmonella enterica serotype Typhimurium (ST). The negative control diet contained no antimicrobial (CON), and the positive control contained carbadox (CARB; 55 ppm). Test diets contained (as-fed basis) MANNAN (1,500 ppm) or CHLORATE (800 ppm). Pigs were fed diets for 14 d and then given ST orally. Pigs fed CARB had greater ADG over the entire study than pigs from other treatments (P < 0.05). During wk 1 to 2, before ST challenge, feed intake (as-fed basis) was lower for pigs fed MANNAN and CHLORATE than pigs fed CARB (P < 0.05). During the final 2 wk, pigs fed CARB had greater feed intake than pigs on other treatments (P < 0.05). Gain/feed was greater for pigs fed CARB in the 2 wk before ST (P < 0.05); however, in wk 3 to 4 after ST, gain/feed was reduced for CON pigs compared to pigs on other treatments (P < 0.05). Serum IGF-I was decreased at 2 and 4 d after ST (P < 0.001), and, overall, IGF-I was greater in pigs fed CARB than CON or CHLORATE (P < 0.05). Serum haptoglobin concentrations were greater (P < 0.001) for all treatments at d 6 compared with d 13 after ST. Overall, haptoglobin was greater for MANNAN than for CARB and CHLORATE (P < 0.05) and tended to be increased (P < 0.06) relative to CON. Interleukin-6 was not affected by treatment or day post-ST challenge. Fecal shedding of salmonellae organisms was less for CHLORATE (P < 0.05) than all other treatments at 7 d after ST. Shedding scores decreased from d 7 to 14 after ST (P < 0.05) for the CON, CARB, and MANNAN treatments. We conclude that feeding MANNAN and CHLORATE before acute enteric disease challenge may support improved gut function as evidenced by improved gain/feed, and that CHLORATE may decrease bacterial shedding. But neither MANNAN nor CHLORATE enhanced growth relative to the absence of dietary antimicrobials, nor was either treatment as effective as CARB following ST challenge.     

Efficacy of selected coccidiostats in sandhill cranes (Grus canadensis) following challenge.

The anticoccidial efficacy of amprolium, clazuril, and monensin were studied in sandhill cranes (Grus canadensis) infected with a mixture of Eimeria spp. oocysts. Five groups of four 1-day-old sandhill crane chicks were maintained on a crumbled ration containing no coccidiostat, amprolium at 2.2 ppm, clazuril at 1.1 ppm, clazuril at 5.5 ppm, or monensin at 99 ppm. After 2 wk on their respective feeding regimens, birds in each of the five groups were administered 25 x 10(3) pooled sporulated Eimeria spp. oocysts per os and observed for another 3 wk. A sixth group of four chicks served as nonmedicated, nonchallenged control during the study. Clinical signs and lesions consistent with disseminated visceral coccidiosis were observed in all challenged controls and birds fed amprolium and clazuril. Birds in these groups died 9-10 days after challenge. In contrast, only one monensin-medicated bird had clinical signs of disseminated visceral coccidiosis, and it died 13 days after challenge (DAC). This and an asymptomatic bird that were necropsied at study termination had less-severe gross and microscopic lesions of disseminated visceral coccidiosis. Two of three monensin-treated birds that survived challenge passed from 50 to 500 coccidial oocysts 11 to 18 DAC but were negative at study termination. Of the coccidiostats tested, monensin, at the dietary level of 99 ppm, was the only anticoccidial drug that provided protection against experimentally induced disseminated visceral coccidiosis in sandhill cranes.     

Cross-protection against Salmonella Typhimurium infection conferred by a live attenuated Salmonella Enteritidis vaccine

In this study, a genetically engineered live attenuated Salmonella Enteritidis (SE) vaccine was evaluated for its ability to protect against Salmonella Typhimurium (ST) infection in chickens. The birds were orally primed with the vaccine on the 1st day of life and given an oral booster at 5 wk of age. Control birds were orally inoculated with phosphate-buffered saline. Both groups of birds were orally challenged with a virulent ST strain at 9 wk of age. Compared with the control chickens, the vaccinated chickens had significantly higher levels of systemic IgG and mucosal IgA against specific ST antigens and a significantly greater lymphoproliferative response to ST antigens. The excretion of ST into the feces was significantly lower in the vaccinated group than in the control group on days 9 and 13 d after challenge. In addition, the vaccinated group had significantly fewer pronounced gross lesions in the liver and spleen and lower bacterial counts in the internal organs than the control group after challenge. These data indicate that genetically engineered live attenuated SE may induce humoral and cellular immune responses against ST antigens and may confer protection against virulent ST challenge.     

Effect of dietary beta1-4 mannobiose in the prevention of Salmonella enteritidis infection in broilers

1. This study investigated the effects of beta1-4 Mannobiose (MNB)-supplemented feeds on the kinetics of Salmonella enterica serovar Enteritidis (SE) in broilers and the ensuing histopathological changes. D-Mannose (MAN) was used for comparison. The diets supplemented with MNB or MAN were fed during the first two weeks after hatching to investigate any protection against SE infection in growing birds and any immunomodulatory functions in the gut. 2. MNB-supplementation reduced SE organ colonisation, caecal carriage and faecal shedding in a time-dependent manner. The high concentrations and persistency of the SE-specific IgA response in those birds given rations supplemented with MNB or MAN were associated with a decline in SE shedding and caecal carriage in the later stages of infection. MNB was more effective against SE infection than MAN. 3. Histological examination of the caecal wall and caecal tonsils at 23 d post-infection indicated a lesser degree of intestinal pathology. An increased number of intra-epithelial mononuclear cells (mature lymphocytes and macrophages) in the lining epithelium of birds fed on the diet supplemented with MNB was accompanied by an increased number of lamina propria cells. 4. The present study indicates that feeding a diet supplemented with MNB during the first two weeks after hatching reduced susceptibility to SE infection. Supplementing the diet with MNB or MAN increased IgA production and improved SE clearance by acting as immunomodulatory agents that prevented intestinal pathology. Feeding a MNB-supplemented diet to broilers could be used as an alternative to antibiotics, because it has no adverse effects on mortality or weight gain.     

Combination of competitive exclusion and immunization with an attenuated live Salmonella vaccine strain in chickens

To use the advantages of both the competitive exclusion (CE) technique and immunization with a live Salmonella vaccine, the combination of these methods was studied. Specific-pathogen-free chickens were pretreated by combined or single administration of a CE culture and a commercial live Salmonella typhimurium vaccine on days 1 and 2 of life and challenged with Salmonella typhimurium on day 3 to study the exclusion effect by both the CE preparation and the Salmonella vaccine. The exclusion effect by the CE culture combined with the immunologic effect by the live vaccine was studied after challenge of the birds on day 43 of age. The number of challenge organisms in ceca was used to evaluate the efficacy of the pretreatment. The protective exclusion effect of the CE culture was substantial in very young chicks and still detectable in 6-wk-old birds. The attenuated Salmonella typhimurium vaccine produced only an initially occurring exclusion effect. Because the exclusion effect of the CE culture was considerably stronger than the exclusion effect of the attenuated Salmonella typhimurium vaccine, the combination of both did not result in an additive protective effect. In order to exploit the exclusion potential between Salmonella strains and to attain an additive exclusion effect by a CE culture and a vaccine strain, live Salmonella vaccines are needed that are sufficiently attenuated without affecting genes essential for colonization exclusion of other Salmonella organisms. In 6-wk-old birds, the exclusion effect by the CE culture combined with the immunologic effect by the live Salmonella vaccine resulted in a degree of protection considerably beyond that generated by the exclusive use of the two methods. The administration of the live Salmonella vaccine strain prior to or simultaneously with the CE culture revealed the best protective effect because such combinations ensure an adequate persistence of the vaccine strain as prerequisite for the expression of an exclusion effect in very young chicks and the development of a strong immune response affording protection in older birds.     

Gamma/delta T cell response of chickens after oral administration of attenuated and non-attenuated Salmonella typhimurium strains

Poultry represents an important source of Salmonella infection in man. Despite intensive research on immunity, little is known about the involvement of T cell sub-populations in the immunological response of chickens against infection with non-host-adapted Salmonella (S.) serovars. In this study, the T cell composition of blood lymphocytes (CD4(+)CD8(+); CD4(+)CD8(-); CD4(-)CD8(+); CD8(+)TcR1(+); CD8(-)TcR1(+), CD8(+)TcR1(-)) after oral administration of the non-attenuated S. typhimurium wild-type strain 421 (infection) or the attenuated vaccine strain Salmonella vac((R)) T (immunization) to day-old chicks was investigated and compared with non-treated chickens by flow cytofluorometry. Additionally, the occurrence of T cell sub-populations (CD4(+); CD8(+); TcR1(+)(gammadelta); TcR2(+)(alphabeta(1))) in ceca, spleen and bursa of Fabricius of the birds was studied immunohistologically. Blood samples and tissues were examined between days 1 and 12 of age.Chicks inoculated with S. typhimurium 421 or Salmonella vac((R)) T showed significantly elevated percentages of CD8(+)TcR1(+) in blood on days 7, 8 and 9, or on day 8 in comparison to control animals. The CD4 to CD8 cell ratio was about 3:1 in infected animals on day 5 of age. In the organs of treated chicks the numbers of CD8(+)(gammadelta) and TcR1(+)(gammadelta) cells had markedly increased on days 4 and 5 in ceca, 8 and 9 in the bursa and 9 and 12 in the spleen. Moreover, infected or vaccinated birds revealed larger quantities of CD4(+) and TcR2(+) T cells in ceca on days 4 and 5. As shown by double staining, the TcR1(+) cells in the organs of infected animals additionally carried the CD8 antigen.In conclusion, immunization of day-old chicks with the attenuated Salmonella live vaccine strain resulted in the same changes in T cell composition as seen after infection with the non-attenuated Salmonella wild-type strain, but at a lower level. The remarkable increase of CD8(+)TcR1(+)(gammadelta) double positive cells in treated birds indicates an important role of this cell sub-population in the immunological defense of chickens against Salmonella exposure.     

Crop immune response post-Salmonella enteritidis challenge in eight commercial egg-layer strains and specific-pathogen-free White Leghorn chickens

The crop immune response against Salmonella Enteritidis (SE) challenge in eight commercial egg-layer strains (five white-egg layer and three brown-egg layer) and specific-pathogen-free (SPF) White Leghorn (WL) hens was investigated. Pre- and post-SE challenge mucosal immune responses within the crops were evaluated. Commercial layers and SPF WL hens were orally challenged with 10(8) CFU/ml SE PT13a and SE nalR PT13, respectively. Crop lavage samples were collected at weekly intervals from day 0 (pre-challenge) to day 25-27 postinfection (PI), and bacteriological examination was performed to monitor progression of SE infection. Crop lavage samples were analyzed for SE-lipopolysaccharide (LPS)-specific IgA using enzyme-linked immunosorbent assay (ELISA). H&E-stained slides of crop sections from day 34 PI and uninfected controls were assessed for lymphoid tissue via light microscopy. Lymphoid areas were graded based on morphology, size, and cellularity using a score 0 to 5 scale. The 0 to 5 (low to high) numerical values represented progressive increases in size and cellular density of lymphoid tissue. Bacterial culture results showed the highest percentage of SE-positive crop lavage samples from all hen groups at day 5-6 PI and day 11-12 PI. A progressive decline in percentage of SE-positive crop lavage samples did occur as time PI lengthened; however, at day 25-27 PI SE persisted in crop lavage samples from SPF WL hens and three commercial white-egg layer strains. A marked increase in SE-LPS-specific IgA was measured in crop lavage samples between day 0 and day 11-12 PI for all hen groups. Crop SE-LPS-specific IgA response remained elevated above day 0 baseline for the duration of the experiment. Well-defined score 3 to 5 lymphoid tissue aggregates were observed in crop tissue sections harvested at day 34 PI. Comparison of crop sections determined a 1.2-4.0 times increase in ratio of lymphoid tissue in day 34 PI SE-challenged hens vs. uninfected control hens.     

Laboratory evaluation of Newcastle disease vaccination programs for broiler chickens

Experiments were conducted to examine the efficacy of various commercial vaccination programs for the prevention of Newcastle disease (ND) in broilers. In all, chicks were from breeders vaccinated against ND via drinking water at 75-day intervals. Vaccination was by company personnel on company premises. In Expt. 1, the initial ND vaccination programs tested were vaccination at 1 day by coarse spray with the Spra-Vac machine or by tracheal instillation with the Beak-o-Vac machine, and vaccination at 7 days via drinking water. In Expts. 2-4, birds initially vaccinated via one of the three previously mentioned methods (Spra-Vac in Expt. 2, Beak-o-Vac in Expt. 3, and drinking water in Expt. 4) were revaccinated against ND by either drinking water or coarse spray with one of two commercial portable machines (ULVA Fan or Spray Master). Serologic and challenge data in Expt. 1 indicated that although broilers vaccinated by any of the three initial routes failed to produce measurable antibody to NDV, all methods resulted in protection against NDV challenge at 35 and 49 days. However, resistance to challenge with virulent ND was greatest in birds initially vaccinated by coarse spray with the Spra-Vac machine. Results in Expts. 2-4 indicated that NDV hemagglutination-inhibition titers were highest and resistance to challenge greatest in birds initially vaccinated at day 1 by coarse spray (Spra-Vac) and then revaccinated at 14 days by coarse spray. There were no differences, however, between the portable coarse spray machines in efficacy in reimmunizing broilers against NDV.     

Effect of feeding selected carbohydrates on the in vivo attachment of Salmonella typhimurium in chick ceca.

Two-day-old chicks were orally inoculated with 1 ml of Salmonella typhimurium (10(5) colony-forming units/ml) and divided into four groups. Three groups were fed 2.5% carbohydrates starting on day 1 (arabinose, galactose, and lactose), while the fourth group served as the control. Ceca were obtained from each group at 7, 14, and 21 days. At the end of 14 days, all three carbohydrates statistically reduced Salmonella recovery. However, lactose failed to reduce recovery between day 14 and day 21. Arabinose and galactose continued to show significant reductions of recovery through 21 days. No statistical difference was found between Salmonella recovery from whole ceca (with cecal material) and inverted ceca (washed free of cecal material).     

Effect of dietary inclusion of spray‐dried porcine plasma on performance,some physiological and immunological response of broiler chickens challenged with Salmonella sofia

This study was conducted to investigate the effect of spray‐dried porcine plasma (SDPP) in broiler chickens under Salmonella sofia disease challenge. The experiment comprised five starter diets: positive control (no supplement), diet supplemented with in‐feed antibiotics (IFA; salinomycin 0.05% + zinc bacitracin 0.033%) and diets supplemented with SDPP at 10 or 20 g/kg diet. All four of these groups were challenged with S. sofia, while a fifth group was unchallenged and used as the negative control. The experimental diets were fed to 14 days; then, the birds were switched to commercial‐type grower and finisher diets. Oral inoculation of the challenged groups with S. sofia occurred on day 8, 10 and 12. Body weight was significantly higher in the birds fed diets containing IFA and SDPP than in the challenged control group, but it was only significant in starter and grower phases. In general, there was an improvement in the weights of the immune‐related organs, but it was only significant for the weight of the bursa of SDPP‐fed birds at 13 days. At day 13, blood potassium content was lower and the concentrations of IgG and IgM tended to be lower in the birds fed on low‐SDPP starter diets than those of the other groups. There were significant differences in the concentration of lactic acid in the ileum and acetic acid, formic acid, butyric acid and propionic acid in the caeca. Inclusion of SDPP to the starter diets of broiler chicks had positive effects on broiler performance, immunity and gut health during exposure to highly pathogenic conditions.     

Evaluation of a French ELISA for the detection of Salmonella Enteritidis and Salmonella Typhimurium in flocks of laying and breeding hens

In France, the regular and compulsory detection of Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) in flocks of breeding and laying hens is based on bacteriological examination of environmental swabs and faeces samples. The aim of this study was to compare this bacteriological examination with a serological method (ELISA) developed in our laboratory. This ELISA was first evaluated by use of artificially infected hens. During these experimental infection studies, several groups of hens were inoculated with SE, ST, different vaccines and different Salmonella serovars to calculate the experimental parameters of our ELISA. Then, in a field study, 43 flocks were followed monthly using two bacteriological samples (environmental swab and pool of faeces) and 20 serological samples (sera or yolks). Twenty-seven flocks without SE or ST gave a negative serological response throughout their surveillance. Among the 10 various serovars different from SE and ST isolated in this study, S. Heidelberg, S. Agona and S. Hadar gave seropositive results in seven flocks. Consequently, this ELISA was not specific of SE and ST as it detected serovars sharing or not common antigens with SE and ST. Seropositive results were also obtained each month for two flocks where no Salmonella could be isolated. Finally, in seven flocks found infected with SE or ST, the positive ELISA results appeared later than the bacteriological detection. Therefore, for the detection of chicken flocks recently infected with SE or ST, bacteriological examination currently used in France seems to be more appropriate than this ELISA.