Indications for the use of highest priority critically important antimicrobials in the veterinary sector.

BACKGROUND: Among the measures taken to preserve the clinical efficacy of highest priority critically important antimicrobials (HP-CIAs), the WHO has recommended avoiding their use in food-producing animals. Little is known regarding the indications for which different antimicrobial classes are used in animals, even in countries where data on antimicrobial use are available. OBJECTIVES: To outline, in a narrative review, the diseases for which HP-CIAs are used in veterinary medicine, highlighting incongruences with international guidelines and disease conditions where effective alternatives to HP-CIAs are missing. METHODS: Scientific literature, national reports and expert opinion were used to describe the indications for the use of HP-CIAs in the main food-producing (pigs, cattle and poultry) and companion (horses, dogs and cats) animal species. RESULTS: The most common indications for use of HP-CIAs are enteric and respiratory infections in pigs, cattle and poultry, urogenital infections in dogs and cats and respiratory infections in horses. In some instances, no valid and convenient alternatives to colistin and macrolides are available against certain porcine enteric and bovine respiratory pathogens. Effective, legal and convenient alternatives to HP-CIAs are also lacking for managing common infections in cats, for which oral administration is difficult, Rhodococcus equi infections in horses, some enteric and respiratory infections in poultry and MDR infections in all companion animal species. CONCLUSIONS: Future research and stewardship programmes should focus on the disease conditions identified by this review to reduce the use of HP-CIAs in the veterinary sector.

Whole genome sequence comparison of avian pathogenic Escherichia coli from acute and chronic salpingitis of egg laying hens.

BACKGROUND: Infection in the oviduct (salpingitis) is the most common bacterial infection in egg laying hens and is mainly caused by Escherichia coli. The disease is responsible for decreased animal welfare, considerable economic loss as well as a risk of horizontal and vertical transmission of pathogenic E. coli. The outcome of salpingitis may be either acute or chronic. It has not yet been clarified whether the pathological manifestation is a result of the characteristics of the E. coli or whether the manifestation is associated with host factors such as host immunity. RESULTS: From the core- and accessory genome analysis and comparison of 62 E. coli no genetic markers were found to be associated to either acute or chronic infection. Twenty of the 62 genomes harboured at least one antimicrobial resistance gene with resistance against sulfonamides being the most common. The increased serum survival and iron chelating genes iss and iroN were highly prevalent in genomes from both acute and chronic salpingitis. CONCLUSION: Our analysis revealed that no genetic markers could differentiate the E. coli isolated from acute versus chronic salpingitis in egg laying hens. The difference in pathological outcome may be related to other factors such as immunological status, genetics and health of the host. These data indicate that salpingitis is another manifestation of colibacillosis.

Correlation between footpad lesions and systemic bacterial infections in broiler breeders.

Footpad lesions are an important factor in evaluation of animal welfare in broilers regulated by law; however, no legal requirements have been set for the parent birds. Nevertheless, the present study confirms that foot health in broiler breeders declines significantly with increasing age, thus potentially impairing the animal welfare due to pain and discomfort from footpad dermatitis. Furthermore, this is the first report demonstrating a correlation between the presence of footpad lesions and systemic bacterial infections with Gram-positive cocci in broiler breeder birds.

Evaluation of the efficacy of an autogenous Escherichia coli vaccine in broiler breeders.

In poultry production Escherichia coli autogenous vaccines are often used. However, the efficacy of autogenous E. coli vaccinations has not been evaluated experimentally in chickens after start of lay. The aim of the present study was to evaluate the protective effect of an autogenous E. coli vaccine in broiler breeders. Three groups of 28-week-old broiler breeders (unvaccinated, vaccinated once and twice, respectively) were challenged with a homologous E. coli strain (same strain as included in the vaccine) or a heterologous challenge strain in an experimental ascending model. The clinical outcome was most pronounced in the unvaccinated group; however, the vast majority of chickens in the vaccinated groups had severe pathological manifestations similar to findings in the unvaccinated group after challenge with a homologous as well as a heterologous E. coli strain. Although significant titre rises in IgY antibodies were observed in the twice vaccinated group, antibodies did not confer significant protection in terms of pathological impact. Neither could transfer of maternal-derived antibodies to offspring be demonstrated. In conclusion, with the use of the present model for ascending infection, significant protection of an autogenous E. coli vaccine against neither a homologous nor a heterologous E. coli challenge could not be documented.

Performance of a commercial Chicken-Ovo-transferrin-ELISA on the serum of brown layer chickens infected with Gallibacterium anatis and Streptococcus zooepidemicus.

To evaluate Ovo-transferrin (OTF), a positive acute-phase protein in chickens, as a diagnostic biomarker of selected bacterial infections we checked the performance of a commercial Chicken-OTF-ELISA (ICL, Inc., Portland, OR, USA) by analytical and overlap performances using two groups of serum samples obtained from 26 Gallibacterium anatis-infected and 20 Streptococcus zooepidemicus-infected brown layer chickens. In addition, sera from 14 apparently healthy and 19 negative control chickens were analysed in the Gallibacterium group whereas sera from 20 healthy and 11 negative control chickens from the Streptococcus group were analysed. All calibration curves revealed high coefficients of determination (≥ 0.97) between optical density (OD 450nm) and concentrations of OTF (mg/ml). OTF concentrations in high, medium and low pools (made of sera from a combination of infected and/or non-infected birds) were >6.4, >3.8 to <4.5 and <1.6 mg/ml in the Gallibacterium group, and >6.7, >3.5 to <3.7 and <1.1 mg/ml in the Streptococcus group, respectively. For each pool, low coefficients of intra-assay (7.8, 5.7 and 5.3) and inter-assay (15.8, 18.0 and 18.0) variations were obtained in the Gallibacterium study. In the Streptococcus study only the intra-assay variation was low (3.7, 3.8 and 6.2, respectively). The linearity check was acceptable demonstrating a straight line with slope and intercept, not deviating from one and zero, respectively, using the Gallibacterium sera, whereas the Streptococcus sera deviated from the linear line. Detection limits were low (Gallibacterium, 0.01 mg/ml; Streptococcus, 0.32 mg/ml). OTF concentrations (mean ± standard error of the mean) in overlap performances were elevated in the sera of infected chickens (Gallibacterium, 4.4 ± 0.3 mg/ml; Streptococcus, 3.2 ± 0.4 mg/ml) compared with negative controls (1.7 ± 0.1 mg/ml) (P < 0.05). In conclusion, the Chicken-OTF-ELISA can be used to measure reproducible serum OTF concentrations in brown layer chickens as a response to G. anatis infections, whereas an adjustment of dilution process is proposed to optimize to use in S. zooepidemicus-infected chickens.

Inflammation-induced haemostatic response in layer chickens infected with Streptococcus equi subsp. zooepidemicus as evaluated by fibrinogen, prothrombin time and thromboelastography.

Streptococcus zooepidemicus has recently been shown to be a severe pathogen in layer chickens, where it is able to cause serious lesions in the vascular system. To evaluate the haemostatic response, 10 layer chickens were inoculated intravenously with S. zooepidemicus. Four hypotheses were tested: that the infection-induced inflammation would increase the plasma fibrinogen (Fbg) concentration, would prolong the prothrombin time (PT) and would prompt hypercoagulability or hypocoagulability as assessed by whole-blood thromboelastography (TEG), and that a possible correlation would exist between one of the TEG values and Fbg/PT. Each parameter was measured at days 1, 3 and 6 post inoculation (p.i.), and compared with the values at day 0 from each individual bird and with values obtained from non-infected control chickens (n = 10). In the infected chickens, the mean (± standard error) of Fbg was higher at day 3 p.i. (9.4 ± 1.4 g/l) and day 6 p.i. (8.0 ± 0.7 g/l) and the PT was prolonged at day 6 p.i. (168.1 ± 21.0 sec) compared with the day 0 standards (2.6 ± 0.2 g/l and 104.6 ± 2.0 sec, respectively) (P < 0.05). The majority of infected chickens demonstrated a hypercoagulable TEG result with increased mean values of the clot formation rate (α-angle) and maximal amplitude (MA) of TEG tracing at day 3 p.i. (83.1 ± 0.7°, 83.8 ± 1.4 mm) and day 6 p.i. (84.0 ± 0.4°, 89.8 ± 1.0 mm) compared with the day 0 values (75.8 ± 2.2° and 66.9 ± 1.4 mm, respectively) (P < 0.05). In control birds, the means of Fbg (1.5 ± 0.1 g/l), PT (79.4 ± 6.4 sec), TEG-α (76.7 ± 1.5°) and TEG-MA (64.0 ± 2.3 mm) were lower at day 6 compared with values observed for the infected chickens (P < 0.05). A negative correlation coefficient (-0.71) was found between the clot formation time (TEG-K) and Fbg at day 1 in the control group (P = 0.02). In conclusion, infection with S. zooepidemicus following intravenous injection in layer chickens induced haemostatic alterations including hyperfibrinogenaemia, prolonged PT, and hypercoagulability as measured by increased TEG-α and TEG-MA.

Pathogenicity of wild-type and small-colony variants of Streptococcus equi subsp. zooepidemicus in layer chickens.

Wild-type (WT) and small-colony-variant (SCV) strains of Streptococcus equi subsp. zooepidemicus have recently been isolated from a layer flock in Denmark experiencing high mortality. To investigate the disease-causing potential of SCV compared with WT, a 2-week long infection study was performed in 45-week-old brown layer chickens. Four groups of 11 chickens each were inoculated with a WT or SCV strain by the intravenous or intra-tracheal route: WT-IV, SCV-IV or WT-IT, SCV-IT, respectively. Clinical signs were observed in most chickens in the WT-IV group (9/11). Mortality was observed in the SCV-IV (4/11) and WT-IV (2/11) groups. Ten chickens in the WT-IV and WT-IT groups, respectively, developed gross lesions including oophoritis/peritonitis, hepatitis and airsacculitis cervicalis. Bronchopneumonia was common in the SCV-IT group (6/11), and valvular endocarditis in the SCV-IV group (4/11). Histological lesions in liver tissue were frequently observed in the chickens of the SCV-IV group (9/11), followed by the WT-IT (7/11), WT-IV (6/11), and SCV-IT (2/11) groups. The lesions in the SCV-IV group were dominated by deposition of eosinophilic material with infiltration of inflammatory cells (6/9). Bacteriological re-isolation of either strain type was achieved from all chickens of the WT-IV and WT-IT groups, and from nine and seven out of 11 chickens for each of the SCV-IV and SCV-IT groups, respectively. In summary, we were able to reproduce clinical signs and lesions as observed during the natural outbreak, which included an overall initial onset in WT-infected chickens as opposed to a late onset and possible recurring infection seen in the SCV-infected chickens.

Genomic traits of Aeromonas veronii isolated from slaughtered Danish broilers.

Aeromonas veronii is a Gram-negative opportunistic bacterium found in fish, poultry and humans and has occasionally been associated with disease although not generally considered a poultry pathogen. A. veronii was recently isolated from both healthy and condemned broiler carcasses at a major Danish abattoir. In this study, we did a whole genomic analysis of 24A. veronii strains from the abattoir to determine their potential sources and relatedness as well as their pathogenic potential, antimicrobial resistance determinants and associated mobile elements. No strains were multi-drug resistant, but all strains carried the beta-lactam resistance genes cphA3 and blaOXA-12 without being phenotypically resistant to carbapenems. One strain carried an IncA plasmid with tet(A), tet(B) and tet(E) genes. A phylogenetic tree including public A. veronii sequences showed that our isolates were not clonal but were dispersed around the phylogenetic tree, suggesting a diffuse spread of A. veronii across human, aquatic and poultry samples. Strains carried different virulence factors known to be associated with pathogenesis and severity of disease in animals and humans, e.g. type II (aerolysin, amylases, proteases, and cytotoxic enterotoxin Act) and III secretion systems where the latter has been associated with mortality in hospitalized patients. Although our genomic analysis of A. veronii shows zoonotic potential, epidemiological studies of human gastro-enteritis cases of A. veronii associated with consumption of broiler meat are needed. It remains to be proven if A. veronii is a true poultry pathogen and part of the established microflora in abattoirs and the gut-intestinal microflora of poultry.

Keel bone fractures in Danish laying hens: Prevalence and risk factors.

Keel bone fractures (KBF) in commercial poultry production systems are a major welfare problem with possible economic consequences for the poultry industry. Recent investigations suggest that the overall situation may be worsening. Depending on the housing system, fracture prevalences exceeding 80% have been reported from different countries. No specific causes have yet been identified and this has consequently hampered risk factor identification. The objective of the current study was to investigate the prevalence of KBF in Danish layer hens and to identify risk factors in relation to KBF in all major productions systems, including parent stock production. For risk factor identification, production data from the included flocks was used. In total, 4794 birds from 40 flocks were investigated at end-of-lay. All birds were euthanized on farm and underwent inspection and palpation followed by necropsy. All observations were recorded and subsequently analysed using the SAS statistical software package. In flocks from non-caged systems, fracture prevalence in the range 53%-100%, was observed whereas the prevalence in flocks from enriched cages ranged between 50-98%. Furthermore, often multiple fractures (≥4) were observed in individual birds (range 5-81% of the birds with fractures) depending on the flock. The localization of the fractures at the distal end of the keel bone is highly consistent in all flocks (>96%). Macroscopically the fractures varied morphologically from an appearance with an almost total absence of callus, most frequently observed in caged birds, to large callus formations in and around the fracture lines, which was a typical finding in non-caged birds. Despite being housed under cage-free conditions, parent birds had significantly fewer fractures (all flocks were 60 weeks old) per bird, than other birds from cage-free systems. The body weight at end-of-lay had an effect on the risk of having fractures, heavy hens have significantly fewer fractures at end-of-lay. The older the hens were at onset of lay, the lower was the flock prevalence at end-of-lay. Additionally, the daily egg size at onset of lay was of importance for the risk of developing fractures, the production of heavier eggs initially, resulted in higher fracture prevalence at depopulation. The odds ratio of body weight, (+100 g) was 0.97, age at onset of lay (+1 week) was 0.87 and daily egg weight at onset (+1 gram) was 1.03. In conclusion, the study demonstrated a very high prevalence of KBF in hens from all production systems and identified hen size, age at onset of lay and daily egg weight at onset of lay to be major risk factors for development of KBF in the modern laying hen. Further research regarding this is warranted to strengthen the longevity and enhance the welfare of laying hens.

Keel bone fractures are more prevalent in White Leghorn hens than in Red Jungle fowl hens-A pilot study.

Fractures and deviations to the keel bone are common in commercial laying hens, with reported variations in occurrence across strains and breeds. The aetiology is not fully understood, however, modern genetics and selection for efficient egg production has been claimed to be important factors for the keel bone fractures. To explore this further, we investigated keel bones from two different breeds, representing different degrees of selection for egg production: Red jungle fowl (n = 82), and White Leghorn (n = 32), where the latter is a selected laying breed which is the origin for many modern laying hen hybrids. Keel bones from a total of 116 birds, 53 hens and 63 roosters, were examined by necropsy at 80 weeks of age. All birds were raised in modified aviaries in the same holding facility. Overall, 24.5% of the hens had one or more fractures to the keel, with a difference in the prevalence between hens from the two breeds (p<0.01): 10% (95% CI: 3.7-24%) in the Red Jungle fowl hens and 69% (95% CI: 37-90%) in the White Leghorn hens. No roosters, regardless of breed, had keel bone fractures. Mild to moderate keel bone deviations were present in 54% (95% CI: 25-80%) of the hens and 4.7% (95% CI: 0.5-30%) of the roosters, all White Leghorns.

Pathological characterization of keel bone fractures in laying hens does not support external trauma as the underlying cause.

Keel bone fractures in laying hens have been described with increasing prevalence from several countries over the last twenty years and are considered one of the greatest welfare problems to the layer industry. In Denmark we have observed fracture prevalence in the range of 53% to 100% in flocks from cage-free systems whereas flock prevalences in birds from enriched cages ranged between 50-98%. Previous research have speculated that the underlying reason for the development of keel bone fractures is trauma in relation to impact of the bird with furniture, other equipment etc. However, little evidence of this theory has been provided. Predisposing factors have also been suggested including genetics of the bird, lack of specific feedstuff components, high egg production, management factors and layer fatigue. This study has addressed the possible pathogenesis of these fractures by pathological characterization of fractures in birds from different production systems. More than 60 keel bones with fractures have been characterized histo-pathologically and by CT scan. This included an assessment of damage to muscles and soft tissues, the bone and the healing process including callus formation. This investigation has shown that high energy collisions cannot be responsible for the majority of fractures, located at the caudal tip of the keel bone, observed in laying birds as markers associated trauma were not observed in the majority of the cases just as few recognized healing processes were observed. These results suggest an alternative pathogenesis to trauma.

Comparison and assessment of necropsy lesions in end-of-lay laying hens from different housing systems in Denmark.

Apperantly healthy laying hens at the end of production (60 to 91 wk) were investigated for the occurrence of pathology and bacterial infections. In total, 7,477 hens from 15 flocks representing the following production systems: Enriched cages, barn housed layers, and organic/free range layers were necropsied. Indications of bacterial infection were investigated by bacteriological cultivation. The overall prevalence of lesions was 16.60%, including lesions of both infectious and non-infectious origin. The most prevalent lesions were bursitis presternalis (6.65%), reproductive tract lesions (e.g., salpingitis and/or peritonitis and/or oophoritis) (3.50%), serosal scarification (e.g., fibrotic adhesive peritonitis) 1.55%, and neoplasm 1.73%. Significant differences were observed between different production systems and/or flocks in the prevalence of reproductive tract lesions, bursitis presternalis, serosal scarification, skin infections, juvenile hens, and traumas/fractures. No significant difference was observed between different production systems in the prevalence of neoplasia, infection of septicemic etiology, and pododermatitis. In total, 3.4% of the hens were out of lay, with significantly higher rate in organic flocks. Infections of the reproductive tract were the most prevalent lesions with bacterial etiology in all productions systems. In total, 40% of the hens with lesions associated to the oviduct were out of lay and significant difference between production systems were observed. Escherichia coli was the most commonly isolated bacteria and in 90% of the cases they were isolated from the reproductive tract lesions. The second most prevalent bacteria was Gallibacteruim anatis. Significant difference in the prevalence of E. coli positive hens was observed between production systems (P < 0.05). In conclusion, the prevalence of reproductive tract lesions in apparently healthy end-of-lay laying was higher than indicated in previous reports. These findings support the previous suggestions that E. coli and G. anatis are the major pathogens causing reproductive tract lesions.

Explanations for keel bone fractures in laying hens: are there explanations in addition to elevated egg production?

The current article served to provide the most up-to-date information regarding the causes of keel bone fracture. Although elevated and sustained egg production is likely a major contributing factor toward fractures, new information resulting from the development of novel methodologies suggests complementary causes that should be investigated. We identified 4 broad areas that could explain variation and increased fractures independent of or complementing elevated and sustained egg production: the age at first egg, late ossification of the keel, predisposing bone diseases, and inactivity leading to poor bone health. We also specified several topics that future research should target, which include continued efforts to link egg production and bone health, examination of noncommercial aves and traditional breeds, manipulating of age at first egg, a detailed histological and structural analysis of the keel, assessment of prefracture bone condition, and the relationship between individual activity patterns and bone health.

Prevalence of Keel Bone Damage in Red Jungle Fowls (Gallus gallus)-A Pilot Study.

Keel bone damage (KBD) is a highly prevalent problem in commercial egg production. KBD consists of two different conditions affecting the keel: Keel bone deviation and keel bone fractures (KBF). Deviations are linked to pressure on the keel, e.g., from perching. The causative factors for KBF are not clear; however, selection for efficient egg production has been suggested as a major contributing factor. An important step to shed light on the role of selective breeding as an underlying cause of KBF in modern laying hens is to evaluate the keel bones of the ancestor, the red jungle fowl. To the authors' knowledge, this has never previously been published. The aim of this study was therefore to describe the prevalence of KBD in a study group of red jungle hens and roosters housed in an aviary system. The present study examined 29 red jungle fowls 112 weeks of age post-mortem; 12 hens and 17 roosters. Keel bones were evaluated by external palpation for deviations and fractures. Palpation was followed by autopsy. No fractures were detected in the 17 roosters; one had a very slight deviation. Of the 12 red jungle hens in this pilot study, one had a single fracture and 10 hens had a very slight deviation.

A Descriptive Study of Keel Bone Fractures in Hens and Roosters from Four Non-Commercial Laying Breeds Housed in Furnished Cages.

The presence of keel bone fractures (KBF) in laying hens has been documented and discussed by several authors, nevertheless the causative factors behind KBF remain uncertain. High prevalence of KBF have been reported in all commercial egg production systems, in different genetic lines and at different ages. Several of the proposed causal mechanisms behind KBF are linked to selection for efficient production. It is, therefore, of interest to explore whether less selected breeds have a lower occurrence of keel bone fractures compared to reports from highly selected, modern laying hen breeds. Thus, the aim of the current study was to investigate keel bones of hens from four non-commercial layer breeds. Birds were housed in furnished cages and keel bones examined at 30 and 63 weeks of age, using a portable X-ray equipment. The results from this descriptive study indicate a low prevalence of KBF at both ages in all four breeds, with only five KBF detected in 213 X-ray pictures taken from 126 birds. Of these, four of the KBF were observed in the most genetically selected breed, with an early onset of lay. None of the roosters examined exhibited KBF. The overall low numbers of KBF found indicate that genetic factors may be involved in KBF and, thus that selective breeding may help to reduce the susceptibility to KBF. Finally, this study highlights the importance of poultry conservation to secure genetic diversity, which may be an important resource in future selection schemes.

Longitudinal Study on Causes of Mortality in Danish Broiler Breeders.

Broiler production is highly dependent on good health in the parent flocks. The so-called normal mortality in these flocks remains to be addressed to further reduce mortality of the breeders and to improve the quality of broilers. The aim of the present study, therefore, was to investigate the etiology of this breeder mortality to map out possible critical periods during production in relation to possible risks of importance to the offspring. Dead birds from four flocks were subjected to postmortem and bacteriologic examination from onset of lay until slaughter (20-60 weeks). Causes of mortality were divided into noninfectious and infectious etiology. The infectious group could be subdivided into suppurative salpingitis/peritonitis caused by Escherichia coli and other infections (e.g., sepsis, endocarditis, and arthritis) mainly caused by Gram-positive cocci. Data analysis showed that 41% of the birds died from noninfectious causes, while 55% died from infectious causes, and 4% had no known cause of death. The prevalence of noninfectious mortality was highest in the youngest birds and lowest in the oldest birds. In contrast, the infectious mortality was lowest in the young birds and highest at the end of production. Within each age group, the prevalence of salpingitis/ peritonitis was 26% in young birds (20-29 weeks) and progressed throughout production to 41% in the oldest birds (≥50 weeks of age). Mortality due to other infections was low at onset of production (12%), peaking at 40-49 weeks of age (25%). Consequently, 40-49 weeks of age is identified as a critical period with regard to causes of mortality, possible vertical transmission of E. coli to the offspring, and increased risk of Gram-positive coccal infections.

Estudio longitudinal sobre las causas de mortalidad en reproductores pesados daneses. La producción de pollo de engorde depende en gran medida de la buena salud de las parvadas de reproductores. La llamada mortalidad normal en estas parvadas aún debe analizarse para reducir aún más la mortalidad de los reproductores y mejorar la calidad del pollo de engorde. El objetivo del presente estudio, por lo tanto, fue investigar la etiología de esta mortalidad en los reproductores para determinar posibles períodos críticos durante la producción con relación con los posibles riesgos que sean de importancia para la progenie. Las aves muertas de cuatro parvadas se sometieron a un examen post mortem y bacteriológico desde el inicio de la postura hasta la edad de sacrificio (20­60 semanas). Las causas de mortalidad se dividieron en etiología no infecciosa e infecciosa. El grupo de causas de origen infeccioso se subdividió en salpingitis/peritonitis supurativa (SP) causada por Escherichia coli y otras infecciones (por ejemplo, sepsis, endocarditis y artritis) causadas principalmente por cocos Gram positivos. El análisis de los datos mostró que el 41% de las aves murieron por causas no infecciosas, mientras que el 55% murió por causas infecciosas y el 4% por causas desconocidas. La prevalencia de mortalidad no infecciosa fue más alta en las aves más jóvenes y más baja en las aves con mayor edad. En contraste, la mortalidad infecciosa fue más baja en las aves jóvenes y más alta al final de la producción. Dentro de cada grupo de edad, la prevalencia de salpingitis/peritonitis supurativa fue del 26% en aves jóvenes (20­29 semanas) y aumentó a lo largo de la producción hasta el 41% en las aves con más edad (≥50 semanas de edad). La mortalidad debida a otras infecciones fue baja al inicio de la producción (12%), alcanzando un máximo de 40 a 49 semanas de edad (25%). En consecuencia, la edad entre 40 a 49 semanas se identifica como un período crítico con respecto a las causas de mortalidad, con la posible transmisión vertical de E. coli a la progenie y con el aumento del riesgo de infecciones por cocos Gram positivos.

Infectious potential of human derived uropathogenic Escherichia coli UTI89 in the reproductive tract of laying hens.

Avian pathogenic E. coli (APEC) and human uropathogenic E. coli (UPEC) harbour common virulence factors in spite of being associated with disease in different hosts. APEC strains have been shown to have zoonotic potential. In contrast, it is not known whether UPEC strains can cause infection in immunologically competent hens. The objective of the current study was to compare the ability of the well-characterized UPEC strain, UTI89, and the APEC strain, F149H1S2, to infect human and avian cells in culture and to cause salpingitis in an infection model in adult laying hens. In vitro characterization showed that the strains grew equally well in human urine, and both were able to infect human intestinal (Int407) and bladder (J82) epithelial cell lines, and they survived in avian macrophages (HD11) to the same extent. Groups of adult birds were inoculated with 108 bacteria directly into the oviduct using a surgical procedure. After an infection period of 48 h, bacterial load in the oviduct was determined by dilution series, and pathology was determined based on gross lesions and histological observations. Similar counts of UPEC UTI89 (ST95) and the APEC strain F149H1S2 (ST117) were obtained from tissues of infected birds, and salpingitis as evaluated by clinical score and histopathology was observed to a similar extent after infection with the two strains. Together, the results showed that UPEC UTI89 and APEC F149H1S2 have a similar potential for causing salpingitis in laying hens in the model used. No infection differences were observed between the UPEC UTI89 wild type and a mutant strain with knock-out of the well-known virulence gene, fimH, (UPEC UTI89ΔfimH), showing that the salpingitis model is not suitable for the detection of all UPEC virulence factors.

The SPI-19 encoded type-six secretion-systems (T6SS) of Salmonella enterica serovars Gallinarum and Dublin play different roles during infection.

Salmonella Pathogenicity Islands 19 (SPI19) encodes a type VI secretion system (T6SS). SPI19 is only present in few serovars of S. enterica, including the host-adapted serovar S. Dublin and the host-specific serovar S. Gallinarum. The role of the SPI19 encoded T6SS in virulence in these serovar is not fully understood. Here we show that during infection of mice, a SPI19/T6SS deleted strain of S. Dublin 2229 was less virulent than the wild type strain after oral challenge, but not after IP challenge. The mutant strain also competed significantly poorer than the wild type strain when co-cultured with strains of E. coli, suggesting that this T6SS plays a role in pathogenicity by killing competing bacteria in the intestine. No significant difference was found between wild type S. Gallinarum G9 and its ΔSPI19/T6SS mutant in infection, whether chicken were challenged orally or by the IP route, and the S. Gallinarum G9 ΔSPI19/T6SS strain competed equally well as the wild type strain against strains of E. coli. However, contrary to what was observed with S. Dublin, the wild type G9 strains was significantly more cytotoxic to monocyte derived primary macrophages from hens than the mutant, suggesting that SPI19/T6SS in S. Gallinarum mediates killing of eukaryotic cells. The lack of significant importance of SPI19/T6SS after oral and systemic challenge of chicken was confirmed by knocking out SPI19 in a second strain, J91. Together the results suggest that the T6SS encoded from SPI19 have different roles in the two serovars and that it is a virulence-factor after oral challenge of mice in S. Dublin, while we cannot confirm previous results that SPI19/T6SS influence virulence significantly in S. Gallinarum.

Haematology, blood biochemistry, parasites and pathology of common eider (Somateria mollisima) males during a mortality event in the Baltic.

A mortality event at the Christiansø colony in the Baltic proper killed 115 common eiders (Somateria mollissima) in mid-May 2016. To complement previous studies of incubating females, 39 males were necropsied and from a subsample of these a biochemical and haematological profile was obtained. The birds were emaciated and cachexic having a 50% reduction in body mass. Twenty-nine eiders were diagnosed with hydropericardium, 15 had hunger edema, three birds had enteritis and a single air sac infection. All birds were infested with intestinal Polymorphus minutus and 32 of these with the intestinal Trematoda spp. Microscopic parasitic investigations identified endoparasitic trematodes of the families Bucephhalidae, Echinostomidae, Notocotyluridae and Levinsiniella. White blood cell count showed slight heterophilia and lymphopenia while the albumin:globulin ratio (0.28) indicated stress, immune supression and inflammatory reactions supported by a high heterophil:lymphocyte index (13). Declined plasma concentration of glucose, fructosamine, amylase, albumin and protein likewise indicated long-term starvation prior to mortality indicating phase III starvation (catabolism of protein). The dramatic increase in aspartate transaminase, glutamate-dehydrogenase, lactate-dehydrogenase and bile acids indicate liver disorders while dehydration, renal, heart and bone disorders was reflected in the increased uric acid, urea, phosphor and potassium values. These findings show that male eiders undergo long-term starvation and multi organ failure similar to that of incubating females previously reported from the same colony. It increases our knowledge of the physiology of starving eiders and add to our understanding of the recurrent mortality events in the colony that seems to be linked to changes in food availability being an important factor together with a warmer climate in a declining Baltic eider population. We recommend future studies to focus on food composition, migration patterns and environmental changes including parasitic infections and global warming.

Longitudinal study of transmission of Escherichia coli from broiler breeders to broilers.

Escherichia coli is of major importance in industrial broiler production as the main cause of salpingitis and peritonitis in broiler breeders. Furthermore E. coli is the most common cause of first week mortality in broiler chickens. The aim of the present study was to investigate the transmission of E. coli, isolated from broiler breeders with salpingitis, to the progeny and the possibility of subsequent first week mortality. Four parent flocks were followed during the whole production period (20-60 weeks) by post mortem and bacteriological examination of randomly selected dead birds. Newly hatched chickens from each flock were swabbed in the cloaca on four occasions (parent age 30, 40, 50, 60 weeks) and E. coli was isolated. Causes of first week mortality were determined pathologically and bacteriologically. E. coli isolates from parents, newly hatched chickens and first week mortality were selected for Pulsed-Field-Gel-Electrophoresis (PFGE) and Multi-Locus-Sequence-Typing (MLST) to determine their clonal relationships. E. coli was the main cause of both salpingitis in parents and first week mortality in broilers, and E. coli dominated the bacterial flora of the cloaca of newly hatched chickens. PFGE of E. coli showed identical band patterns in isolates from the three different sources indicating a transmission of E. coli from parent birds to chickens. In conclusion, E. coli isolated from salpingitis in broiler parents were found to be transmitted to broilers in which some sequence types contributed to the first week mortality.