Prevalence of <Emphasis Type="Italic">Chlamydophila psittaci</Emphasis> in wild birds—potential risk for domestic poultry,pet birds,and public health?

To determine the prevalence of Chlamydophila psittaci in wild birds, cloacal swabs from 527 songbirds, 442 waterfowl, 84 feral pigeons, and 38 cormorants were examined by Chlamydiaceae-specific real-time polymerase chain reaction (PCR) and ArrayTube microarray assays for chlamydial species determination and genotyping of C. psittaci. Inconclusive cases were further characterized by conventional PCR methods targeting the chlamydial outer membrane protein A, 16S, 23S, and intergenic spacer genes followed by sequencing of the PCR product. Swabs of 19 water birds (tufted ducks and pochards), 12 pigeons, and one songbird were tested positive by the Chlamydiaceae-specific real-time PCR. While C. psittaci genotypes B (n = 5) and E (n = 1) were identified in feral pigeons (n = 9), the genotype could not be identified in the remaining three cases. Sequence data of Chlamydiaceae-positive cases (n = 23; 19 waterfowl, three pigeons, one songbird) indicated the presence of nonclassified chlamydial agents (n = 20) and C. psittaci (n = 3) by 16S rRNA PCR and sequencing. In conclusion, C. psittaci was not detected in waterfowl and songbirds, but C. psittaci proved prevalent in urban feral pigeons, where it poses a significant risk for humans.     

Populations and trends of Canadian Arctic seabirds

Canada’s eastern Arctic (Nunavut and Arctic Quebec—Nunavik, N of 60°) supports large numbers of seabirds in summer. Seabird breeding habitat in this region includes steep, rocky coasts and low-lying coasts backed by lowland sedge-meadow tundra. The former areas support colonial cliff- and scree-nesting seabirds, such as murres and fulmars; the latter inland or coastal seabirds, such as terns, gulls and jaegers. The region supports some 4 million breeding seabirds, of which the most numerous are thick-billed murres (Uria lomvia; 75%), black guillemots (Cepphus grylle; 9%), northern fulmars (Fulmarus glacialis; 8%) and black-legged kittiwakes (Rissa tridactyla; 6%). The majority of Arctic seabirds breed in a small number of very large colonies (>10,000 birds), but there are also substantial numbers of non-colonial or small-colony breeding populations that are scattered more widely (e.g. terns, guillemots). Population trends among Canadian Arctic seabirds over the past few decades have been variable, with no strongly negative trends except for the rare ivory gull (Pagophila eburnea): this contrasts with nearby Greenland, where several species have shown steep declines. Although current seabird trends raise only small cause for concern, climate amelioration may enable increased development activities in the north, potentially posing threats to some seabirds on their breeding grounds.     

The Chlamydia psittaci genome: a comparative analysis of intracellular pathogens

Background

Chlamydiaceae are a family of obligate intracellular pathogens causing a wide range of diseases in animals and humans, and facing unique evolutionary constraints not encountered by free-living prokaryotes. To investigate genomic aspects of infection, virulence and host preference we have sequenced Chlamydia psittaci, the pathogenic agent of ornithosis.

Results

A comparison of the genome of the avian Chlamydia psittaci isolate 6BC with the genomes of other chlamydial species, C. trachomatis, C. muridarum, C. pneumoniae, C. abortus, C. felis and C. caviae, revealed a high level of sequence conservation and synteny across taxa, with the major exception of the human pathogen C. trachomatis. Important differences manifest in the polymorphic membrane protein family specific for the Chlamydiae and in the highly variable chlamydial plasticity zone. We identified a number of psittaci-specific polymorphic membrane proteins of the G family that may be related to differences in host-range and/or virulence as compared to closely related Chlamydiaceae. We calculated non-synonymous to synonymous substitution rate ratios for pairs of orthologous genes to identify putative targets of adaptive evolution and predicted type III secreted effector proteins.

Conclusions

This study is the first detailed analysis of the Chlamydia psittaci genome sequence. It provides insights in the genome architecture of C. psittaci and proposes a number of novel candidate genes mostly of yet unknown function that may be important for pathogen-host interactions.     

Chlamydia buteonis,a new Chlamydia species isolated from a red-shouldered hawk

Chlamydiaceae are obligate intracellular bacterial pathogens for humans and animals. A recent study highlighted that a Chlamydiaceae intermediary between C. psittaci and C. abortus can infect hawks. Here, an isolate was obtained upon passage of cloacal and conjunctival sac material collected from a female hatch-year red-shouldered hawk (Buteo lineatus) in cultured cells. The diseased bird, one of 12 birds housed in a rehabilitation center, developed conjunctivitis and later died. Swabs from both sites tested positive for Chlamydia using the QuickVue Chlamydia test. The isolate, named RSHA, tested negative in qPCR assays specific for C. psittaci and C. abortus, respectively. Analysis of the 16S rRNA, 23S rRNA and whole genome sequences as well as MLST, ANIb and TETRA values reveal that C. psittaci and C. abortus are the closest relatives of RSHA. However, the overall results strongly suggest a phylogenetic intermediate position between these two species. Therefore, we propose the introduction of a new species designated Chlamydia buteonis with RSHA^T as the type strain.     

The influence of predation by herring gullsLarus argentatus and oystercatchersHaematopus ostralegus on a newly established musselMytilus edulis bed in autumn and winter

Predation by herring gullsLarus argentatus and oystercatchersHaematopus ostralegus was evaluated on a newly established musselMytilus edulis bed on tidal flats of the German Wadden Sea. The mussel bed covered an area of 2 ha and showed a decrease in biomass of 40% in the most densely covered parts from August to January. Synchronously, the extent of the mussel bed was reduced, resulting in a decrease of average biomass of 98% over the whole mussel bed. From the beginning of August 1994 to mid January 1995, the average size of mussels increased from 10.7 to 20.3 mm. The P/B-ratio was 0.68 in August and 0.18 between September and November. Herring gulls and oystercatchers were the most important mussel predators. On average, 266 herring gulls and 63 oystercatchers were present on the mussel bed during one low tide; 34% of the herring gulls and 78% of the oystercatchers were observed to be feeding. Herring gulls fed at a rate of 4.2 mussels per minute and oystercatchers at a rate of 1.3 mussels per minute. While herring gulls took the most common mussel sizes (mean: 20 mm), oystercatchers searched for the largest mussels available (mean: 25 mm). Herring gulls consumed 13 mussels/m² (0.3g AFDW) during one day and oystercatchers 1.7 mussels/m² (0.1 g AFDW). Predation by birds was compensated by 33% of the production. The proportion removed by bird predation amounted to 10% of abundance and to 16% of biomass (including production). Oystercatchers were responsible for 1% of the reduction in abundance and for 3% of biomass. Removal was highest in the most common size classes of mussels, mainly caused by herring gulls. However, the highest proportion of mussels was eaten in the largest size classes, mainly by oystercatchers. *** DIRECT SUPPORT *** A03B6035 00004     

Study of protein metabolism of herring gulls (<Emphasis Type="Italic">Larus argentatus</Emphasis> Pontop.) infected by trematode <Emphasis Type="Italic">Himasthla larina</Emphasis> (Trematoda: Echinostomatidae)

The values and dynamics of some indices of protein metabolism were studied in herring gulls Larus argentatus infected with trematode Himasthla larina in natural populations and in experiment. These indices were compared in infected and uninfected birds. Trematode infection considerably affected host protein metabolism irrespective of the age; however, the changes were more pronounced in nestlings. Increased concentration of γ-globulins, modified albumin, and circulating immune complexes was observed in plasma of infected herring gulls. The experiments demonstrated the most significant changes in protein metabolism of herring gulls 8–11 days after infection with trematode H. larina.     

Real-Time Detection and Identification of Chlamydophila Species in Veterinary Specimens by Using SYBR Green-Based PCR Assays

Infections caused by members of the Chlamydiaceae family have long been underestimated due to the requirement of special laboratory facilities for the detection of this group of intracellular pathogens. Furthermore, new studies of this group of intracellular pathogens have revealed that host specificity of different species is not as clear as recently believed. As most members of the genus Chlamydophila have shown to be transmissible from animals to humans, sensitive and fast detection methods are required. In this study, SYBR green-based real-time assays were developed that detect all members of Chlamydiaceae and differentiate the most prevalent veterinary Chlamydophila species: Cp. psittaci, Cp. abortus, Cp. felis, and Cp. caviae. By adding bovine serum albumin to the master mixes, target DNA could be detected directly in crude lysates of enzymatically digested conjunctival or pharyngeal swabs or tissue specimens from heart, liver, and spleen without further purification. The assays were evaluated on veterinary specimens where all samples were screened using a family-specific PCR, and positive samples were further tested using species-specific PCRs. Cp. psittaci was detected in 47 birds, Cp. felis was found in 10 cats, Cp. caviae was found in one guinea pig, and Cp. abortus was detected in one sheep. The screening assay appeared more sensitive than traditional microscopical examination of stained tissue smears. By combining a fast, robust, and cost-effective method for sample preparation with a highly sensitive family-specific PCR, we were able to screen for Chlamydiaceae in veterinary specimens and confirm the species in positive samples with additional PCR assays.     

Sleep in the herring gull (Larus argentatus)

Sleep postures and eye state of free-ranging herring gulls (Larus argentatus) were studied during the breeding season. Three mutually exclusive behaviours were observed, namely sleep, rest-sleep and rest postures. Arousal thresholds, eye blink rates and eye closure time were obtained during these behaviours. Significant relationships existed between eye blinking, eye closure, and a raised threshold of arousal when birds were in the sleep and rest-sleep postures. During a natural disturbance, birds in the sleep posture remained in this posture but did not blink their eyes: this is called pseudo sleep. Male gulls also exhibited a lower threshold of arousal while in the sleep posture compared with females. We conclude that rhythmic eye blinking is a good indication of sleep in herring gulls.     

Otolith variation in Pacific herring (<Emphasis Type="Italic">Clupea pallasii</Emphasis>) reflects mitogenomic variation rather than the subspecies classification

Pacific herring (Clupea pallasii) is divided into three subspecies: two in northeast Europe and one in the north Pacific Ocean. Genetic studies have indicated that the populations in northeast Europe have derived from the northwest Pacific herring recently, or during the last 10–15 kyr, and that they are distinct from the population in the northeast Pacific. In addition, hybridization between the Pacific herring and the Atlantic herring has been documented. Otolith variation has been considered to be largely affected by environmental variation, but here we evaluate whether the genetic differentiation is reflected in otolith shape differences. A clear difference in otolith shape was observed between the genetically differentiated herring species Clupea harengus from the Atlantic and C. pallasii. The otolith shape of C. p. suworowi in the Barents Sea was different from the shape of C. pallasii in northern Norway and C. p. pallasii from the Pacific. Populations of C. p. pallasii, sampled east and west of the Alaska Peninsula, which belong to two genetically different clades of the C. p. pallasii in the Pacific Ocean, show a clear difference in otolith shape. C. p. suworowi and the local C. pallasii peripheral population in Balsfjord in northern Norway are more similar to the northwest Pacific herring (C. p. pallasii) than to the northeast Pacific herring (C. p. pallasii), both genetically and in otolith shape. The Balsfjord population, known to be influenced by introgression of mtDNA from the Atlantic herring, does not show any sign of admixture in otolith shape between the two species. A revised classification, considering the observed genetic and morphological evidence, should rather group the northwest Pacific herring in the Bering Sea together with the European populations of C. pallasii than with the northeast Pacific herring in the Gulf of Alaska.     

The diet of Atlantic Yellow-legged Gulls (<Emphasis Type="Italic">Larus michahellis atlantis</Emphasis>) at an oceanic seabird colony: estimating predatory impact upon breeding petrels

The diet and breeding ecology of Yellow-legged Gulls (Larus michahellis atlantis) were studied on Selvagem Grande, North Atlantic in the nesting season of 2007. We collected and analyzed 715 pellets from adults. The most frequent prey were White-faced Storm-petrels (Pelagodroma marina; present on 40.8% of all pellets) and the endemic land snails (Theba macandrewiana; present on 36.5% of all pellets). Other birds, namely Cory’s Shearwaters (Calonectris diomedea), Macaronesian Shearwaters (Puffinus assimilis), Bulwer’s Petrels (Bulweria bulwerii), and Band-rumped Storm-petrels (Oceanodroma castro) were relatively less frequent, but overall, seabirds were present in ca. 50% of all pellets, representing an estimated 60.4% of all mass consumed by gulls. We estimate that the contribution of seabirds to the overall caloric balance accounted for 82.5% of all energy consumed. The number of gull pairs breeding on Selvagem Grande was 12 on 2005 and 2007. Breeding success was low (0.92 and 0.25 juveniles per breeding pair, respectively). Using a simple bioenergetics model, we estimate the breeding gull population to have the potential to consume approximately 4,847 adult/sub-adult seabirds in 3.5 months in order to meet its energetic requirements. The importance of the estimated predation levels is discussed and some management actions are suggested.     

An investigation on the presence of <Emphasis Type="Italic">Chlamydiaceae</Emphasis> in Swedish dogs

Background  

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, including man, other mammals, and birds. Upper respiratory and genital diseases are common clinical problems caused by Chlamydiaceae. Very little is known about chlamydial infections in dogs. Few clinical reports on natural disease in dogs describe mainly conjunctival and upper respiratory signs, and the role of Chlamydiaceae in genital disease is unclear. The present study aimed at studying the prevalence of Chlamydiaceae in healthy dogs and in dogs with genital or upper respiratory disease, including conjunctivitis.     

Evidence for the existence of two new members of the family Chlamydiaceae and proposal of Chlamydia avium sp. nov. and Chlamydia gallinacea sp. nov.

The family Chlamydiaceae with the recombined single genus Chlamydia currently comprises nine species, all of which are obligate intracellular organisms distinguished by a unique biphasic developmental cycle. Anecdotal evidence from epidemiological surveys in flocks of poultry, pigeons and psittacine birds have indicated the presence of non-classified chlamydial strains, some of which may act as pathogens. In the present study, phylogenetic analysis of ribosomal RNA and ompA genes, as well as multi-locus sequence analysis of 11 field isolates were conducted. All independent analyses assigned the strains into two different clades of monophyletic origin corresponding to pigeon and psittacine strains or poultry isolates, respectively. Comparative genome analysis involving the type strains of currently accepted Chlamydiaceae species and the designated type strains representing the two new clades confirmed that the latter could be classified into two different species as their average nucleotide identity (ANI) values were always below 94%, both with the closest relative species and between themselves.     

A Real-Time PCR Assay for the Detection of Atypical Strains of Chlamydiaceae from Pigeons

Recent evidence of the occurrence of atypical Chlamydiaceae strains in pigeons, different from the established Chlamydiaceae, requires the development of a specific and rapid detection tool to investigate their prevalence and significance. Here is described a new real-time PCR assay that allows specific detection of atypical Chlamydiaceae from pigeons. The assay has been used to assess the dissemination of these strains in field samples collected from Parisian pigeon populations in 2009. The results suggest a limited dissemination compared to the usually higher prevalence of Chlamydia psittaci that is the main species associated with avian chlamydiosis.     

Nahrungsökologische Untersuchungen an Mantelmöwen(Larus marinus) auf Helgoland

This paper is based on observations and the analysis of 54 stomach contents of great black-backed gulls obtained from the island of Helgoland (North Sea) in 1971 and 1972. It deals with the following problems: How large is the share of food taken from the human area in progressing synanthropy of the great black-backed gull? Does this gull have any negative effect on the colony of guillemots(Uria aalge albionis) on Helgoland? Do great black-backed gulls bring further danger (in addition to weather, etc.) for smaller birds migrating over or resting on Helgoland in spring and autumn? 73.5% of the food taken up is of human origin, only 26.5% is considered to be natural food (Crustacea, Mollusca, Insecta). 36 (66.7%) stomachs contained remainders of fish, mainly cod(Gadus morhua), whiting(Merlangius merlangus) and poor cod(Trisopterus minutus). Together with herring gulls(Larus argentatus), great black-backed gulls often swoop down on fishing guillemots to recover the just captured fish from them. They also attack homing guillemots in the range of their breeding rocks. Only in one case the remainders of a smaller bird were found, but presumably a greater number of smaller birds is killed by great gulls.     

Screening for Viral Hemorrhagic Septicemia Virus in Marine Fish along the Norwegian Coastal Line

Viral hemorrhagic septicemia virus (VHSV) infects a wide range of marine fish species. To study the occurrence of VHSV in wild marine fish populations in Norwegian coastal waters and fjord systems a total of 1927 fish from 39 different species were sampled through 5 research cruises conducted in 2009 to 2011. In total, VHSV was detected by rRT-PCR in twelve samples originating from Atlantic herring (Clupea harengus), haddock (Melanogrammus aeglefinus), whiting (Merlangius merlangus) and silvery pout (Gadiculus argenteus). All fish tested positive in gills while four herring and one silvery pout also tested positive in internal organs. Successful virus isolation in cell culture was only obtained from one pooled Atlantic herring sample which shows that today''s PCR methodology have a much higher sensitivity than cell culture for detection of VHSV. Sequencing revealed that the positive samples belonged to VHSV genotype Ib and phylogenetic analysis shows that the isolate from Atlantic herring and silvery pout are closely related. All positive fish were sampled in the same area in the northern county of Finnmark. This is the first detection of VHSV in Atlantic herring this far north, and to our knowledge the first detection of VHSV in silvery pout. However, low prevalence of VHSV genotype Ib in Atlantic herring and other wild marine fish are well known in other parts of Europe. Earlier there have been a few reports of disease outbreaks in farmed rainbow trout with VHSV of genotype Ib, and our results show that there is a possibility of transfer of VHSV from wild to farmed fish along the Norwegian coast line. The impact of VHSV on wild fish is not well documented.     

Geographical variation in the yellow‐legged gull: introgression or convergence from the herring gull?

Yellow-legged gulls Larus michahellis from the Atlantic Iberian coast exhibit some phenotypic similarities with the herring gull L. argentatus from Western Europe. To assess this phenomenon and its possible origin, we compared Mediterranean yellow-legged gulls, Atlantic Iberian yellow-legged gulls and herring gulls for several phenotypic traits (morphology, plumage), and used genetic data to determine the evolutionary history of the Atlantic Iberian yellow-legged gulls. Data from mitochondrial cytochrome b gene and microsatellite loci clearly indicate that Atlantic Iberian gulls are closely related to Mediterranean yellow-legged gulls, and do not show stronger signs of introgression with herring gulls relative to other populations of yellow-legged gulls. Atlantic Iberian yellow-legged gulls are more similar to herring gulls in body size and shape than to other yellow-legged gulls populations, but not in mantle colour and wing-tip pattern. Body size and other phenotypic and life history similarities with the herring gull ( L. argentatus argenteus ) such as voice, winter plumage and breeding phenology, previously described in several studies, might thus be interpreted as convergent characters. Within the yellow-legged gull, the high F _st-values obtained from four nuclear microsatellite loci indicate substantial population structure and reduced levels of gene flow between gull populations in Mediterranean France and Atlantic Iberia. Differences among these populations in breeding phenology and migration patterns, likely resulting from different local selection pressures, might contribute to this low level of gene flow.     

The activities of digestive enzymes as a determinant factor in the localization of <Emphasis Type="Italic">Tetrabothrius erostris</Emphasis> (Loennberg) (Cestoda: Tetrabothriidae) in the intestine of the herring gull <Emphasis Type="Italic">Larus argentatus</Emphasis> Pontoppidan

We determined the parameters of invasion and localization of Tetrabothrius erostris (Loennberg) (Cestoda: Tetrabothriidae) in the intestine of herring gulls Larus argentatus Pontoppidan (adults and nestlings). Adult herring gulls have T. erostris parasites in the proximal and middle segment of their small intestine. Nestlings have T. erostris in the middle segment of their small intestine. High activities of the digestive enzymes (proteases and glycosidaeses) in these segments of the intestine were found. Apparently, this factor, as well as the physiological peculiarities of the development of T. erostris, determine the localization of tetrabothriids in the intestine of herring gulls.     

Ultraviolet photopigment sensitivity and ocular media transmittance in gulls,with an evolutionary perspective

Gulls (Laridae excluding Sternidae) appear to be the only shorebirds (Charadriiformes) that have a short wavelength sensitive type 1 (SWS1) cone pigment opsin tuned to ultraviolet (UV) instead of violet. However, the apparent UV-sensitivity has only been inferred indirectly, via the interpretation that the presence of cysteine at the key amino acid position 90 in the SWS1 opsin confers UV sensitivity. Unless the cornea and the lens efficiently transmit UV to the retina, gulls might in effect be similar to violet-sensitive birds in spectral sensitivity even if they have an ultraviolet sensitive (UVS) SWS1 visual pigment. We report that the spectral transmission of the cornea and lens of great black-backed Larus marinus and herring gulls L. argentatus allow UV-sensitivity, having a λ_T0.5 value, 344 nm, similar to the ocular media of UV sensitive birds. By molecular sequencing of the second α-helical transmembrane region of the SWS1 opsin gene we could also infer that 15 herring gulls and 16 yellow-legged gulls L. michahellis, all base-pair identical, are genetically UV-sensitive.     

Assessment of Chlamydia psittaci Shedding and Environmental Contamination as Potential Sources of Worker Exposure throughout the Mule Duck Breeding Process

Chlamydia psittaci is an obligate intracellular bacterium responsible for avian chlamydiosis, otherwise known as psittacosis, a zoonotic disease that may lead to severe atypical pneumonia. This study was conducted on seven mule duck flocks harboring asymptomatic birds to explore the circulation and persistence of C. psittaci during the entire breeding process and assess the potential sources of worker exposure. Cloacal swabs and air samples were taken on each occasion requiring humans to handle the birds. In parallel, environmental samples, including dust, water, and soil, were collected. Specific real-time PCR analyses revealed the presence of C. psittaci in all flocks but with three different shedding patterns involving ducks about the age of 4, 8, and 12 weeks with heavy, moderate, and low excretion levels, respectively. Air samples were only positive in flocks harboring heavy shedders. Dust in flocks with heavy or moderate shedders carried chlamydial loads strongly associated with the loads detected in avian and soil samples. Environmental contamination, significantly correlated with shedding dynamics, was considered to be the most probable source of exposure. The high prevalence of bacteriophage Chp1 in all flocks, mostly jointly present with chlamydia, suggests an important factor in C. psittaci persistence, thus creating a greater risk for humans. A survey conducted in these flocks regarding farming practices and activities showed that disinfection seems to be the most promising practice for reducing C. psittaci prevalence in ducks and that the place and the duration of action during operations seem to be potential risk factors. Strict adherence to good practices is strongly recommended.     

Construction of Recombinant HVT Expressing PmpD,and Immunological Evaluation against Chlamydia psittaci and Marek’s Disease Virus

Chlamydia psittaci (C. psittaci) is an obligate intracellular zoonotic pathogen that can be transmitted to humans from birds. No efficacious commercial vaccine is available for clearing chlamydial infection due to lack of potential vaccine candidates and effective delivery vehicles. Herpesvirus of turkeys (HVT) is an efficacious commercially available vaccine against Marek’s Disease virus (MDV). In this study, a recombinant HVT-delivered vaccine against C. psittaci and Marek’s disease was developed and examined. The 5''-terminus of pmpD gene (pmpD-N) encoding the N-terminal fragment of polymorphic membrane protein D of C. psittaci was inserted into a nonessential region of HVT genome using reverse genetics based on an infectious bacterial artificial chromosome (BAC) clone of HVT. The recombinant virus (rHVT-pmpD-N) was recovered from primary chicken embryo fibroblast (CEF) cells by transfection of modified HVT BAC DNA containing the pmpD-N gene. The rHVT-pmpD-N construct was confirmed to express PmpD-N by immunoblot and immunofluorescence. The rHVT-pmpD-N was stable during 20 passages in vitro. The growth kinetics of rHVT-pmpD-N was comparable to that of parental HVT in vitro and in vivo. One-day-old SPF chickens inoculated subcutaneously with rHVT-pmpD-N displayed increased PmpD-specific antibody levels and a vigorous PmpD-specific lymphocyte proliferation response using HVT vector or CEF cells as control. Furthermore, the percentage of CD4+ cells was significantly elevated in rHVT-pmpD-N-immunized birds as compared to the parental HVT. All chickens vaccinated with rHVT-pmpD-N or parental HVT were protected completely against challenge with a very virulent strain of Marek’s Disease virus (MDV) RB-1B. Post challenge with C. psittaci CB7 strain, a significant decrease in respiratory distress, lesions and Chlamydia load was found in the rHVT-pmpD-N-vaccinated group compared to the parental HVT. In conclusion, our study suggests that the rHVT-pmpD-N live vaccine may be viable as a candidate dual vaccine that provides protection against both very virulent MDV and C. psittaci.