The expression of GapA and CrmA correlates with the Mycoplasma gallisepticum in vitro infection process in chicken TOCs.

Mycoplasma (M.) gallisepticum is the most pathogenic mycoplasma species in poultry. Infections cause mild to severe clinical symptoms associated with respiratory epithelial lesion development. Adherence, biofilm formation, and cell invasion of M. gallisepticum contribute to successful infection, immune evasion, and survival within the host. The important M. gallisepticum membrane-bound proteins, GapA and CrmA, are key factors for host cell interaction and the bacterial life-cycle, including its gliding motility, although their precise role in the individual infection step is not yet fully understood. In this study, we investigated the correlation between the host-pathogen interaction and the GapA/CrmA expression in an environment that represents the natural host's multicellular compartment. We used an in vitro tracheal organ culture (TOC) model, allowing the investigation of the M. gallisepticum variants, Rlow, RCL1, RCL2, and Rhigh, under standardised conditions. In this regard, we examined the bacterial adherence, motility and colonisation pattern, host lesion development and alterations of mucociliary clearance. Compared to low virulent RCL2 and Rhigh, the high virulent Rlow and RCL1 were more efficient in adhering to TOCs and epithelium colonisation, including faster movement from the cilia tips to the apical membrane and subsequent cell invasion. RCL2 and Rhigh showed a more localised invasion pattern, accompanied by significantly fewer lesions than Rlow and RCL1. Unrelated to virulence, comparable mucus production was observed in all M. gallisepticum infected TOCs. Overall, the present study demonstrates the role of GapA/CrmA in virulence factors from adherence to colonisation, as well as the onset and severity of lesion development in the tracheal epithelium.

Influence of heat stress on intestinal integrity and the caecal microbiota during Enterococcus cecorum infection in broilers.

Enterococcus cecorum (EC) is one of the most relevant bacterial pathogens in modern broiler chicken production from an economic and animal welfare perspective. Although EC pathogenesis is generally well described, predisposing factors are still unknown. This study aimed to understand the effect of heat stress on the caecal microbiota, intestinal integrity, and EC pathogenesis. A total of 373 1-day-old commercial broiler chicks were randomly assigned to four groups: (1) noninoculated, thermoneutral conditions (TN); (2) noninoculated, heat stress conditions (HS); (3) EC-inoculated, thermoneutral conditions (TN + EC); and (4) EC-inoculated, heat stress conditions (HS + EC). Birds were monitored daily for clinical signs. Necropsy of 20 broilers per group was performed at 7, 14, 21, and 42 days post-hatch (dph). A trend towards enhanced and more pronounced clinical disease was observed in the EC-inoculated, heat-stressed group. EC detection rates in extraintestinal tissues via culture were higher in the HS + EC group (~19%) than in the TN + EC group (~11%). Significantly more birds were colonized by EC at 7 dph in the HS + EC group (100%) than in the TN + EC group (65%, p < 0.05). The caecal microbiota in the two EC-inoculated groups was significantly more diverse than that in the TN group (p < 0.05) at 14 dph, which may indicate an effect of EC infection. An influence of heat stress on mRNA expression of tight junction proteins in the caecum was detected at 7 dph, where all six investigated tight junction proteins were expressed at significantly lower levels in the heat stressed groups compared to the thermoneutral groups. These observations suggest that heat stress may predispose broilers to EC-associated disease and increase the severity thereof. Furthermore, heat stress may impair intestinal integrity and promote EC translocation.

Infectious bursal disease virus' interferences with host immune cells: what do we know?

Infectious bursal disease virus (IBDV) induces one of the most important immunosuppressive diseases in chickens leading to high economic losses due to increased mortality and condemnation rates, secondary infections and the need for antibiotic treatment. Over 400 publications have been listed on PubMed.gov in the last 5 years pointing out the research interest in this disease and the development of improved preventive measures. While B cells are the main target cells of the virus, other immune and non-immune cell populations are also affected, leading to a multifaceted impact on the normally well-orchestrated immune system in IBDV-infected birds. Recent studies clearly revealed the contribution of innate immune cells as well as T cells to a cytokine storm and subsequent death of affected birds in the acute phase of the disease. Transcriptomics identified differential regulation of immune-related genes between different chicken genotypes as well as virus strains, which may be associated with a variable disease outcome. The recent availability of primary B cell culture systems allowed a closer look into virus-host interactions during IBDV infection. The new emerging field of research with transgenic chickens will also open up new opportunities to understand the impact of IBDV on the host under in vivo conditions, which will help to understand the complex virus-host interactions further.

Tenacity of Enterococcus cecorum at different environmental conditions.

AIMS: Our aim was to analyse the survival of Enterococcus cecorum (EC) at various temperatures, relative air humidities and on different substrates commonly existing in broiler houses. METHODS AND RESULTS: A pathogenic EC isolate (EC14) was used to inoculate sterile litter, polyvinyl chloride (PVC) and dust samples. Incubation at 37, 25 or 15°C with either 32% relative humidity (RH) or 78% RH followed. At defined time points (0-4272 h post-inoculation), samples were examined in triplicate for the total viable count. Selected combinations were repeated for a non-pathogenic and two additional pathogenic EC strains. For EC14, the measured survival time ranged from 48 to 4272 h (178 days) depending on the substrate-humidity-temperature combination. The longevity was the highest on litter, followed by dust and then PVC. Lower temperatures facilitated its survival, lower relative air humidity favoured the survival only in combination with 25 or 15°C. All three pathogenic strains showed longer survival times (up to 432 h, 18 days) compared to the non-pathogenic EC strain (168 h, 7 days) under the same conditions. CONCLUSIONS: Enterococcus cecorum demonstrates a high persistence in the environment especially at 15°C and 32% RH. SIGNIFICANCE AND IMPACT OF THE STUDY: Hygiene management plans should consider the durability of EC and the risk of a carry-over to control consecutive EC outbreaks.

Development of an in-house ELISA for detection of antibodies against Enterococcus cecorum in Pekin ducks.

Enterococcus cecorum (EC) is known to cause skeletal lesions in broiler chickens and also systemic infections in Pekin ducks. Despite the importance of the pathogen, there is still a lack of serological diagnostic tools for the detection of EC infections. Here we describe the development of an in-house indirect enzyme-linked immunosorbent assay (ELISA) for the detection of EC-specific antibodies and its application by examination of 67 sera from experimentally infected Pekin ducks, 710 field samples from four Pekin duck breeder flocks previously vaccinated with inactivated vaccines, and 80 samples from commercial Pekin ducks coming from vaccinated parent flocks. All groups that had been experimentally inoculated via the air sac route were positive in the new ELISA, with significantly (P ≤ 0.05) increased mean sample/positive (S/P) ratios of 0.71-2.70 at days 7, 14 and 21 post-infection, while orally inoculated ducks and the EC-free control group remained negative with mean S/P ratios of 0.0-0.15. Antibodies were also detected in each of four vaccinated Pekin duck breeder flocks; 67.8% of the samples were antibody positive. The highest S/P ratios were found between 16 and 26 weeks (median S/P ratios from 0.15 to 1.03), but antibodies were still detected in some serum samples in weeks 61-67 post-hatch. No antibodies were detected in the commercial Pekin ducks. Antibody development in the ducks may be influenced by the composition of the inactivated vaccine. The new ELISA provides a useful tool for investigations of response to EC infections and vaccinations.

In vitro characterization and genetic diversity of Bordetella avium field strains.

Bordetella avium (BA) is a respiratory pathogen of particular importance for turkeys. Specific adherence and damage to the respiratory epithelia are crucial steps of the pathogenesis, but knowledge about the mechanisms and the variety of virulence in field strains is limited. We analysed 17 BA field strains regarding their in vitro virulence-associated properties in tracheal organ cultures (TOC) of turkey embryos, and their genetic diversity. The TOC adherence assay indicated that BA field strains differ considerably in their ability to adhere to the tracheal mucosa, while the TOC ciliostasis assay illustrated a high degree of diversity in ciliostatic effects. These two virulence-associated properties were associated with each other in the investigated strains. Three of the investigated strains displayed significantly (P > 0.05) lower in vitro virulence in comparison to other strains. Genetic diversity of BA strains was analysed by core genome multilocus sequence typing (cgMLST). We applied a cgMLST scheme comprising 2667 targets of the reference genome (77.3% of complete genome, BA strain 197N). The results showed a broad genetic diversity in BA field strains but did not demonstrate a correlation between sequence type and virulence-associated properties. The cgMLST analysis revealed that strains with less marked virulence-associated properties had a variety of mutations in the putative filamentous haemagglutinin gene. Likewise, amino acid sequence alignment indicated variations in the protein. The results from our study showed that both adherence and ciliostasis assay can be used for virulence characterization of BA. Variations in the filamentous haemagglutinin protein may be responsible for reduced virulence of BA field strains.

Valid publication of the names Caecibacterium and Caecibacterium sporoformans.

Descriptions of the genus Caecibacterium and its proposed type species Caecibacterium sporoformans were published in the IJSEM by Onrust et al. (Int J Syst Evol Microbiol 2017;67:4589-4594). The type strain was deposited as LMG 27730 and DSM 26959. DSM 26959 is a patent strain, and therefore the names were effectively, but not validly, published based on Rule 30(4) of the International Code of Nomenclature of Prokaryotes. The type strain of C. sporoformans is now available from the Deutsche Sammlung von Mikroorganismen und Zellkulturen as DSM 103070 and no restrictions have been placed on its distribution. We here present new descriptions of the genus and its type species so that the names can be validly published.

Establishment of a Bordetella avium challenge model in turkeys.

Despite the importance of Bordetella avium (BA) as a respiratory pathogen of young turkeys, no infection model for the evaluation of BA-vaccine efficacy is available. The objective of this study was to evaluate the influence of route and dose of infection on the establishment of a BA-challenge model. In our first experiment, 28-day-old turkeys were either inoculated oculonasally with 105, 107 or 109 colony forming units (CFU) of BA per bird or exposed to BA by aerosol with 105-108 CFU/m3. The respiratory tract of all inoculated birds was BA-colonized, which was confirmed by choanal swabs and samples of trachea and lung, showing the highest prevalence in the aerosol-inoculated group. BA-specific humoral immune response was detected in the form of IgG in serum from five days post infection (dpi) and IgA in lacrimal fluid from seven dpi. In the second experiment, the model was tested in a vaccination trial. Twenty-one-day-old turkeys were vaccinated with a formalin-inactivated BA vaccine intramuscularly and challenged 21 days post vaccination with 107 CFU per bird oculonasally. BA-specific IgG antibodies were detected in serum and in lacrimal fluid 14 days post vaccination. As in the first experiment, secretory BA-specific antibodies of the IgA isotype were only detected in the inoculated groups from seven dpi. Despite the lack of clinical signs or pathological alterations in both experiments, vaccine efficacy was demonstrated by significant reduction in BA colonization of the trachea (P ≤ 0.05). In our study, a reliable model for BA infection has been established and has been demonstrated to be suitable for evaluation of vaccine efficacy.

Influence of the Gut Microbiota Composition on Campylobacter jejuni Colonization in Chickens.

The Campylobacter jejuni-host interaction may be affected by the host's gut microbiota through competitive exclusion, metabolites, or modification of the immune response. To understand this interaction, C. jejuni colonization and local immune responses were compared in chickens with different gut microbiota compositions. Birds were treated with an antibiotic cocktail (AT) (experiments 1 and 2) or raised under germfree (GF) conditions (experiment 3). At 18 days posthatch (dph), they were orally inoculated either with 104 CFU of C. jejuni or with diluent. Cecal as well as systemic C. jejuni colonization, T- and B-cell numbers in the gut, and gut-associated tissue were compared between the different groups. Significantly higher numbers of CFU of C. jejuni were detected in the cecal contents of AT and GF birds, with higher colonization rates in spleen, liver, and ileum, than in birds with a conventional gut microbiota (P < 0.05). Significant upregulation of T and B lymphocyte numbers was detected in cecum, cecal tonsils, and bursa of Fabricius of AT or GF birds after C. jejuni inoculation compared to the respective controls (P < 0.05). This difference was less clear in birds with a conventional gut microbiota. Histopathological gut lesions were observed only in C. jejuni-inoculated AT and GF birds but not in microbiota-colonized C. jejuni-inoculated hatchmates. These results demonstrate that the gut microbiota may contribute to the control of C. jejuni colonization and prevent lesion development. Further studies are needed to identify key players of the gut microbiota and the mechanisms behind their protective role.

Colonization patterns of Enterococcus cecorum in two different broiler production cycles detected with a newly developed quantitative real-time PCR.

BACKGROUND: Although Enterococcus cecorum (EC) infection is one of the most important bacterial diseases in modern broiler chickens today, many aspects of epidemiology and pathogenesis are still unknown. There is a need for better detection methods for EC than classical cultivation. In the present study, we describe the validation and application of a newly developed quantitative TaqMan real-time PCR (qPCR) assay based on the 16S-rRNA-gene for the detection of EC. RESULTS: Fifty EC strains isolated from 12 different animal species were detected with the assay, while none of the other 26 examined bacterial species were tested positive during validation procedure. The detection limit of the PCR was 6.25 CFU/ml PBS. The qPCR assay was also considerably more sensitive using intestine and organ samples than the classical cultivation method. Field application of the PCR setup was tested comparing two different broiler production cycles on one farm: in cycle I broilers showed signs of enterococcal spondylitis (ES) from day 24 post hatch onwards while broilers in cycle II developed no ES. Two totally different colonization patterns were found in the two cycles with the qPCR using cloacal swabs. Animals in cycle I showed significantly (P ≤ 0.05) higher detection rates of EC at the day of placement and throughout the cycle than broilers of cycle II. Additionally, significantly higher detection rates were found in the cecum compared to duodenum, jejunum and ileum within one cycle. CONCLUSIONS: The new qPCR for EC is highly specific, more sensitive than classical cultivation and was able to show differences in colonization in a broiler cycle with later EC disease outbreak compared to a healthy cycle. These findings may be explained by infection with different strains, pathogenic EC isolates are probably more effective in colonization than commensal isolates. A high correlation was found between qPCR results from cecum and cloacal swabs in this study, indicating that cloacal swabs can be used to examine intestinal colonization of broilers with EC. The new qPCR significantly improves the diagnostic of EC infections and may help to answer open questions concerning epidemiology and pathogenesis.

Caecibacterium sporoformans gen. nov., sp. nov., an anaerobic, butyrate-producing, spore-forming bacterium isolated from chicken caecum.

Strains of a Gram-stain-negative, rod-shaped and immotile bacterium were isolated from broiler chicken caecal content. The isolates required strict anaerobic conditions for growth, formed spores, were catalase-positive and oxidase-negative. They produced butyrate as the major metabolic end product in reinforced clostridial medium broth. The genomic DNA G+C content of the isolated strains was 32.5-34.6 mol%. The major cellular fatty acids were C16 : 0 FAME, C14 : 0 FAME, C19 : 0CYC 9,10DMA and C16 : 0DMA. The fatty acid composition of the cell wall showed no similarity to any strain in the midi database. 16S rRNA gene sequence analysis showed that the nearest phylogenetic neighbours were Anaerostipes hadrus and Clostridium populeti (92 % sequence similarity) within Clostridium cluster XIVa of the phylum Firmicutes. Therefore, a novel genus is proposed, with the name Caecibacterium sporoformans gen. nov., sp. nov. The type strain of Caecibacterium sporoformans is LMG 27730T=DSM 26959T.

Mycoplasma gallisepticum modifies the pathogenesis of influenza A virus in the avian tracheal epithelium.

Multiple respiratory infections have a significant impact on health and economy. Pathogenesis of co-infecting viruses and bacteria and their interaction with mucosal surfaces are poorly characterized. In this study we established a co-infection model based on pre-incubation of tracheal organ cultures (TOC) with Mycoplasma (M.) gallisepticum and a subsequent infection with avian influenza virus (AIV). Mycoplasma gallisepticum modified the pathogenesis of AIV as demonstrated in TOC of two different avian species (chickens and turkeys). Co-infection promoted bacterial growth in tracheal epithelium. Depending on the interaction time of M. gallisepticum with the host cells, AIV replication was either promoted or suppressed. M. gallisepticum inhibited the antiviral gene expression and affected AIV attachment to the host cell by desialylation of α-2,3 linked sialic acids. Ultrastructural analysis of co-infected TOC suggests that both pathogens may attach to and possibly infect the same epithelial cell. The obtained results contribute to better understanding of the interaction dynamics between M. gallisepticum and AIV. They highlight the importance of the time interval between infections as well as the biological properties of the involved pathogens as influencing factors in the outcome of respiratory infections.

Harryflintia acetispora gen. nov., sp. nov., isolated from chicken caecum.

More than 100 isolates of a Gram-stain-negative, curved rod-shaped, so far uncultured bacterium were isolated from chicken caecal content. The strains were obligate anaerobes which formed endospores. They were catalase-positive, cytochrome-oxidase-negative and did not hydrolyse aesculin. The detected predominant metabolic end product in reinforced clostridial bedium broth was acetate. The DNA G+C content was 62.2 mol%. The cell fatty acid composition was dominated by C15 : 0 iso, C15 : 0anteiso, C16 : 0, C13 : 0 iso, C14 : 0 iso and C16 : 0DMA. Analysis of the 16S rRNA gene sequences of the novel isolates showed 99 % similarity to each other. Phylogenetically they are placed in Clostridium cluster IV. They exhibited 99 % 16S rRNA gene sequence similarity to uncultured bacterial clone SJTU_C_03_83 (GenBank accession no. EF404038.1) and to three other so far uncultured strains. The closest related type strains, Hydrogenoanaerobacterium saccharovorans SW51T and Anaerotruncus colihominis WAL 14565T, exhibited a 16S rRNA gene sequence divergence of at least 8 % to the newly described isolates. In addition, morphological and biochemical characteristics allowed differentiation of the novel isolates from these and other related and validly published species, including Clostridium methylpentosum and Acetanaerobacterium elongatum. Therefore, a new genus, Harryflintia gen. nov., is proposed for the isolated hitherto unknown strains with V20-281aT (=DSM 100433T =CECT 8892T) as the type strain of its type species, Harryflintia acetispora gen. nov., sp. nov.

Comparison of infectious bursal disease live vaccines and a HVT-IBD vector vaccine and their effects on the immune system of commercial layer pullets.

Infectious bursal disease (IBD) is an economically important disease affecting poultry production worldwide. Previous experimental studies indicated that IBD live vaccination may induce transient immunosuppression, leading to suboptimal vaccine responses and therefore insufficient protection against other pathogens. Layer pullets are commonly not only vaccinated against IBD within their rearing period, but also against a variety of other pathogens. Therefore, it is of interest to investigate the effects of different IBD vaccination regimes on conventionally applied vaccines against other pathogens, and possible protection against widely spread very virulent IBD-virus (vvIBDV). A commercially available Herpesvirus of turkey vector vaccine (vHVT-IBD) expressing viral protein 2 of IBDV, and two IBD live vaccines were compared in commercial pullets for their effects on circulating B cell numbers, the ability of vaccinated birds to mount a humoral immune response against different antigens as well as their ability to induce protection against vvIBDV challenge. The results of this study demonstrate a clear immunosuppressive effect of the intermediate plus IBD live vaccine on the humoral branch of the immune system. On the other hand, no detectable effects of vHVT-IBD vaccination on these parameters were observed. All tested IBD vaccines protected against clinical IBD, although none induced sterile immunity in commercial layer pullets. vHVT-IBD-vaccinated birds showed significantly less lesions after vvIBDV challenge than IBD live-vaccinated or non-vaccinated birds (P < 0.05). Therefore, vHVT-IBD may be a suitable alternative to conventional IBD live vaccines, and may be applied even in the presence of maternally derived IBD antibodies without induction of detectable humoral immunosuppression.

Avian metapneumovirus infection of chicken and turkey tracheal organ cultures: comparison of virus-host interactions.

Avian metapneumovirus (aMPV) is a pathogen with worldwide distribution, which can cause high economic losses in infected poultry. aMPV mainly causes infection of the upper respiratory tract in both chickens and turkeys, although turkeys seem to be more susceptible. Little is known about virus-host interactions at epithelial surfaces after aMPV infection. Tracheal organ cultures (TOC) are a suitable model to investigate virus-host interaction in the respiratory epithelium. Therefore, we investigated virus replication rates and lesion development in chicken and turkey TOC after infection with a virulent aMPV subtype A strain. Aspects of the innate immune response, such as interferon-α and inducible nitric oxide synthase mRNA expression, as well as virus-induced apoptosis were determined. The aMPV-replication rate was higher in turkey (TTOC) compared to chicken TOC (CTOC) (P < 0.05), providing circumstantial evidence that indeed turkeys may be more susceptible. The interferon-α response was down-regulated from 2 to 144 hours post infection in both species compared to virus-free controls (P < 0.05); this was more significant for CTOC than TTOC. Inducible nitric oxide synthase expression was significantly up-regulated in aMPV-A-infected TTOC and CTOC compared to virus-free controls (P < 0.05). However, the results suggest that NO may play a different role in aMPV pathogenesis between turkeys and chickens as indicated by differences in apoptosis rate and lesion development between species. Overall, our study reveals differences in innate immune response regulation and therefore may explain differences in aMPV - A replication rates between infected TTOC and CTOC, which subsequently lead to more severe clinical signs and a higher rate of secondary infections in turkeys.

Co-infection with Multiple Respiratory Pathogens Contributes to Increased Mortality Rates in Algerian Poultry Flocks.

Respiratory infections are a common cause for increased mortality rates in poultry worldwide. To improve intervention strategies, circulating pathogens have to be identified and further characterized. Because of the lack of diagnostic tools, it was not known what pathogens contribute to the high mortality rates in association with respiratory disease in Algeria. Our objective was to determine if primary pathogens including Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS), avian influenza virus (AIV), infectious bronchitis virus (IBV), and avian metapneumovirus (aMPV), known to be present in neighboring countries, can also be detected in Algerian chicken and turkey flocks. Results demonstrate the circulation of the investigated pathogens in Algerian poultry flocks as multi-infections. Phylogenetic characterization of the Algerian IBV strains confirmed the circulation of nephropathogenic viruses that are different from the strains isolated in neighboring countries. This could suggest the existence of a new IBV genotype in North Africa. Additionally, we detected for the first time an aMPV subtype B field strain and avian influenza virus. Interestingly, all viral pathogens were present in co-infections with MG, which could exacerbate clinical disease. Additional pathogens may be present and should be investigated in the future. Our results suggest that multiple respiratory infections may be responsible for high mortality in Algerian poultry flocks and very probably also in other regions of the world, which demonstrates the need for the establishment of more comprehensive control strategies.

Reassortment of NS segments modifies highly pathogenic avian influenza virus interaction with avian hosts and host cells.

Highly pathogenic avian influenza viruses (HPAIV) of subtypes H5 and H7 have caused numerous outbreaks in diverse poultry species and rising numbers of human infections. Both HPAIV subtypes support a growing concern of a pandemic outbreak, specifically via the avian-human link. Natural reassortment of both HPAIV subtypes is a possible event with unpredictable outcome for virulence and host specificity of the progeny virus for avian and mammalian species. NS reassortment of H5N1 HPAIV viruses in the background of A/FPV/Rostock/1934 (H7N1) HPAIV has been shown to change virus replication kinetics and host cell responses in mammalian cells. However, not much is known about the virus-host interaction of such viruses in avian species. In the present study, we show that the NS segment of A/Vietnam/1203/2004 (FPV NS VN, H5N1) HPAIV significantly altered the characteristics of the H7 prototype HPAIV in tracheal organ cultures (TOC) of chicken and turkey in vitro, with decreased replication efficiency accompanied by increased induction of type I interferon (IFN) and apoptosis. Furthermore, species-specific differences between chicken and turkey were demonstrated. Interestingly, NS-reassortant FPV NS VN showed an overall highly pathogenic phenotype, with increased virulence and replication potential compared to the wild-type virus after systemic infection of chicken and turkey embryos. Our data demonstrate that single reassortment of an H5-type NS into an H7-type HPAIV significantly changed virus replication abilities and influenced the avian host cell response without prior adaptation.

Differences in the tissue tropism to chicken oviduct epithelial cells between avian coronavirus IBV strains QX and B1648 are not related to the sialic acid binding properties of their spike proteins.

The avian coronavirus (AvCoV) infectious bronchitis virus (IBV) is a major poultry pathogen. A characteristic feature of IBV is the occurrence of many different strains belonging to different serotypes, which makes a complete control of the disease by vaccinations a challenging task. Reasons for differences in the tissue tropism and pathogenicity between IBV strains, e.g. a predilection for the kidneys or the oviduct are still an open question. Strains of the QX genotype have been major pathogens in poultry flocks in Asia, Europe and other parts of the world. They are the cause of severe problems with kidney disease and reproductive tract disorders. We analysed infectivity and binding properties of the QX strain and compared them with those of the nephropathogenic strain B1648. As most IBV strains do not infect permanent cell lines and show infection only in primary chicken cells of the target organs, we developed a culture system for chicken oviduct explants. The epithelial cells of the oviduct showed a high susceptibility to infection by the QX strain and were almost resistant to infection by the nephropathogenic B1648 strain. Binding tests with isolated primary oviduct epithelial cells and soluble S1 proteins revealed that S1 proteins of two IBV strains bound with the same efficiency to oviduct epithelial cells. This attachment was sialic acid dependent, indicating that the sugar binding property of IBV spike proteins is not the limiting factor for differences in infection efficiency for the oviduct of the corresponding viruses.

Comprehensive report of an Enterococcus cecorum infection in a broiler flock in Northern Germany.

BACKGROUND: Enterococcus cecorum is considered as an emerging pathogen in poultry and can cause substantial losses in broiler flocks. Femoral head necrosis and spondylitis were described as the main pathological changes in infected chickens. Nevertheless, little is known about the pathogenesis of Enterococcus cecorum infection in broilers. This report shows for the first time the whole course of disease over an entire growing period including repeated necropsies and subsequent microbiological investigations. CASE PRESENTATION: In a flock of 18200 broilers, a decrease in flock uniformity was detected from 14 days post hatch onwards with affected chickens showing lameness and an increase in flock mortality up to 7.22% at day 33 post hatch. In the first 3 weeks post hatch, pericarditis and hepatitis were found as the main pathological changes in 27.6% and 9.8% of the examined broilers respectively. Femoral head necrosis and vertebral osteomyelitis were detected in the last week of the growing period with 10.3% and 2.3% respectively. Heart, liver, spleen, yolk sac and vertebral column of 59 broilers with pathological changes were subjected to bacteriological analysis. Enterococcus cecorum was isolated from 23 birds (39%), the first broiler was already positive at day 3 post hatch in the yolk sac. Additionally, 9.75% of the broilers were rejected at the slaughterhouse primarily because of pathological changes. The investigated broiler cycle had by far the best footpad score compared to 7 cycles before and 4 cycles after the Enterococcus cecorum infection at the same farm. CONCLUSIONS: Bacteraemia and generalized infection appear to be important steps in the pathogenesis of Enterococcus cecorum infection in broilers. Furthermore, this disease causes economic losses for the farmer not only due to an increase in flock mortality, but probably also through substantially higher condemnation rates at the slaughterhouse. It was speculated that the broilers were infected via the respiratory tract as this flock had lower footpad scores likely the result of drier litter. The latter may have led to higher dust concentrations and thus airborne Enterococcus cecorum.

Enterococcus cecorum infection in a racing pigeon.

Until now, Enterococcus cecorum (EC) has been known as a pathogen for broilers, broiler breeders, and Pekin ducks. In the present report, we describe a fatal systemic EC infection in a young racing pigeon (Columba livia forma domestica). EC was isolated from the heart, liver, spleen, and intestine of the bird in pure culture. In the pathologic examination, the pigeon showed enteritis and an ulcerative gastritis, which may have been predisposing factors for the development of the generalized EC infection. An accumulation of gram-positive cocci in spleen tissue was found in the histopathologic examination and confirms the presence of a systemic EC infection in the pigeon. Additionally, EC was isolated from cloacal swabs of other pigeons in the same loft, but no additional pigeons were submitted for necropsy. All EC isolates tested were negative by PCR for the enterococcal virulence factors cytolysin, enterococcal surface protein, aggregation substance, hyaluronidase, and gelatinase. Therefore, the reason for the enhanced virulence of the EC isolate remains unknown. Our report confirms EC as a disease-causing agent in pigeons and presents the first data concerning the analysis of EC for virulence factors.