Fatal necrotic tracheitis by Aviadenovirus in captive Alagoas curassows (Pauxi mitu) extinct from the wild

Extinct from nature, captive young Alagoas curassows (Pauxi mitu) were found agonizing or dead with respiratory disease. Intranuclear inclusion bodies were found in the epithelia of the trachea, associated with marked necrotic tracheitis. An Aviadenovirus was isolated in chicken eggs and characterized genetically with 99% identity to the fowl Aviadenovirus A, as based on the hexon protein gene. This is the first report of respiratory disease caused by Aviadenovirus in any cracid species in Brazil, recommending for stricter biosecurity in the conservation premises.

RESEARCH HIGHLIGHTS

• Fatal tracheitis in curassows extinct from nature was associated with Aviadenovirus A.

• Seven-month-old Alagoas curassows (Aves: Cracidae) died with haemorrhagic tracheitis.

• Aviadenovirus A with 99% identity to fowl adenovirus 1 was detected in dead curassows.

• Fatal tracheitis by Aviadenovirus was described in Pauxi mitu (Aves: Cracidae).

     

Changes in gut bacterial communities in canaries infected by Macrorhabdus ornithogaster

Macrorhabdus ornithogaster is an opportunistic yeast that colonizes the gastric mucosa of many avian species. Until now, no studies have focused on the influence of a gastric infection on the balance of the intestinal microbiota of birds. In this study, 44 faecal samples from individual canaries, with and without M. ornithogaster infection, were analysed. The detection of the yeast was evaluated by 18S rRNA PCR. In order to evaluate the impact of the Macrorhabdus infection on the bacterial communities, culture-independent methods, by the use of amplicon-based sequencing as well as 16S rRNA-DGGE, were adopted. The different health status of animals affected the relative abundance of the main OTUs, with a greater diversification of the gut microbiota in healthy animals compared to the infected. In particular, Lactococcus, Pseudomonas, Acinetobacter, Lachnospiraceae, Propionibacterium and Weissella were found to be characteristic of uninfected animals (FDR?<?0.05), while Lactobacillus and Candidatus Arthromitus were characteristic of infected animals (FDR?<?0.05). Both these taxa have been reported as immunostimulatory, involved in immunological disorders. In infected animals the inferred metagenome assessed by PICRUST clearly showed a positive correlation between the presence of M. ornithogaster and KEGG genes related to ether lipid metabolism, already reported to be immunostimulatory by activation of macrophages and to play a pathophysiological role in several immunological disorders. Finally, our results show an interaction between infection of the digestive tract and intestinal microbiota of pet birds and provide insight into the changing of the complex enteric bacterial community.

• HIGHLIGHTS

• Macrorabdus ornithogaster is a gastric yeast that colonizes a wide range of birds.

• Differences were found between infected and healthy animals in gut microbiota.

• Candidatus Arthromitus was closely associated with infected birds.

• M. ornithogaster can affect intestinal microbiota composition of canaries.

     

Vaccination of chickens with the 34 kDa carboxy-terminus of Bpmp72 reduces colonization with Brachyspira pilosicoli following experimental infection

The anaerobic intestinal spirochaete Brachyspira pilosicoli colonizes the large intestine of a variety of species of mammals and birds, and may result in colitis, diarrhoea and reductions in growth rate. Naturally occurring infections in chickens are largely confined to adult laying and breeding birds. In this study, the 34 kD carboxy-terminus of the prominent outer membrane protein Bmp72 of B. pilosicoli was expressed as a histidine-tagged recombinant protein and used to immunize two groups (B and C) of 15 individually housed layer chickens. Vaccination was with either 100?μg (B) or 1?mg (C) protein emulsified with Freund’s incomplete adjuvant delivered into the pectoral muscles, followed three weeks later by 1?mg of protein in phosphate buffered saline delivered via crop tube. Two weeks later these and 15 non-vaccinated positive control birds (group A) housed in the same room were challenged via crop tube with B. pilosicoli avian strain CPS1. B. pilosicoli was detected in the faeces of all control birds and in 14 of the vaccinated birds in each vaccinated group at some point over the 30-day period following challenge. Colonization was delayed and the duration of excretion was significantly reduced (P?=?0.0001) in both groups of vaccinated birds compared to the non-vaccinated control birds. Fewer immunized birds had abnormal caecal contents at post mortem examination compared to non-vaccinated birds, but the difference was not statistically significant. This study indicates that recombinant Bmp72 C-terminus has potential to be developed for use as a vaccine component to provide protection against B. pilosicoli infections.

RESEARCH HIGHLIGHTS

• Laying chickens were immunized with recombinant Brachyspira pilosicoli membrane protein Bpmp72.

• Immunized birds had a highly significant reduction in the duration of colonization.

• Fewer immunized than control birds had abnormal caecal contents after infection.

• Bpmp72 showed potential for use as a novel vaccine component for B. pilosicoli.

     

Experimental model of oral transmissible AA amyloidosis in quails

ABSTRACT

In poultry and zoo birds, mass outbreaks of amyloid A (AA) amyloidosis are often reported, and horizontal transmission is considered as one of the causes. However, oral transmission of avian AA amyloidosis in nature has been unclear. In order to clarify the horizontal transmission of avian AA amyloidosis, basic research using an appropriate oral transmission model is necessary. In this study, we developed an oral transmission model of AA amyloidosis using quails, and assessed the oral transmission efficiency of AA amyloidosis in quails and mice. Young quails, adult quails, and young mice received inflammatory stimulation with lipopolysaccharide; simultaneously, homogeneous amyloid fibrils were orally or intravenously administered. By histological examination, induction of amyloidosis by oral or intravenous administration of amyloid was confirmed in all species. Furthermore, both quail and murine AA amyloidosis were orally transmitted in a dose-dependent manner. These results support the possibility of horizontal transmission of avian AA amyloidosis in nature. This model will be able to contribute to the elucidation of spontaneous horizontal transmission of avian AA amyloidosis in the future.

RESEARCH HIGHLIGHTS

• Quail AA amyloidosis was orally transmitted in a dose-dependent manner.

• Oral transmission was less efficient than intravenous transmission.

• In-cage horizontal transmission did not occur during 4-week cohabitation.

• Amyloid deposition in tissues of quail was grossly visible.

     

MinION sequencing to genotype US strains of infectious laryngotracheitis virus

Over the last decade the US broiler industry has fought long-lasting outbreaks of infectious laryngotracheitis (ILTV). Previously, nine genotypes (I-IX) of ILTVs have been recognized using the polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) method with three viral alleles (gB, gM and UL47/gG). In this study, the genotyping system was simplified to six genotypes by amplicon sequencing and examining discriminating single nucleotide polymorphisms (SNPs) within these open reading frames. Using phylogenomic analysis of 27 full genomes of ILTV, a single allele (ORF A/ORF B) was identified containing SNPs that could differentiate ILTVs into genotypes congruent with the phylogenetic partitioning. The allelic variations allowed for the cataloging of the 27 strains into 5 genotypes: vaccinal TCO, vaccinal CEO, virulent CEO-like, virulent US and virulent US backyard flocks from 1980 to 1990, correlating with the PCR-RFLP genotypes I/ II/ III (TCO), IV (CEO), V (virulent CEO-like), VI (virulent US) and VII/VIII/IX (virulent US backyard flock isolates). With the unique capabilities of third generation sequencing, we investigated the application of Oxford Nanopore MinION technology for rapid sequencing of the amplicons generated in the single-allele assay. This technology was an improvement over Sanger-based sequencing of the single allele amplicons due to a booster amplification step in the MinION sequencing protocol. Overall, there was a 90% correlation between the genotyping results of the single-allele assay and the multi-allele assay. Surveillance of emerging ILTV strains could greatly benefit from real-time amplicon sequencing using the single-allele assay and MinION sequencing.

RESEARCH HIGHLIGHTS

• A multi-allelic assay identified nine ILTV genotypes circulating in the US

• Single-allele genotyping is congruent with whole genome phylogenetic partitioning

• US ILTV strains can be grouped into five genotypes using the single-allele assay

• The single-allele assay can be done using MinION sequencing of barcoded amplicons

     

New PA/1220/98-like variant of infectious bronchitis virus in Poland

ABSTRACT

The aim of the present study was to report the first detection of a new infectious bronchitis virus (IBV) variant in Polish commercial flocks which is completely different to any previously known in this region. In 2018, samples from Ross 308 breeding hens aged 35 weeks were delivered for IBV diagnosis. IBV presence was detected, but all attempts to amplify the S gene fragment were negative. The field material was analysed using the Illumina MiSeq platform and a 1073-nt fragment of the S1 coding region was obtained. The gCoV/ck/Poland/516/2018 strain shared only 52.7–58.1% nucleotide identity to any known genotype of IBV and shared the highest identity of 81.4% to the unique North American PA/1220/98 variant. Based on the obtained sequence, a specific molecular test was constructed and used for screening of chicken samples from 35 field cases delivered to our laboratory between 2018 and 2019 for IBV diagnosis. Application of this test enabled detection of another three chicken flocks as positive for this new strain. All positives were identified in commercial layers with egg production problems. To date, the virus has not been detected in broiler chickens. Taking into account the proposed criteria for the definition of a new IBV genotype or lineage, it seems that the detected viruses in Poland, together with the unique North American PA/1220/98 variant, may be classified as separate lineages/genotype in the new IBV classification.

RESEARCH HIGHLIGHTS

• The new IBV variant is distantly related to other known GI–GVII IBV genotypes/lineages.

• It affects long-lived birds causing egg production problems.

• The detected IBV and the unique North American PA/1220/98 variant are candidates for separate lineages in the new GVIII genotype.

     

Dermanysuss gallinae attacks humans. Mind the gap!

ABSTRACT

Dermanyssus gallinae is a haematophagous ectoparasite primarily known as a pest of domestic and wild birds. It occasionally feeds on a range of mammals, and, more importantly, is of growing concern in human medicine. This review highlights mite attacks on people working with poultry, and updates the increasing incidence of dermanyssosis in urban environments in Europe. Although several cases of dermanyssosis have been documented, there are a number of reasons why diagnosis of D. gallinae infestations in humans is likely to be underestimated. Firstly, medical specialists are not well aware of D. gallinae infestations in humans. There is also a lack of collaboration with specialists from other disciplines. The problem is compounded by misdiagnoses and by the lack of diagnostic tools. We review the literature on human dermanyssosis cases in Europe, and also provide information on the epidemiology, clinical, histo-pathological and immunological aspects of dermanyssosis. We stress the need for improved recognition of this challenging infestation in humans, and provide straightforward recommendations for health practitioners, starting with collection of the correct anamnestic information and including appropriate management methods for case recognition and resolution. Finally, we indicate the most urgent areas to be addressed by future research.

RESEARCH HIGHLIGHTS

• Dermanyssus gallinae is of growing concern in human medicine.

• Most physicians are not well aware of dermanyssosis in humans.

• Bio-epidemiological and clinical aspects of this ectoparasitosis are highlighted.

• Practical key actions for diagnosis and correct management of infestation in humans are provided.

     

Pathology of clade 2.3.2.1 avian influenza virus (H5N1) infection in quails and ducks in Bangladesh

We performed pathological and molecular virological investigation of three outbreaks of highly pathogenic avian influenza (HPAI) in a quail farm and two duck farms of Mymensingh and Netrokona districts of Bangladesh in 2011. HPAI viruses of subtype H5N1 were detected from all three outbreaks and phylogenetic analysis of HA gene sequence placed the viruses into clade 2.3.2.1. The outbreak in the quail farm was characterized by acute death with 100% mortality within two days. Marked haemorrhages and congestion with necrotic and inflammatory lesions in the respiratory tract, liver, pancreas and kidneys were the major gross and histopathological lesions. In the case of ducks, nervous signs were the remarkable clinical manifestations and the mortality was around 10%. No significant gross lesions were observed at necropsy. Non-purulent encephalitis with gliosis and neuronal degeneration was observed on histopathological examination. By immunohistochemistry, viral antigen could be detected in different organs of both quails and ducks. This study records varying clinical and pathological manifestations of HPAI in ducks and quails following natural infection with the same strain of the virus.

RESEARCH HIGHLIGHTS

• HPAIV of clade 2.3.2.1 was detected from clinical outbreaks in quails and ducks

• Sudden death with severe haemorrhages in various organs was found in quails

• Pronounced nervous signs with non-purulent encephalitis were observed in ducks

• Viral antigen could be localized in different organs by immunohistochemistry

     

How can Dermanyssus gallinae (De Geer 1778) (Acari: Anactinotrichida: Dermanyssidae) walk upwards on slippery surfaces?

ABSTRACT

Scanning electron microscopy observations of the distal leg region of the poultry red mite Dermanyssus gallinae (De Geer 1778) identified the presence of a compound ambulacrum, the part of the leg that contacts the substratum when the mite walks. The ambulacrum is comprised of a praetarsus (the ambulacrum stalk), a pulvillus and two claws. The pulvillus is a weakly sclerotized structure that can be partly expanded or retracted in the praetarsus. When expanded, the pulvillus shows a cushion-like shape which can, as a result of its soft surface, function as a sucker, thus allowing D. gallinae to adhere to a smooth surface. When traversing an irregular surface, or clinging to a soft surface, the mite retracts the pulvillus and uses only its strongly sclerotized movable claws. These observed morphological adaptations explain the ability of D. gallinae to climb upwards on a slippery surface, resist an air flux, walk on smooth and rigid feathers of its avian hosts, and cling to the bird’s or human's soft skin. This knowledge is important to better understand the attachment mechanism of this species to its host and to the substratum on which it moves, and also to provide insight into the circumstances under which it is able to move.

RESEARCH HIGHLIGHTS

• The ambulacrum is the distal part of the leg touching the substratum

• D. gallinae’s ambulacrum consists of a praetarsus, a pulvillus and two claws

• The weakly sclerotized pulvillus can be part expanded/retracted in the praetarsus

• The expanded pulvillus functions as a sucker to adhere to smooth surfaces

• The claws are used to walk on an irregular surface or cling to a soft surface

     

Retrospective study on transmissible viral proventriculitis and chicken proventricular necrosis virus (CPNV) in the UK

ABSTRACT

Chicken proventricular necrosis virus (CPNV) is a recently described birnavirus, which has been proposed to be the cause of transmissible viral proventriculitis (TVP). The understanding of the epidemiology of both the virus and the disease is very limited. A retrospective investigation on TVP and CPNV in broiler chicken submissions from the UK from between 1994 and 2015 was performed with the aims of assessing the longitudinal temporal evolution of TVP and CPNV, and to review the histological proventricular lesions in the studied chickens. Ninety-nine of the 135 included submissions (73.3%) fulfilled the TVP-diagnostic criteria, while the remaining 36 submissions (26.7%) displayed only lymphocytic proventriculitis (LP). The first detection of CPNV by PCR dated from 2009. Results showed a rise in the number of both TVP and positive CPNV RT–PCR submissions from 2009 with a peak in 2013, suggesting that they may be an emerging or re-emerging disease and pathogen, respectively. Twenty-two out of the 99 submissions displaying TVP lesions (22%) and four out of the 36 (11%) submissions with LP gave positive CPNV RT–PCR results, further supporting the association between CPNV and TVP and confirming that CPNV is present in a low proportion of proventriculi that do not fulfil the TVP-diagnostic criteria. In addition, intranuclear inclusion bodies were observed in 22 of the submissions with TVP. The vast majority of these cases (21 of 22, 96%) gave negative CPNV RT–PCR results, raising the question of whether a virus other than CPNV is responsible for some of these TVP-affected cases.

RESEARCH HIGHLIGHTS

• TVP and CPNV have been present in British broilers since at least 1994 and 2009, respectively.

• TVP and CPNV seem to be an emerging and re-emerging disease and pathogen, respectively.

• CPNV was detected in proventriculi with both TVP and LP-lesions.

• Viruses other than CPNV may be responsible for some TVP-affected cases.

     

Antigenicity,pathogenicity and immunosuppressive effect caused by a South American isolate of infectious bursal disease virus belonging to the “distinct” genetic lineage

Infectious bursal disease virus (IBDV) is the causative agent of a highly contagious immunosuppressive disease affecting young chickens. The recently described “distinct IBDV” (dIBDV) genetic lineage encompasses a group of worldwide distributed strains that share conserved genetic characteristics in both genome segments making them unique within IBDV strains. Phenotypic characterization of these strains is scarce and limited to Asiatic and European strains collected more than 15 years ago. The present study aimed to assess the complete and comprehensive phenotypic characterization of a recently collected South American dIBDV strain (1/chicken/URY/1302/16). Genetic analyses of both partial genome segments confirmed that this strain belongs to the dIBDV genetic lineage and that it is not a reassortant. Antigenic analysis with monoclonal antibodies indicated that this strain has a particular antigenic profile, similar to that obtained in a dIBDV strain from Europe (80/GA), which differs from those previously found in the traditional classic, variant and very virulent strains. Chickens infected with the South American dIBDV strain showed subclinical infections but had a marked bursal atrophy. Further analysis using Newcastle disease virus-immunized chickens, previously infected with the South American and European dIBDV strains, demonstrated their severe immunosuppressive effect. These results indicate that dIBDV strains currently circulating in South America can severely impair the immune system of chickens, consequently affecting the local poultry industry. Our study provides new insights into the characteristics and variability of this global genetic lineage and is valuable to determine whether specific control measures are required for the dIBDV lineage.

Research Highlights

• A South American strain of the dIBDV lineage was phenotypically characterized.

• The strain produced subclinical infections with a marked bursal atrophy.

• Infected chickens were severely immunosuppressed.

• The dIBDV strains are antigenically divergent from other IBDV lineages.

     

Synergistic effect of a combination of nicarbazin and monensin against coccidiosis in the chicken caused by Eimeria spp.

ABSTRACT

A clinical study was made into the abilities of nicarbazin and monensin and a nicarbazin?+?monensin combination to control Eimeria acervulina, E. maxima, and E. tenella in chickens. When included in the feed, at concentrations of 40?ppm nicarbazin or 40?ppm monensin, these products showed partial efficacy evaluated by daily weight gain (DWG) but no activity judged by daily feed intake (DFI) or feed conversion ratio (FCR). By contrast, the combination of 40?ppm nicarbazin?+?40?ppm monensin provided complete control of infection judged by greater DWG and DFI, and lower FCR. Monensin at a concentration of 40?ppm was ineffective in preventing lesions caused by all three species. Nicarbazin at a concentration of 40?ppm was unable to suppress lesions of E. acervulina and E. maxima but was able to suppress lesions caused by E. tenella. Nicarbazin 40?ppm?+?monensin 40?ppm suppressed lesions of all three species.

RESEARCH HIGHLIGHTS

• Nicarbazin or monensin at 40 ppm gave only partial control of Eimeria spp.

• A combination of 40 ppm nicarbazin + 40 ppm monensin controlled DWG, DFI and FCR.

• Nicarbazin or monensin at 40 ppm did not suppress all Eimeria spp. lesions.

• Nicarbazin 40 ppm + monensin 40 ppm suppressed lesions of all three species.

     

Genotyping of Riemerella anatipestifer by ERIC-PCR and correlation with serotypes

Riemerella anatipestifer (RA) is a widely distributed bacterial pathogen of birds responsible for remarkable losses to poultry production, especially among waterfowl. We characterized the genomic diversity of 166 field isolates of RA, collected from geese and ducks, using enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR). The field strains and five reference strains showed 17 distinct patterns consisting of five to 12 bands ranging from approximately 150–1800bp. The majority of the strains belonged to two closely related ERIC-PCR types (A and B), while the other types contained only a few isolates each. There was no association between ERIC-PCR type and host species, place, or year of isolation; however the ERIC-PCR pattern was correlated with serotype for most isolates. The majority of serotype 1 strains (101/107) belonged to ERIC-PCR type A while the remaining six strains represented five different ERIC-PCR types (D, G, L, M, and O). Serotypes 1,7 and 7 corresponded to ERIC-PCR types B and C, respectively. Serotypes 2, 4, and 10 could be subdivided by ERIC-PCR revealing two to four patterns within each serotype. These results indicate that ERIC-PCR may be a suitable technique for the molecular identification of RA serotypes, and the detection of subtypes within certain serotypes may aid further epidemiological investigations.

RESEARCH HIGHLIGHTS

• ERIC-PCR analysis of field R. anatipestifer strains revealed 17 distinct patterns

• Most strains belonged to two closely related ERIC-PCR types

• Serotype 1 was the most prevalent serotype representing 64.5% of the strains

• ERIC-PCR may be suitable for molecular identification of R. anatipestifer serotypes

     

Biological control as an alternative to synthetic fungicides for the management of grey and blue mould diseases of table grapes: a review

Abstract

Grey and blue mould diseases are among the most important diseases of grapes worldwide. They are causing extensive decay in postharvest grapes. Chemical fungicides remain the primary treatment for managing these diseases. However, consumer’s interest in organic produce, restrictions on chemical use by some countries, and concerns about human and environmental safety have driven research to identify safe and effective alternatives. Among several alternative approaches, the use of biological control agents (BCAs) is getting more acceptance and has been comprehensively studied. This review summarizes the use of BCAs as a postharvest treatment to control the postharvest grey and blue mould of grapes. The review also emphasizes the economic importance of these two major postharvest diseases of grapes. In addition, other non-chemical postharvest treatments, the advantage of an integrated approach and finally problems, challenges, and future trends of the BCAs are described. Several yeasts that have a promising result to control grey and blue mould disease of grapes are thoroughly reviewed. The current market share of BCAs and their future directions on commercialization are also suggested in this review.

• Biocontrols can be a potential control method for postharvest diseases of fruits and vegetables in place of chemical fungicides.

• Antagonistic yeasts have high ability to suppress fungal growth.

• Integrated approach utilizes a combination approaches, which often results in a synergistic effect to control the diseases.

• USA and Europe are currently the largest markets for biocontrol products.

     

Pathogenicity and genomic characterization of a novel avian orthoreovius variant isolated from a vaccinated broiler flock in China

Avian orthoreovirus (ARV) infections of broiler flocks cause arthritis/tenosynovitis syndrome and significant economic losses. ARV variants were detected in the USA and Canada. Viral arthritis/tenosynovitis syndrome has occurred frequently in China in recent years. In this study, a variant ARV strain associated with viral arthritis/tenosynovitis syndrome was isolated from broilers and designated as LY383. Genomic sequence and phylogenetic analysis of the σC nucleic acid and amino acid sequences revealed that the isolate was closely related to ARV field strains Reo/PA/Layer/01224B/14, Reo/PA/Broiler/1551/13, GA/14602/2014, GA/13569/2013 and GA/13542/2013, in cluster V, but distinct from most Chinese field strains or commercial vaccine strains. Experimental challenge showed that the isolate could cause arthritis/tenosynovitis syndrome in broilers, which possessed a high level of maternal antibodies induced by commercial ARV vaccines (S1133, 1733 and T98). Furthermore, viral nucleic acid could be detected in cloacal swabs of all challenged birds throughout the entire test from 5?dpi onward. These results suggest that a novel ARV genotype emerges and might become prevalent in broiler flocks in China.

RESEARCH HIGHLIGHTS

• A variant avian orthoreovirus was isolated from a vaccinated broiler flock in North China.

• The ARV field strain was distinct from previous China-origin ARV isolates and vaccine strains.

• The current commercial ARV vaccine could not provide effective protection of broilers against the field isolate infection.

• These findings indicated that variant ARV field strains might become frequent in broiler flocks in China and effective measures should be conducted to prevent and control the disease.

     

Detection of Histomonas meleagridis DNA in dust samples obtained from apparently healthy meat turkey flocks without effect on performance

Environmental dust samples obtained from 65 turkey flocks in France, of which six suffered from histomonosis whereas the rest remained apparent healthy until the end of production, were tested for the presence of Histomonas meleagridis DNA by recently developed real-time PCR based on the 18S rRNA locus. In order to determine the genotype of detected histomonads, positive samples were further subjected to conventional 18S PCR and sequencing. Additionally, production data of all tested flocks, such as average daily gain, feed conversion ratio and production index, were statistically evaluated and compared to see the effect of positive dust samples in apparently healthy flocks. Histomonad DNA was detected in the dust obtained from all six clinically affected and, surprisingly, in nine apparent healthy flocks. Sequencing of the 18S rRNA gene resulted in only one DNA sample homologous to H. meleagridis whereas 11 others revealed the presence of several other flagellates. Average daily gain and production index were negatively affected in flocks with clinical histomonosis, resulting in significant difference in comparison with the data obtained from clinically healthy flocks independent of the presence of histomonad DNA in the dust. Overall, there was no significant difference following statistical analysis of production parameters between the two last mentioned groups of tested flocks. Altogether, this is the first investigation demonstrating the presence of H. meleagridis DNA in environmental dust samples obtained from clinically unaffected turkey flocks. However, this finding could not be correlated with impact on production based on analysis and comparison of selected production data.

RESEARCH HIGHLIGHTS

• Environmental dust obtained from clinically healthy turkey flocks, in addition to dust from flocks affected by histomonosis, was found positive for the presence of Histomonas meleagridis DNA.

• Histomonas-positive dust samples in clinically unaffected flocks did not have a negative effect on production parameters.

• The results demonstrate a wider spread of H. meleagridis DNA in flocks of commercial meat turkeys than previously thought.

     

Pigeon circoviruses from feral pigeons in Australia demonstrate extensive recombination and genetic admixture with other circoviruses

ABSTRACT

Like other avian circovirus species, Pigeon circovirus (PiCV) is known to be genetically diverse with a relatively small circular single-stranded DNA genome of 2?kb that encodes for a capsid protein (Cap) and a replication initiator protein (Rep). Recent paleoviral evidence hints towards a probable Gondwanan origin of avian circoviruses, paralleling the evolution and dispersal of their hosts. Limited availability of PiCV genome sequence data in Australia has hindered phylogeographic studies in this species, so we screened clinically normal rock doves (Columba livia) in regional New South Wales, and demonstrated a high prevalence (76%) of PiCV infection by PCR. We also recovered 12 complete novel PiCV genomes and phylogenetic analyses revealed that PiCV circulating in Australian feral pigeons formed two strongly supported monophyletic clades. One clade resided with PiCV genomes from Poland, Australia, United Kingdom, Belgium, China, and Japan, and another basal clade was more closely related to PiCV genomes from Poland. A novel more distantly-related PiCV rep gene formed a solitary clade with weak posterior support. So we further analysed all selected partial rep gene sequences to demonstrate a likely naturally occurring spillover infection from a passerine circovirus candidate. The findings suggest that there is a high degree of genetic variation within PiCV in Columbiformes with potential greater admixture between avian circoviruses within Australia than previously known.

RESEARCH HIGHLIGHTS

• Confirmed high prevalence rate of PiCV circulating in Australian wild pigeons.

• Highlighted extensive recombination events within Australian PiCV.

• Demonstrated a likely naturally occurring spillover infection from a passerine circovirus candidate.

     

Anatomopathological changes,quantification and distribution of Libyostrongylus spp. in regions of the proventriculus and ventriculus of naturally- and experimentally-infected ostriches

Nematodes of the genus Libyostrongylus parasitize ostriches, causing high mortality rates. These nematodes are found in the proventriculus and ventriculus of ostriches, but little is known about their distribution and the possible anatomopathological changes they cause in the various regions of these organs. This paper describes the distribution and quantification of Libyostrongylus and pathological changes found in regions of the proventriculus and ventriculus of ostriches with high and low levels of both natural and experimental infection. Ostriches were necropsied and tissue samples from the distinct regions of both organs were analysed based on nematode counts and histopathology after staining with haematoxylin and eosin, Masson’s trichrome or Alcian blue/PAS. The cranial and glandular regions of the proventriculus were the most parasitized. The ventriculus contained more nematodes in the caudal region. No macro- or microscopic pathological changes were observed in either of these organs of experimentally-infected birds. However, naturally-infected birds with high levels of infection presented proventriculus with macroscopic lesions and heterophilic infiltrates surrounding nematodes. In the glandular region of this organ, nematodes were located in the adenomeres of the secretory ducts, causing altered architecture and erosions and ulcerative lesions with damaged epithelium. Nematode eggs were found in the koilin layer of the middle and caudal regions of the ventriculus only of these birds. The pH of the regions assessed by Alcian blue/PAS staining changed from acidic in the proventriculus to more alkaline in the caudal region of the ventriculus. These data add knowledge to the biology of Libyostrongylus.

RESEARCH HIGHLIGHTS

• The most parasitized areas were the cranial and glandular regions of the proventriculus.

• Naturally-infected birds with high levels of infection presented macro lesions in the proventriculus and damaged epithelium.

• Nematode eggs were found in the ventriculus.

• The proventriculus had an acidic pH, which turned alkaline towards the ventriculus.

     

Physiological,biochemical and histopathological changes associated with heatstroke in the galah (Eolophus roseicapilla) and rock dove (Columba livia)

The pathophysiology of heat illnesses in birds has not been well characterized. In this study, we describe the changes in heart rate, respiratory rate, blood biochemistry and histopathological findings in galahs and rock doves after heat exposure under standardized conditions designed to induce heatstroke. Birds in the heat-exposed group were exposed to environmental heat stress and compared to control birds. Both groups of birds were under general anaesthesia throughout the experiment and serial blood collections were performed for biochemical analyses, while organs were collected at the end of the experiment for histopathology. No electromyography traces consistent with the onset of heat cramps were observed in any of the birds. Biochemical changes suggestive of skeletal muscle and hepatocellular injury, including hyperkalaemia and increased serum muscle and hepatic enzyme activities, were often observed in heat-exposed galahs and rock doves at the onset of heatstroke. Microscopic analyses did not reveal any significant cardiac changes, although some lungs had signs of acute congestion. Some heat-exposed rock doves had microscopic changes indicative of necrosis in the pectoral muscle. There were significant hepatic changes in some heat-exposed galahs, but not in rock doves. This suggests that there may be species differences amongst birds in the organs most affected by heatstroke. The observed species differences in the physiological, biochemical and histopathological changes indicate that bird species should be studied separately for clinical syndromes such as heatstroke.

RESEARCH HIGHLIGHTS

• Biochemical changes suggestive of skeletal muscle and hepatocellular injury in heat-exposed galahs and rock doves at the onset of heatstroke

• No electromyography traces consistent with the onset of heat cramps were observed

• Some heat-exposed rock doves had indications of necrosis in the pectoral muscle

• There were significant hepatic changes in some heat-exposed galahs

     

Metaproteomics characterization of the alphaproteobacteria microbiome in different developmental and feeding stages of the poultry red mite Dermanyssus gallinae (De Geer, 1778)

ABSTRACT

The poultry red mite (PRM), Dermanyssus gallinae (De Geer, 1778), is a worldwide distributed ectoparasite and considered a major pest affecting the laying hen industry in Europe. Based on available information in other ectoparasites, the mite microbiome might participate in several biological processes and the acquisition, maintenance and transmission of pathogens. However, little is known about the role of PRM as a mechanical carrier or a biological vector in the transmission of pathogenic bacteria. Herein, we used a metaproteomics approach to characterize the alphaproteobacteria in the microbiota of PRM, and variations in its profile with ectoparasite development (nymphs vs. adults) and feeding (unfed vs. fed). The results showed that the bacterial community associated with D. gallinae was mainly composed of environmental and commensal bacteria. Putative symbiotic bacteria of the genera Wolbachia, C. Tokpelaia and Sphingomonas were identified, together with potential pathogenic bacteria of the genera Inquilinus, Neorickettsia and Roseomonas. Significant differences in the composition of alphaproteobacterial microbiota were associated with mite development and feeding, suggesting that bacteria have functional implications in metabolic pathways associated with blood feeding. These results support the use of metaproteomics for the characterization of alphaproteobacteria associated with the D. gallinae microbiota that could provide relevant information for the understanding of mite-host interactions and the development of potential control interventions.

Research highlights

• Metaproteomics is a valid approach for microbiome characterization in ectoparasites.

• Alphaproteobacteria putative bacterial symbionts were identified in D. gallinae.

• Mite development and feeding were related to variations in bacterial community.

• Potentially pathogenic bacteria were identified in mite microbiota.