Salmonellosis in Turkeys: Evaluation of Bacteriological and Serological Evidence of Infection

Bacteriological and serological studies were made of Salmonella infections in turkeys under three conditions; in commercial hatchery supply flocks and their embryo and poult progeny, in survivors of a natural infection over a ten-month period, and in experimentally infected adult turkeys.

The investigations revealed the following: 1. Carrier birds yielded Salmonella from cloacal swabs only intermittently which emphasized that failure to so isolate the organism did not necessarily indicate absence of infection in flocks. On the other hand, recovery of Salmonella from swabs or from dead-in-shell embryos foretold the likelihood of clinical disease in the progeny.

2. Bacteriological examination of ten per cent of the embryos from infected individual hatches did not regularly yield Salmonella but culture of a series of hatches revealed the disease status of the progeny.

3. The percentage of infected embryos in one hatchery which frequently disseminated S. typhimurium was estimated to range from two to twenty per cent in different hatches.

4. Environmental sanitation in the flock and hatchery appeared to be a major influence in the frequency of clinical outbreaks in the poults.

5. Two hatchery supply flocks were found to be infected with at least two serotypes; a third serotype appeared in the progeny of one in addition to the two in the parent flock.

6. Hatchery supply flocks with no history of clinical infection but having a latent infection transmitted the disease as readily as those which had survived clinical outbreaks.

7. The overall performance of the bacterial agglutination and hemagglutination tests demonstrated an equal but unsatisfactory degree of efficacy insofar as detection of infected birds was concerned. Both tests failed to identify half of the infected birds and also one infected flock.

8. A high incidence of non-specific reactions was observed in the hemagglutination tests of sera from non-infected birds. The number was greatly reduced by selecting strains whose extracts were more suitable for modifying erythrocytes. The possibility of the practical application of this test as a screening procedure would appear to depend on overcoming the problem arising from frequent non-specific reactions.

     

Detection of Campylobacter jejuni strains in the water lines of a commercial broiler house and their relationship to the strains that colonized the chickens

Campylobacter jejuni is frequently present in the intestinal tract of commercial broiler chickens, and their drinking water has been proposed to be an initial source of bacteria for newly hatched chicks. We studied three sequential commercial broiler flocks raised in a house from which we had cultured C. jejuni from the nipple waters prior to placement of the first flock. Campylobacter cells were detected by immunofluorescence in the biofilm of the drinking nipples during the weeks when the flock was colonized with C. jejuni but not during weeks when the birds were negative. Campylobacter jejuni was isolated from the drinking water during the growth of the first flock and was present in the birds from all three flocks. Randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing with primer OPA11 indicated that seven distinct strains were present within the broiler house. One strain found in drinking water was similar to a strain found in birds in the second flock; however, RAPD-PCR with primer HLW85 showed that the strains were not identical. These results suggest that although the watering system is a potential source of C. jejuni in broiler flocks, the waterborne strain in this study was not detected in the birds.     

Alterations in levels of various host antioxidant factors in turkey knockdown syndrome

Five components (selenium, glutathione peroxidase, copper, superoxide dismutase, and vitamin E) of the antioxidant system of turkey poults were examined to determine if they play any role in the knockdown (KD) syndrome. All flocks were provided with feed formulated to contain monensin at 54-60 g/ton. Flock data were analyzed as a case-control study with three treatment groups (KD-affected and unaffected turkey poults from a KD flock and poults from an unaffected flock [control]). Affected turkey poults had lower (P < 0.001) serum vitamin E levels compared with unaffected poults from KD flocks or poults from unaffected flocks. No significant differences were observed for the other parameters evaluated but there was a trend towards lower copper and superoxide dismutase values in affected birds. It appears that serum vitamin E concentrations in turkey poults may play a significant role in susceptibility to or protection against KD syndrome. Other components of the antioxidant system may also be involved, and complex interactions among several body systems may be critical.     

Serological investigations of infectious bursal disease virus and reticuloendotheliosis virus infections in New Zealand chickens

Extract

Sir:- Between 1975 and 1981, sera were collected from 89 New Zealand chicken farms representing a variety of laying and meat birds of different ages from both primary breeders and commercial flocks. In some instances samples were collected from one or more flocks on a farm. In most cases between ten and 20 birds were sampled in a flock and the sera pooled to give a composite sample for testing.     

The effects of early and late maturity on the protein requirements of pullets

Growing pullets were exposed to two light patterns which caused a difference in sexual maturity of 5 weeks. From 28 to 38 weeks of age they were fed diets containing crude protein levels of 7.0, 8.5, 10.0, 11.5, 13.0 and 14.5 per cent. Yellow maize provided 45 per cent and soyabean meal 55 per cent of the protein in all six diets.

Rate of lay, egg weight and body weight were greater in the late maturing flock than in the early flock at the start of the assay and throughout the assay period. The late maturing pullets required more protein to reach and maintain their maximum potential than the early birds, presumably because their potential output was greater. The late birds showed a diminishing but continued response to protein up to the highest level fed (14.5 per cent corresponding to 23 g. protein per bird day). The early maturing pullets showed no response to dietary protein levels beyond 11.5 per cent and their estimated protein requirement was about 16 g. per bird day.

When limiting amounts of protein were fed (7.0–10.0 per cent of the diet) the two flocks achieved similar levels of egg output. In both flocks and throughout the assay, body weight and egg weight fell at the two lowest levels and increased at the three highest levels of protein.

It is concluded that the protein requirements of early maturing and late maturing flocks of pullets may differ, but only in a way which corresponds directly with their different potential outputs. In the late flock, which had the higher protein requirement, excellent production was obtained with a diet containing 14½ per cent crude protein all of which came from vegetable sources.     

Infectious Bursal Disease in New Brunswick

A flock of four week old chickens experienced a disease of sudden onset in which the only symptoms were those of depression shortly before death, and in which the predominant histological lesion was necrosis of lymphocytes in the bursa of Fabricius.

A virus, designated strain Sk-1, was isolated from pooled bursal tissue of affected birds and was serologically identified as a strain of the infectious bursal agent. This virus was chloroform resistant, did not hemagglutinate guinea pig or chicken erythrocytes and did not produce a cytopathic effect in chick embryo tissue cultures. Equivocal results were obtained in filtration studies but the agent was less than 100nm in diameter.

Four week old chicks inoculated with strain Sk-1 developed microscopic lesions in the bursa of Fabricius which were similar to those seen in the original field specimens. Inoculated chick embryos exhibited characteristic macroscopic lesions and necrosis of vascular tissue was a common histological change.

A limited serological survey of local poultry flocks indicated that infection by this agent had occurred in four of the ten flocks examined.

     

Studies on Effect of Lighting on “Sudden Death Syndrome” in Broiler Chickens

Broiler chicken flocks were studied to determine the mortality from sudden death syndrome occurring in the flocks. The difference in the incidence of the syndrome in pullets and cockerels, and the age at which the most birds are affected were also studied. The weight of sudden death syndrome birds was compared with the flock average and the effect of continuous lighting as opposed to intermittent lighting was examined.

The results suggest that; continuous lighting produces more sudden death syndrome deaths than intermittent lighting; that the incidence of sudden death syndrome is higher in cockerels than pullets; that the highest death rate occurred during the third and fourth weeks of life, and that sudden death syndrome birds on the average were heavier than the flock average.

     

Isolation of Salmonella Organisms from Commercial Layer Houses Where the Flocks Were Molted with a Wheat Bran Diet

To develop an alternative method to feed withdrawal for molting layers, 2 flocks consisting of approximately 26,000 commercial laying hens each at 478 (68 wk, flock 1) and 466 (67 wk, flock 2) d of age were reared in an environmentally controlled windowless house and were fed wheat bran (WB) diet. Flock 1 hens were fed WB for 25 d, and flock 2 hens were fed WB for 21 d and then fed a mixture of WB and layer feed (1:1, wt:wt) for the last 4 d of the treatment. After that, the birds in both flocks were fed a normal layer feed. The photoperiod was reduced from 16 to 9 h in both flocks. Most of the birds in both flocks ceased egg production by 10 to 15 d of feeding the WB diets. Egg production in flock 1 gradually increased to 11.4% by 31 to 40 d and 71.4% by 41 to 50 d of the treatment, whereas the egg production in flock 2 hens lagged behind by almost 10 d. The mean egg production from 61 to 140 d exceeded 86% in both flocks. The houses in the farm were naturally contaminated with several serovars of Salmonella, not Enteriditis or Typhimurium. In both flocks with the WB treatment, no marked increase in Salmonella isolation from environmental samples was observed postmolt relative to premolt levels. The study demonstrated that feeding hens WB could be successfully used as an alternative to feed withdrawal to force-rest aging hens while not exacerbating a Salmonella problem in a commercial egg-production setting.     

Experimental transmission of turkey viral hepatitis to day-old poults and identification of associated viral particles resembling picornaviruses.

Turkey viral hepatitis (TVH) was experimentally reproduced in two experiments in 1-day-old poults. In the first experiment, an infectious inoculum was prepared from filtered yolk materials harvested from dead embryonating chicken eggs (ECE) previously inoculated with suspensions of liver and pancreas tissues collected from TVH-affected birds in commercial turkey flocks. One-day-old poults given a yolk-sac inoculation or oral gavage with this preparation developed lesions in the liver and pancreas characteristic of TVH at 20 days postinoculation (PI) in 60% and 14% of the experimentally infected birds, respectively. With the identical inoculum, embryo mortality occurred at 8 and 10 days PI in embryonating turkey eggs (ETE) inoculated into the yolk sac. In the second experiment, an infectious inoculum was prepared from filtered yolk materials from dead ETE harvested in the first experiment. One-day-old poults given a yolk-sac inoculation with this filtered yolk material developed lesions in the liver and pancreas within 5 days PI. At 20 days PI, 67% of the experimentally infected birds had similar lesions. With the inoculum given to these poults, embryo mortality occurred at 6, 8, and 10 days PI in ETE inoculated into the yolk sac. Virus particles 26-28 nm in diameter with icosahedral morphology typical of picornaviruses were identified by EM in the yolk sacs of ETE that died in both experiments, and inoculated ETE that died following passage of filtered suspensions of pancreatic tissues collected from affected birds in the first experiment.     

Sequential spread of Campylobacter infection in a multipen broiler house

Generally, colonization with Campylobacter jejuni is first detected in broilers 2-3 wk after hatching. Once introduced into a flock, this infection spreads very rapidly. The sources and routes of transmission of C. jejuni in broilers remain debatable. In this study, the spread of infection was monitored in a commercial multipen broiler house in which birds were contained in discrete groups and sampled sequentially. Colonization was monitored in two broiler flocks up to slaughter. Serotyping and fla typing methods were applied to differentiate all the C. jejuni strains isolated. In flock 1, colonization was first detected at 32 days of age in birds located at the rear of the house. By 40 days, nearly all the birds were infected with the same strain (fla type 1.9). However, at 46 days of age, a second strain (fla type 3.7) was detected in some of the birds. These birds were also located toward the rear of the house. In flock 2, infection was detected at 5 wk of age. This infection was once again first detected in birds located at the rear of the house. In this flock, only a single fla type (1.1) was isolated throughout. A survey of the broiler house relative to the location of first point of infection indicated the use of an entrance door unprotected by boot dips. However, securing this door during the second flock study did not prevent infection.     

Vitamin A deficiency in turkey poults.

Vitamin A deficiency was diagnosed in a commercial flock of 13,000 4-6-week-old turkey poults in the summer of 2004. The birds were initially submitted for examination because of a 3% increase in the reported daily mortality of the flock. Clinically, affected birds had stunted growth and ruffled feathers, showed signs of incoordination, and were depressed. At necropsy, pale white pseudomembranous to mucoid material was observed on the mucosal surface of the tongue, oral cavity, portions of the esophagus, and the crop of some birds. Histologically, there was squamous metaplasia of the mucosal epithelium of the oral mucosa, esophagus, sinuses, nasal glands, bronchi, proventriculus, and the bursa of Fabricius. Vitamin A was not detected in the feed sample at a detection limit of 0.5 mg/kg. Serum vitamin A concentrations in 7 birds were very low and ranged from 0.05 to 0.1 mg/L. Vitamin A concentrations in livers were extremely low (0.1 mg/kg wet weight, 1/7 poults) or undetectable (< 0.1 mg/kg wet weight, 6/7 poults). A diagnosis of vitamin A deficiency was made based on gross and microscopic lesions and vitamin A concentrations in serum, liver, and feed. To the authors' knowledge, this is the first documented case of vitamin A deficiency in poults submitted from a commercial meat turkey producer comparatively depicting the gross and microscopic lesions with those found in other species of birds and mammals.     

Seroprevalence of Histomonas meleagridis in pullets and laying hens determined by ELISA

Serum samples from 1120 layers from 56 flocks and 400 pullets from 20 flocks were tested by an indirect sandwich ELISA to investigate the prevalence of antibodies to Histomonas meleagridis in chickens kept in alternative husbandry systems. The overall prevalence of antibodies to H meleagridis in layers was 37.3 per cent, and positive birds were identified in 50 flocks. This was significantly higher than in pullets, where only 8.3 per cent of the birds tested positive. Optical density (OD) values obtained from pullet sera were much lower than the OD values from layers; however, positive birds were detected in half of the pullet flocks. In particular, all birds from an organic pullet flock were found to be positive, with high OD values. Overall, the highest prevalence of positive sera was obtained from birds kept in free-range flocks. Attempts to reisolate live histomonads from birds in 18 layer flocks were unsuccessful.     

The effect of increased mineral levels in the feed on leg weakness and sudden death syndrome in broiler chickens

In two experiments with 98,000 and 40,000 broilers on commercial broiler farms, half of each flock were fed starter, grower and finisher diets with 0.2% added calcium, 0.2% added phosphorus and 0.2% added magnesium. This ration had no significant effect on the incidence of sudden death syndrome but the incidence of leg weakness in broilers on the increased mineral ration was higher in both flocks.

Approximately 28% of the mortality in both flocks was from sudden death syndrome, more than from any other condition. Approximately 20% of the mortality was associated with leg weakness.

     

Persistence of immunity in commercial egg-laying hens following vaccination with a killed H6N2 avian influenza vaccine

The California poultry industry experienced an outbreak of H6N2 avian influenza beginning in February 2000. The initial infections were detected in three commercial egg-laying flocks and a single noncommercial backyard flock but later spread to new premises. The vaccination of pullet flocks with a commercially prepared, killed autogenous vaccine prior to their placements on farms with infected or previously infected flocks was used as a part of the eradication programs for some multiage, commercial egg production farms. The purpose of this study was to follow three vaccinated flocks on two commercial farms to track the immune responses to vaccination. The antibody-mediated responses of the three flocks followed in this study were markedly different. One flock achieved 100% seroconversion at 12.5 wk of age, but by 32 wk of age, all of the hens were seronegative by agar gel immunodiffusion (AGID). In contrast, at 32 wk of age, flocks from the other farm (flocks 2A and 2B) were 95% and 72% seropositive by AGID, respectively. Of the differences that were identified between the vaccination protocols on the two farms, the distinction that could explain the level of disparity between responses is the delivery of the second dose of vaccine with a bacterin on the first farm, which may have interfered with the persistence of immunity in this flock. Hens from flocks 2A and 2B were experimentally challenged at 25 wk of age with H6N2 avian influenza virus. Hens from flock 2A did not transmit virus to naive contact-exposed hens, but hens from flock 2B did. At 34 wk of age, hens from flock 2A were again challenged and naive contact-exposed hens were infected in this second trial. These challenge experiments served to demonstrate that despite detectable antibody responses in flocks 2A and 2B, the birds were protected from infection for less than 21 wk after the second vaccination.     

Development and application of an ELISA for detecting antibodies to Salmonella enteritidis in chicken flocks.

Enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of IgG antibody to Salmonella enteritidis in poultry flocks. A lipopolysaccharide (LPS) and heat-extracted (HE) antigen for use in the ELISA were evaluated together with the rapid slide test (RST), microagglutination test (MT) and the microantiglobulin (MAG) test. In experimentally infected specific pathogen free chickens, good correlation was seen between all tests although, generally, the MT and MAG detected antibody earlier and titres peaked earlier than the ELISAs. The LPS antigen detected antibody earlier than the HE antigen but the latter gave higher titres in the later stages of infection. Cross reactions were seen between S enteritidis and S typhimurium in the ELISAs although homologous reactions were always much higher. Antisera to S montevideo or S senftenberg gave weak positive reactions in both S enteritidis ELISAs. Serological and bacteriological examinations of representative samples from two commercial chicken flocks were carried out. In flock A the HE-ELISA and MAG test detected antibody in nearly all birds. The LPS-ELISA detected antibody in over 60 per cent of birds, while the MT and RST detected few seropositive birds. The whole blood test using the stained S pullorum antigen on the farm detected antibody in just under 25 per cent of the birds. S enteritidis was isolated from the organs of 25 per cent of the birds. All birds in flock B were seronegative by all tests; no salmonellae were isolated from the organs of these birds.     

Characteristics of fowl cholera outbreaks in turkeys in Georgia in 1986

Information was gathered from 64 cases of fowl cholera (FC) in turkey flocks through diagnostic case records, flock records, and telephone and mail surveys. Forty-five cases came from flocks of commercial turkeys, of which 15 were presented twice, and four came from mature breeder flocks. The prevalence of FC was 18.0% of commercial flocks and 14.7% of breeder flocks at risk. The average age at first diagnosis of FC was 90 days in commercial turkey flocks and 32 weeks 5 days in breeder flocks. Acute mortality was the most common presenting complaint, with a 0.37% average mortality in commercial flocks on the day of first presentation, 0.80% in commercial flocks presented a second time, and 0.43% in breeder flocks. Pasteurella multocida was cultured from 69.8% of the 361 tissue samples submitted from these cases. Novobiocin, penicillin, and chlortetracycline (CTC) had the greatest in vitro activity against isolates. Serotype 3-cross-4 was found in all 18 commercial flocks from which isolates were typed. All breeder flocks and 88.6% of commercial flocks were vaccinated before disease onset. Flocks were treated for an average of 14.3 days, most commonly with high levels of sulfadimethoxine and/or CTC. Body weights of affected birds were comparable to those of birds in unaffected flocks, but mortality and feed efficiency were worse.     

Cases of swollen head syndrome in broiler chickens in Greece

From 50 commercial broiler flocks included in a study concerning respiratory disease, signs of swollen head syndrome (SHS) were shown in eight. Postmortem examination was performed in eight birds showing signs of SHS from each flock. The trachea and head from each bird were collected for laboratory investigation. An enzyme-linked immunosorbent assay (ELISA) was used for the detection of viral and avian mycoplasma antigens in the trachea, and bacteriologic examinations were performed from the infraorbital sinuses of the infected birds. According to the ELISA results, the most frequently detected antigen in the trachea was Mycoplasma synoviae (six flocks, 75%), followed by infectious bronchitis virus (IBV) (five flocks, 62.5%), avian adenovirus (four flocks, 50%), avian reovirus (three flocks, 37.5%), Mycoplasma gallisepticum (one flock, 12.5%), and Newcastle disease virus (NDV) (one flock, 12.5%). Turkey rhinotracheitis (TRT), infectious laryngotracheitis, and avian influenza viral antigens were not detected. Experimental assays for characterization of NDV and IBV isolates showed that they were strains of low virulence (evidently vaccine strains). Bacteriologic examinations from the infraorbital sinuses of the affected birds resulted in the isolation of Escherichia coli (seven cases, 87.5%) and Staphylococcus spp. (one case, 12.5%). It is evident that TRT virus did not play a causal role in SHS in commercial broiler flocks in Greece, but in this condition, other viruses (IBV, NDV), mycoplasmas, or bacteria may be involved, and environmental conditions seem to be essential to the occurrence and severity of the disease.     

A survey of skeletal disorders in five turkey flocks in Saskatchewan.

Losses due to skeletal disorders varied from 1.8 to 6.1 percent in five flocks, varying in size from 2040 to 7033 birds, of broad breasted white tom turkeys. Very little evidence of significant air-sacculitis due to Mycoplasma meleagridis was found in hatching poults or during the growing period. Major disorders causing the losses included rickets, rotated tibia, arthritis/osteomyelitis and long bone distortion. Rickets and arthritis/osteomyelitis due to infection with Staphylococcus aureus were significant problems only in one flock each, while rotated tibia caused significant losses in three flocks. Long bone distortion was the major cause of economic loss and occurred in all flocks. Possible causes of long bone distortion are briefly discussed.     

Measurements of footpad dermatitis in broiler chickens at processing plants

A four-point photographic scale was developed to score the severity of lesions of footpad dermatitis (FPD) in broiler chickens. There was a linear relationship between the square root of the relative area of the lesions and their scores, and good agreement between different assessors using the scale. The scale was used to assess samples of 100 birds from each of 190 flocks slaughtered at two uk processing plants in 2002 and 2003; 12 of the flocks (6.3 per cent) had no lesions, but the others had lesions of different prevalences and severity. The maximum proportion of affected birds in a flock was 92 per cent. In the 178 affected flocks, 84.0 per cent of the birds had no lesions and 16.0 per cent had some evidence of FPD. The overall unweighted prevalence of birds with FPD in all 190 flocks sampled was 18.1 per cent; 10.2 per cent had only small lesions, on average equivalent in area to 2.1 per cent of the total area of the foot, 6.2 per cent had lesions on average equivalent to 6.6 per cent of the area of the foot, and 1.7 per cent had lesions on average equivalent to 21.5 per cent of the area of the foot. There were differences between the two plants in the overall prevalence and severity of FPD, but this may have been due to the fact that the plants were sampled at different times of the year.     

Use of an enzyme-linked immunosorbent assay to measure antibody levels in turkey breeder hens, eggs, and progeny following natural infection or immunization with a commercial Bordetella avium bacterin

Twelve large white turkey hens were immunized with a commercially available Bordetella avium bacterin. Hens and eggs were tested using an enzyme-linked immunosorbent assay (ELISA) to determine the response to the bacterin. Three hundred poults were then obtained from two commercial flocks, the hens of one flock having been immunized with the same bacterin used on the group of 12 turkeys. Titers of the poults were monitored for 7 weeks, and poults were challenged by exposure to infected poults at 1, 7, 14, and 21 days post-hatch. Hens produced an antibody response following immunization, with a parallel antibody response being detected in eggs. Maternal antibodies were present in poults from immunized hens. Poult titers declined to near the level of poults from unimmunized hens by 14 days of age. Poults from immunized hens challenged at 1 and 7 days were resistant to development of clinical disease and gross lesions, whereas all poults from unimmunized hens exhibited clinical signs and gross lesions. After 14 days, the resistance of both groups to development of clinical disease, became near equal, neither group being affected as severely as the unimmunized hens challenged at days 1 and 7. Six commercial turkey breeding flocks and their progeny that had not been vaccinated for B. avium and had no history of B. avium infection were evaluated with the B. avium ELISA. There were variations between the flocks, with poult titers reflecting those found in the hens.