Reovirus tenosynovitis in a flock of layer breeders.

The present paper describes a reovirus infection with clinical course in a flock of layer breeders. Lameness and tenosynovitis of flexor tendons were observed in approximately 15% of the cockerels and 3% of the hens from 17 weeks of age onwards. Affected birds did not die; on the contrary, most of them recovered clinically within a period of 8 weeks. Two other breeds of layer parents that were housed in close contact with the affected flock did not develop clinical signs, although serology indicated that infection with reovirus had taken place. These field observations constitute the first report of clinical reovirus tenosynovitis in layer parents and indicate different susceptibilities of layer parent breeds in developing clinical signs following reovirus infection.

Evaluation of the enzyme-linked immunosorbent assay for the detection of antibodies against infectious bursal disease virus (IBDV) and the estimation of the optimal age for IBDV vaccination in broilers.

In order to evaluate the usefulness of the enzyme-linked immunosorbent assay (ELISA) in determining the optimal age for infectious bursal disease virus (IBDV) vaccination in broilers under field circumstances, the percentage of birds susceptible to vaccine at vaccination age was estimated in 12 flocks on the basis of ELISA titres in newborn chicks. The effective percentage of susceptible birds at the day of vaccination was checked by comparing virus neutralization (VN) titres measured in sera taken immediately before vaccine administration with the breakthrough titre of the vaccine used. In nine of the 12 flocks, the ELISA had largely overestimated the percentage of susceptible birds. Subsequently, to examine the performance of the ELISA kit, 815 chicken sera were comparatively examined in ELISA and VN tests. Results demonstrated a linear relationship between mean ELISA titres and VN titres. It was concluded that the ELISA could be a useful tool in IBDV vaccination timing, but that interpretation criteria should be adjusted to avoid vaccination at too early an age.

Colibacillosis in caged layer hens: characteristics of the disease and the aetiological agent.

In Europe, outbreaks of acute mortality in layer flocks due to colisepticaemia have frequently been observed since the mid-1990s. The aims of this study were to describe the disease, to identify the serotypes of the avian pathogenic Escherichia coli (APEC) present in these outbreaks, and to detect the presence of F11 fimbriae and flagella in the isolates. For this purpose, 20 flocks with APEC-associated increased mortality and 20 control flocks matched for age were examined. Weekly mortality rates in the colibacillosis-affected flocks reached 1.71%, versus 0.30% in the control flocks. The maximum cumulative mortality over an entire colibacillosis outbreak reached 9.19%. The disease was often flock and hen house associated, with recurrent outbreaks within one round and in successive rounds in the same house. Disease was usually acute without clinical symptoms. Peritonitis with yolk material deposited in the peritoneal cavity and polyserositis were the main lesions at necropsy. O78 strains were isolated in 15 of the 20 colibacillosis flocks, and in only one of the control flocks. The majority of strains from the control flocks could not be serotyped by the 28 O-antisera used. In general, F11 fimbriae and flagella were present in the majority of the strains. F11 fimbriae were significantly more often found in O78 isolates than in the other serotypes, and are thus more often present in isolates from colibacillosis flocks. Strains positive for F11, and for F11 and flagella, were more frequently present in heart and liver of the colibacillosis-affected flocks.

Significance of interactions between Escherichia coli and respiratory pathogens in layer hen flocks suffering from colibacillosis-associated mortality.

This study aimed to examine the significance of interactions between Escherichia coli and various respiratory pathogens during outbreaks of colibacillosis-associated mortality in layer hen flocks under field conditions. For this purpose, a case-control study involving 20 control flocks with baseline mortality and 20 flocks with increased mortality due to E. coli septicaemia and polyserositis, was conducted. In each colibacillosis flock, blood samples were taken from 20 hens at the onset of clinical disease and three times thereafter at 2-week intervals. Control flocks of comparable ages were sampled in the same way. Pooled sera, taken at the first and last sampling, were examined for antibody titres against infectious bronchitis virus (IBV) and Newcastle disease virus (NDV), and the individual sera from all four samplings were examined for the presence and/or titres of antibodies against avian pneumovirus (APV), Mycoplasma gallisepticum, Mycoplasma synoviae and Ornithobacterium rhinotracheale. Titre increases were seen for IBV D274 (one control flock) and O. rhinotracheale (one control and one colibacillosis flock). An increase in per cent reactors was seen for APV (one control flock), and for M. synoviae (one control and two colibacillosis flocks). The study failed to detect any consistent interactions between E. coli and the aforementioned pathogens. These results indicate that, at least as observed in this study, outbreaks of increased mortality resulting from colibacillosis are not necessarily associated with IBV, NDV, APV, M. gallisepticum, M. synoviae or O. rhinotracheale infections.

Risk factors associated with colibacillosis outbreaks in caged layer flocks.

Colibacillosis appears to be of increasing significance in layer flocks, but there have been no studies of the risk factors associated with outbreaks. This study aimed to investigate the possible associations between risk factors of non-infectious nature and outbreaks of mortality due to colibacillosis in flocks of caged layer hens. Information on management, biosecurity measures and housing conditions was collected in 20 flocks suffering from the disease and in 20 clinically healthy control flocks. The data were processed using multiple logistic regression. The statistical analysis demonstrated that an increase in the distance to the nearest poultry farm by 1 km was associated with a six-fold decreased risk of an outbreak of colibacillosis (odds ratio=0.16). Furthermore, a 1 l increase in cage volume per hen was associated with a 33% decrease in the risk of an outbreak (odds ratio=0.75). It was concluded that the distance between poultry farms and the hen density in the cages are important risk factors for outbreaks of colibacillosis in flocks of layer hens.

An outbreak of histomoniasis in free-range layer hens.

Histomonas meleagridis was held primarily responsible for an outbreak of 6% increased mortality and 11% decreased egg production between weeks 57 and 72 in a flock of free-range layer hens, concurrently infected with Brachyspira-like bacteria. This case can be considered an example of ancient diseases re-emerging in alternative housing systems that are promoted because of animal welfare considerations, but that at the same time allow rapid spread of pathogens in birds for which only few curative drugs are registered. Therefore, new housing methods should be introduced gradually to gain experience with them.

Failure of a low virulence Streptococcus gallolyticus serotype 1 strain to immunize pigeons against streptococcosis.

Ten pigeons were inoculated intravenously with the low virulence Streptococcus gallolyticus strain PDH 827, which belongs to serotype 1, supernatant phenotype A(-)T2. The birds did not develop clinical disease but shed S. gallolyticus in their faeces, and antibodies against the bacterium were detected in post-inoculation plasma samples of all birds. Seven weeks later, these pigeons, as well as 14 control pigeons, were challenged intravenously with the highly virulent S. gallolyticus strain STR 357, which also belongs to serotype 1 but to the supernatant phenotype A(+)T1. Post-inoculation morbidity in the immunized group amounted to 90%, demonstrating that no protective immunity had been built up after the first infection. These findings indicate that serotype-specific antigens are not, or at least not solely, involved in induction of protection against S. gallolyticus septicaemia in pigeons.

Interactions of Salmonella enterica serovar Muenchen with macrophages of the turtle Trachemys scripta scripta.

Interactions of Salmonella with macrophages have been studied in birds and, most extensively, in mammals. In these homeothermic animals, interactions between Salmonella and macrophages are characterized by the following processes. After macropinocytosis, spacious phagosomes are formed within the macrophage. Partial inhibition of phagosome-lysosome fusion and resistance to the formation of reactive oxygen species and reactive nitrogen intermediates enable the bacterium to survive and even multiply within the host macrophage. Eventually, Salmonella will induce apoptosis of the macrophage. In this study, interactions of peritoneal macrophages of the turtle Trachemys scripta scripta with Salmonella enterica serovar Muenchen were examined in vitro. Turtle macrophages were able to phagocytise Salmonella efficiently at both 30 and 37 degrees C. Exposure of macrophages to Salmonella induced the production of reactive oxygen species, which could be partially suppressed by adding the NADPH oxidase inhibitor diphenylene iodonium. Initially, most of the intracellular bacteria were killed. However, Salmonella proved to be able to persist and multiply inside turtle macrophages at both 30 and 37 degrees C for at least 48 h, despite the production of reactive nitrogen intermediates by inducible NO synthase. Salmonella infection of turtle macrophages killed the phagocytes at both 30 and 37 degrees C. These findings demonstrate that no obvious qualitative differences exist between macrophage-Salmonella interactions from homeothermic animals and from turtles. This indicates that other factors are responsible for the different course of Salmonella infections in homeothermic and poikilothermic hosts.

The effect of Ornithobacterium rhinotracheale vaccination of broiler breeder chickens on the performance of their progeny.

The effect of Ornithobacterium rhinotracheale vaccination of broiler breeders on antibody titres and performance of breeders and broilers was investigated. O. rhinotracheale antibody titres and performance data were recorded from 16 different broiler breeder flocks and from 79 of their broiler progeny flocks. Eight breeder flocks were vaccinated with an inactivated O. rhinotracheale vaccine while the other eight breeder flocks were left unvaccinated against this bacterium. Following vaccination, mean O. rhinotracheale antibody titres in the breeders rose to a 6.5 log(2) units higher value than in unvaccinated breeders, and remained at a mean titre of 15 log(2) units during the entire production period. This resulted in significantly higher maternal antibody titres against O. rhinotracheale in the broiler progeny of vaccinated breeder flocks compared with the offspring of unvaccinated flocks. Statistical analyses revealed no differences in performance between vaccinated and unvaccinated breeders. There was a significantly lower mean mortality rate and higher mean production index in the broilers derived from vaccinated breeders.

Infectious bronchitis serology in broilers and broiler breeders: correlations between antibody titers and performance in vaccinated flocks.

In this study, a follow-up was made between 1993 and 1997 from broiler breeders at birth down to offspring broilers at processing, through vertically integrated registration of infectious bronchitis virus (IBV) antibody titers and performance data. All measurements were used two by two in a simple correlation study to calculate the degree to which they were linearly correlated. The antibody patterns in the broiler breeders indicated frequent field infections breaking through vaccinal immunity. Significant correlations measured between antibody titers and production parameters within and between the generations strongly suggested negative effects of IBV infections on laying percentage in the breeders and on mortality and daily weight gain in the broilers. Economic losses associated with IBV infections in the broilers occurred predominantly in flocks hatched with low and erratic maternal antibody titers. We concluded that IBV vaccination strategies should aim at high and uniform antibody titers in the broiler breeders.

Epidemiology and significance of chicken infectious anemia virus infections in broilers and broiler parents under nonvaccinated European circumstances.

A serologic survey in unvaccinated broiler parent and broiler progeny flocks demonstrated seroconversion against chicken infecrious anemia virus (CIAV) in all parent flocks before or around point of lay and in 38% of the broiler flocks examined at slaughter age. The presence of CIAV antibodies at slaughter of broilers was positively correlated with slaughterhouse condemnation rates. Results indicate that CIAV infections are highly prevalent in both broiler parent and broiler flocks and that CIAV infections in broilers are associated with increased slaughterhouse condemnation.

Secreted antigens as virulence-associated markers in Staphylococcus aureus strains from rabbits.

Western blot analysis was performed from the culture supernatant of 59 rabbit Staphylococcus aureus strains, classified as high and low virulence strains according to their epidemiological behaviour in commercial rabbitries, bio-, phage- and RAPD-type. Fourteen extracellular antigen bands (A-N) were recognised using sera of rabbits immunised with washed, viable high virulence S. aureus bacteria. Eleven of these bands were found in high virulence as well as in low virulence strains. The band A, approximately 78 kDa, was not seen in any of the 27 high virulence strains, except for one strain which was also typical in other aspects, was detected in all, but one of the low virulence strains. The M and N bands with molecular masses of approximately 29 and 27 kDa, respectively, were recognised in all high virulence strains except for the atypical strain, but in none of the low virulence strains. This indicates that the latter two antigens may be virulence-associated markers for S. aureus strains from rabbits.

Sequence analysis of a RAPD band differentiating high and low virulence Staphylococcus aureus strains from rabbits.

RAPD typing revealed the presence of a nucleotide band in typical high virulence rabbit Staphylococcus aureus strains which was absent in low virulence strains and in an atypical high virulence strain. The nucleotide sequence of this band was determined. Primers within this sequence were developed and PCR products of eight typical high virulence, one atypical high virulence and nine low virulence rabbit S. aureus strains were sequenced. All low virulence strains and the atypical high virulence strain revealed a constant difference with the typical high virulence strains for nucleotide 377 of the 1055bp sequence. The eight typical high virulence strains possessed a guanine base on this site, while the other strains tested showed an adenine base. These findings support the hypothesis on the clonal origin of typical high virulence rabbit S. aureus strains. After comparison with databases, two open reading frames (ORF) were identified within the sequence, which appeared to encode two structural ribosomal proteins. The single nucleotide mutation does not affect the amino acid sequence of the protein it encodes for.

Plasma biochemistry in pigeons experimentally infected with Salmonella.

Ten pigeons were crop inoculated with 1 x 10(9) colony-forming units of Salmonella typhimurium var. Copenhagen and observed during 28 days. Ten sham-inoculated pigeons served as noninfected controls. Clinical signs after Salmonella infection consisted of polydipsia, polyuria, and diarrhea. Morbidity was 90%, but there was no mortality. All inoculated pigeons showed fecal excretion of Salmonella for at least 7 days. Biochemical analysis of plasma samples taken at 3-day intervals indicated decreased concentrations of creatine kinase (CK)-MM and CK-MB isoenzymes and elevated total protein and alpha- and gamma-globulin values. No consistent changes in the level of 17 other blood parameters were observed. After 28 days, all pigeons were necropsied. Gross lesions and bacteriologic and histologic examination indicated septicemia in all Salmonella-inoculated pigeons. Results indicate that Salmonella septicemia in pigeons induces only limited changes in biochemical blood parameters. Decreased CK concentration was a consistent finding, however, and may therefore be a useful aid in the diagnosis of salmonellosis in pigeons.

Induction of the respiratory burst in turtle peritoneal macrophages by Salmonella muenchen.

Peritoneal macrophages were collected from juvenile turtles 72h after intraperitoneal inoculation with a 3% Sephadex suspension. The macrophages were assayed for their chemiluminescent (CL) properties, reflecting their respiratory burst activity, after stimulation with Zymosan A, phorbol 12-myristate 13-acetate (PMA), N-formyl-methionyl-leucyl-phenylalanine (fMLP), and calcium ionophore A23187. Except for fMLP, all triggering agents induced a marked CL response. Luminol was used as the chemiluminescent probe. When comparing CL responses in temperatures ranging from 15 to 35 degrees C, lower assay temperatures induced lower and slower CL responses. Stimulation with viable Salmonella muenchen resulted in a distinct response. Bacteria, inactivated by means of heat or acetone, induced a faster and stronger oxidative burst. Opsonization of either viable or heat-inactivated S. muenchen with non-inactivated anti-S. muenchen serum, prepared in turtles, induced faster and higher CL responses. On the other hand, opsonization of acetone-inactivated S. muenchen caused CL responses to be slower and weaker. S. muenchen, opsonized with heat-inactivated turtle anti S. muenchen serum, induced higher responses than non-opsonized bacteria, but slower and weaker responses than bacteria opsonized with native turtle antiserum. No response was recorded after stimulation with LPS and the supernatant of heat-inactivated bacteria.

Antibiotic sensitivity and resistance in Ornithobacterium rhinotracheale strains from Belgian broiler chickens.

Establishing the antibiotic sensitivity of the avian respiratory pathogen Ornithobacterium rhinotracheale is difficult because of the organism's complex growth requirements and the unusually frequent occurrence of resistance. The minimal inhibitory concentrations of 10 antibiotics were determined for 45 strains of O. rhinotracheale from Belgian broiler chickens collected from 45 farms between 1995 and 1998. They were compared with the type strain, which was isolated from a turkey, and a strain isolated from a rook. All the broiler strains were resistant to lincomycin and to the beta-lactams ampicillin and ceftiofur. Less than 10% of the strains were sensitive to the macrolides tylosin and spiramycin, tilmicosin and flumequine. A few strains were sensitive to enrofloxacin and doxycycline. All strains were sensitive to tiamulin.

Significance of infectious bursal disease serology in an integrated quality control program under European epidemiologic conditions.

In this study performed between 1993 and 1997, infectious bursal disease virus (IBDV) antibody titers and performance data were recorded in a vertically integrated monitoring scheme in order to make a follow-up from day-old parents down to the broilers at slaughter. All measured data were used two by two in a simple correlation study to calculate the degree to which they were linearly correlated. It appeared that high and/or uniform antibody titers in the parents were correlated with increased daily weight gain and decreased mortality and slaughterhouse condemnation in the broilers. Antibody titers and their CVs were negatively correlated in broiler parents and their offspring at day-old and even at slaughter. Results indicate that high and uniform antibody titers against IBDV in broiler parents are important for good performance of the broiler offspring, at least under the epidemiologic conditions of this study, which included the presence of very virulent IBDV strains in the field and the sole use of live intermediate vaccines in broilers as well as broiler parents.

Adhesion of Streptococcus gallolyticus strains to extracellular matrix proteins.

Fourteen pigeon Streptococcus gallolyticus strains of differing virulence, were tested for their ability to adhere to immobilised fibronectin, collagen types I, III and IV. Eight, 2 and 13 strains were able to bind fibronectin, collagen types III and IV, respectively. None of the strains adhered to collagen type I. Heat treatment, proteolytic digestion or periodate treatment reduced the binding of S. gallolyticus to fibronectin and collagen type IV, suggesting that surface receptors contain proteins and carbohydrates. Although binding to these extracellular matrix proteins can play a role in the pathogenesis of streptococcosis in pigeons, binding properties could not be related to virulence, indicating that other factors determine differences in virulence among pigeon S. gallolyticus strains. Adhesion to collagen type IV may account in part for the distribution pattern of the lesions observed in naturally and experimentally infected pigeons.