Erysipelothrix rhusiopathiae serotype 15 associated with recurring pig erysipelas outbreaks

Erysipelothrix rhusiopathiae is the causative agent of pig erysipelas and can be associated with sporadic cases or larger outbreaks of septicaemia with characteristic skin lesions or chronic polyarthritis. Within the genus Erysipelothrix, at least 6 species (Erysipelothrix rhusiopathiae, Erysipelothrix tonsillarum, Erysipelothrix species strain 1, Erysipelothrix species strain 2, Erysipelothrix species strain 3 and Erysipelothrix inopinata) and 28 serotypes (1a, 1b, 2-26 and N) have been recognised.1 E rhusiopathiae serotypes 1 and 2 are frequently isolated from clinically affected pigs, although other E rhusiopathiae serotypes have been sporadically associated with clinical disease. While there is no experimental evidence that Erysipelothrix species other than E rhusiopathiae cause disease in pigs, certain Erysipelothrix species strains have been isolated from clinical cases and from condemned carcases in abattoirs

Serotypes and Spa types of Erysipelothrix rhusiopathiae isolates from British pigs (1987 to 2015)

'Erysipelothrix' spp. cause a range of clinical signs in pigs and at least 28 different 'Erysipelothrix' spp. serotypes have been identified. In this study, 128 isolates of 'Erysipelothrix' spp. from pigs in Great Britain from 1987 to 2015 were characterised by serotyping and multiplex real time PCR assays targeting the surface protective antigen (Spa) and the main genotypes ('Erysipelothrix rhusiopathiae', 'Erysipelothrix tonsillarum' and 'Erysipelothrix' spp. strain 2). All 128 British isolates were characterised as 'E. rhusiopathiae' and were classified as serotypes 1a (n = 21), 1b (n = 17), 2 (n = 75), 5 (n = 2), 9 (n = 2), 10 (n = 2), 11 (n = 4) and 15 (n = 1), while four isolates were untypeable. All isolates were positive for the spa A gene. Serotypes 1a, 1b and 2 constituted 88.3% of the isolates; current serotype 2 based vaccines should protect against these isolates

PRRSV RNA detection in different matrices under typical storage conditions in the UK

In the UK, approximately 40 per cent of the pig breeding herds are outdoors. To monitor their porcine reproductive and respiratory syndrome virus (PRRSV) status, blood is collected commonly from piglets around weaning. Sample collection in British outdoor pigs often occurs during the early morning hours when the piglets tend to accumulate inside sheltered areas. For practical reasons, dry cotton swabs are occasionally used for blood collection and stored at room temperature until arrival in the laboratory. Detection of PRRSV RNA is a function of viral concentration, sample type and storage condition. To evaluate a possible impact of the sampling protocol on PRRSV1 detection, experimentally spiked blood samples using three dilutions of a representative PRRSV1 strain were prepared. In addition, blood samples from pigs naturally infected with PRRSV were obtained from a PRRSV-positive British herd. Spiked blood and blood from infected pigs were used to obtain sera, dry or wet (immersed in saline) polyester or cotton swabs and FTA cards. The different samples were stored for 24 hours, 48 hours or 7 days at 4°C or 20°C and tested by a real-time reverse transcriptase PRRSV PCR assay. Under the study conditions, the best matrix was serum (96.7 per cent), followed by wet swabs (78 per cent), dry swabs (61.3 per cent) and FTA cards (51 per cent). Polyester swabs (76 per cent) showed a better performance than cotton swabs (63.3 per cent). The reduction in sensitivity obtained for swabs and FTA cards was particularly high at low viral concentrations. The results indicate that wet polyester swabs should be used whenever possible

A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers

Melanin in pigmented organs like the skin is known to react with 3,3′-diaminobenzidine (DAB) to give a brown colour indistinguishable from the colour that DAB imparts to target antibodies bound to specific antigens. This can lead to false positives in chicken feathers during immunoperoxidase staining. Here, we present a simple, fast and practical method for bleaching chicken feathers which can be applied prior to immunohistochemistry staining without affecting specific antigen-antibody binding. To our knowledge, this is the first report of a melanin-bleaching technique prior to immunoperoxidase staining techniques of chicken feathers for detection of pathogens. Optimisations of the method include: • Removal of melanin from tissue sections using a short incubation with potassium permanganate followed by incubation with oxalic acid prior to immunostaining for improved specificity. • This technique did not affect the antigenicity of infectious laryngotracheitis virus antigen and did not cause damage or detachment of tissues from the slides

Genomic Sequence of a Megrivirus Strain Identified in Laying Hens in Brazil

A new strain of chicken megrivirus was identified in fecal samples of layer chickens in a commercial flock in Minas Gerais, Brazil. It is most closely related to the family Picornaviridae, genus Megrivirus, species Melegrivirus A, and has an overall nucleotide identity of up to 85.1% with other megrivirus strains

Microbial communities of poultry house dust, excreta and litter are partially representative of microbiota of chicken caecum and ileum

Traditional sampling methods for the study of poultry gut microbiota preclude longitudinal studies as they require euthanasia of birds for the collection of caecal and ileal contents. Some recent research has investigated alternative sampling methods to overcome this issue. The main goal of this study was to assess to what extent the microbial composition of non-invasive samples (excreta, litter and poultry dust) are representative of invasive samples (caecal and ileal contents). The microbiota of excreta, dust, litter, caecal and ileal contents (n = 110) was assessed using 16S ribosomal RNA gene amplicon sequencing. Of the operational taxonomic units (OTUs) detected in caecal contents, 99.7% were also detected in dust, 98.6% in litter and 100% in excreta. Of the OTUs detected in ileal contents, 99.8% were detected in dust, 99.3% in litter and 95.3% in excreta. Although the majority of the OTUs found in invasive samples were detected in non-invasive samples, the relative abundance of members of the microbial communities of these groups were different, as shown by beta diversity measures. Under the conditions of this study, correlation analysis showed that dust could be used as a proxy for ileal and caecal contents to detect the abundance of the phylum Firmicutes, and excreta as a proxy of caecal contents for the detection of Tenericutes. Similarly, litter could be used as a proxy for caecal contents to detect the abundance of Firmicutes and Tenericutes. However, none of the non-invasive samples could be used to infer the overall abundance of OTUs observed in invasive samples. In conclusion, non-invasive samples could be used to detect the presence and absence of the majority of the OTUs found in invasive samples, but could not accurately reflect the microbial community structure of invasive samples

Variability in practices for drinking water vaccination of meat chickens against infectious laryngotracheitis

Context: Drinking water vaccination of young meat chickens with Infectious Laryngotracheitis (ILT) vaccine is problematic. Vaccine failure and adverse vaccine reactions are frequently reported. Variations in the technique of applying ILT vaccines by this mass vaccination method need to be understood to contribute to improving the success of vaccination. Aims: This study aimed to examine variations in the techniques of application of Infectious Laryngotracheitis vaccines via drinking water for young meat chickens. Methods: Drinking water vaccination techniques were observed and recorded across 52 broiler flocks during ILT outbreaks in three geographic areas of Australia. Descriptive statistics for all variables were computed and variations between integrator company procedures were statistically compared. Key results: Despite rigorous standard operating procedures, wide variations were observed in time of water deprivation prior to vaccination (3–15 min), time drinking water was stabilised prior to addition of vaccine and the type of stabiliser product used, time to activate the flock following filling of the water lines with vaccine (10–127 min), time for the vaccine to be consumed (36–226 min) and the volume of drinking water per bird used to provide the vaccine (11–48 mL/bird). Conclusions: Variation in vaccination technique can affect the success of drinking water vaccination against ILT in young meat chickens. Implications: Understanding the importance of the variable factors in vaccine application method can improve the success of water vaccination against ILT