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Avian trichomonosis is an upper digestive tract disease of birds typically caused by the protozoan parasite Trichomonas gallinae. In California (U.S.A), trichomonosis is known to cause periodic epidemics in the Pacific Coast band-tailed pigeon (Patagioenas fasciata monolis), a migratory upland game bird. We summarize the mortality events that occurred during winter 2014-2015 including the duration, estimated mortality, pathology, and genetic identity of infecting parasites. Increased mortality was reported from locations in 25 counties between November 2014 and June 2015. Based on reports, carcasses received, wildlife rehabilitation center admissions, site visits, and regular monitoring by local personnel, total mortality was estimated at 18,440. At necropsy, birds had multiple coalescing lesions in the oral cavity involving the upper palate and/or around the tongue and glottis, esophagus, crop, and/or proventriculus. Birds collected from Contra Costa (63.9%; 30/47); Marin (75.0%; 6/8), San Mateo (46.7%; 14/30), and Santa Clara (35.0%; 37/106) counties were more likely to have lesions extending into their head involving muscle, sinuses, ear canals, eye sockets, and bone (χ2 = 62.9; df = 11; P < 0.001). Histopathologic findings included pharyngitis, esophagitis, myositis, and air sacculitis of the pneumatic bone of the skull. Mixed bacterial colonies were found multifocally at the fronts of the necrosis in six of the eleven birds examined histologically. Infecting trichomonads included T. gallinae subtype A2 (n = 5), un-typed T. gallinae (n = 4), mixed infection with T. gallinae subtype A2 and T. stableri (n = 1), and mixed infection with un-typed T. gallinae and T. stableri (n = 1). The winter 2014-2015 epidemic was the largest on record in terms of duration, locations, and birds affected. Infection dynamics may have been exacerbated by the drought in California. Increased monitoring of band-tailed pigeons is needed to understand the long-term impacts of large-scale mortality events on their population.

Avian trichomonosis, caused by the flagellated protozoan parasite Trichomonas gallinae, has variable pathogenicity among bird species ranging from asymptomatic infections to severe disease periodically manifesting in epidemic mortality. Traditionally, columbids are identified as highly susceptible to infection with occasional spillover into raptors that prey on infected birds. We identified avian trichomonosis in two dead California spotted owls (Strix occidentalis occidentalis) and three dead northern spotted owls (S. o. caurina) in California during 2011-2015; infection was confirmed in four owls by PCR. Pathologic lesions associated with trichomonosis in the owls included caseonecrotic lesions of the upper palate accompanied by oropharyngitis, cellulitis, myositis, and/or sinusitis. Spotted owls are known to mainly feed on small mammals; therefore, the source of infection as well as the significance of the disease in spotted owls is unclear. These owl trichomonosis cases coincided temporally and spatially with three trichomonosis epidemics in band-tailed pigeons (Patagioenas fasciata monilis). The same parasite, T. gallinae subtype A2, was isolated from the spotted owls and band-tailed pigeons, suggesting the owls became infected when opportunistically feeding on pigeons during mortality events. Avian trichomonosis is an important factor in the decline of the Pacific Coast band-tailed pigeon population with near-annual mortality events during the last 10 years and could have conservation implications for raptor species at risk, particularly those that are facing multiple threats.

The Pacific Coast band-tailed pigeon (Patagioenas fasciata monilis) is a migratory game bird of North America that is at risk for population decline. Epidemics of avian trichomonosis caused by upper digestive tract infection with Trichomonas spp. protozoa in these and other doves and pigeons of the United States are sporadic, but can involve tens of thousands of birds in a single event. Herein, we analyze the role of trichomonosis in band-tailed pigeon mortality and relate spatial, temporal and demographic patterns of parasite transmission to the genetic background of the infecting organism. Infections were most common in adult birds and prevalence was high in band-tailed pigeons sampled at mortality events (96%) and rehabilitation centers (36%) compared to those that were hunter-killed (11%) or live-caught (4%). During non-epidemic periods, animals were primarily infected with T. gallinae Fe-hydrogenase subtype A2, and were less often infected with either T. gallinae subtype A1 (the British finch epidemic strain), T. stableri n. sp. (a T. vaginalis-like species), or Tritrichomonas blagburni n. sp.-like organisms. Birds sampled during multiple epidemics in California were only infected with T. gallinae subtype A2 and T. stableri. The non-clonal etiology of avian trichomonosis outbreaks in band-tailed pigeons and the risk of spill-over to raptor and passerine species highlights the need for additional studies that clarify the host range and evolutionary relationships between strains of Trichomonas spp. in regions of trichomonosis endemicity.

Trichomonas gallinae is a ubiquitous flagellated protozoan parasite, and the most common etiologic agent of epidemic trichomonosis in columbid and passerine species. In this study, free-ranging Pacific Coast band-tailed pigeons (Patagioenas fasciata monilis) in California (USA) were found to be infected with trichomonad protozoa that were genetically and morphologically distinct from T. gallinae. In microscopic analysis, protozoa were significantly smaller in length and width than T. gallinae and were bimodal in morphology. Phylogenetic analysis of the ITS1/5.8S/ITS2, rpb1, and hydrogenosomal Fe-hydrogenase regions revealed that the protozoan shares an ancestor with Trichomonas vaginalis, the sexually-transmitted agent of trichomoniasis in humans. Clinical and pathologic features of infected birds were similar to infections with T. gallinae. Evidence presented here strongly support taxonomical distinction of this parasite, which we hereby name Trichomonas stableri n. sp. This work contributes to a growing body of evidence that T. gallinae is not the sole etiologic agent of avian trichomonosis, and that the incorporation of molecular tools is critical in the investigation of infectious causes of mortality in birds.

Avian trichomonosis, caused by the flagellated protozoan parasite Trichomonas gallinae, has variable pathogenicity among bird species ranging from asymptomatic infections to severe disease periodically manifesting in epidemic mortality. Traditionally, columbids are identified as highly susceptible to infection with occasional spillover into raptors that prey on infected birds. We identified avian trichomonosis in two dead California spotted owls (Strix occidentalis occidentalis) and three dead northern spotted owls (S. o. caurina) in California during 2011–2015; infection was confirmed in four owls by PCR. Pathologic lesions associated with trichomonosis in the owls included caseonecrotic lesions of the upper palate accompanied by oropharyngitis, cellulitis, myositis, and/or sinusitis. Spotted owls are known to mainly feed on small mammals; therefore, the source of infection as well as the significance of the disease in spotted owls is unclear. These owl trichomonosis cases coincided temporally and spatially with three trichomonosis epidemics in band-tailed pigeons (Patagioenas fasciata monilis). The same parasite, T. gallinae subtype A2, was isolated from the spotted owls and band-tailed pigeons, suggesting the owls became infected when opportunistically feeding on pigeons during mortality events. Avian trichomonosis is an important factor in the decline of the Pacific Coast band-tailed pigeon population with near-annual mortality events during the last 10 years and could have conservation implications for raptor species at risk, particularly those that are facing multiple threats.

Avian trichomonosis is an architypal disease of wild columbids and those birds that predate them. Increasingly though, it has been reported in passerines; a recent and ongoing epidemic in the chaffinches and greenfinches of Europe and outbreaks amongst house finches, American goldfinches and purple finches in North America. The parasite, Trichomonas gallinae , causes lesions in the upper respiratory tract which can cause mortality associated with dehydration and emaciation. This paper reports for the first time, the widespread, endemic and often asymptomatic infection of common mynah (Acridotheres tristis) around the Faisalabad District, Pakistan. Parasite typing was used to investigate the potential for transmission among the frequently sympatric species. Type C parasites were found in mynah, and while this is analagous to the pandemic finch strain which is Type A, it is the first known example of passerine infections of this parasite genotype. Subtype analysis showed the strain to be C4 a subtype which has a widespread distribution in columbids. [ABSTRACT FROM AUTHOR]

Mono‐iron hydrogenase ([Fe]‐hydrogenase) reversibly catalyzes the transfer of a hydride ion from H2 to methenyltetrahydromethanopterin (methenyl‐H4MPT+) to form methylene‐H4MPT. Its iron guanylylpyridinol (FeGP) cofactor plays a key role in H2 activation. Evidence is presented for O2 sensitivity of [Fe]‐hydrogenase under turnover conditions in the presence of reducing substrates, methylene‐H4MPT or methenyl‐H4MPT+/H2. Only then, H2O2 is generated, which decomposes the FeGP cofactor; as demonstrated by spectroscopic analyses and the crystal structure of the deactivated enzyme. O2 reduction to H2O2 requires a reductant, which can be a catalytic intermediate transiently formed during the [Fe]‐hydrogenase reaction. The most probable candidate is an iron hydride species; its presence has already been predicted by theoretical studies of the catalytic reaction. The findings support predictions because the same type of reduction reaction is described for ruthenium hydride complexes that hydrogenate polar compounds. [ABSTRACT FROM AUTHOR]

Histomonas meleagridis, a protozoan parasite that can infect gallinaceous birds, affects mainly the liver and caeca of infected birds. As a consequence of the recent ban of chemotherapeuticals in Europe and the USA, histomonosis gained somewhat more attention due to its re-emergence and the fact that there is no effective treatment available. Therefore, special attention is now also given towards the diagnosis and the control of the disease. In the actual study we report the development of highly specific and sensitive real-time PCR methods for detection and quantification of the parasite, based on two protein coding genes, Fe-hydrogenase ( FeHYD ) and rpb1 . Both genes seem to be in a single copy in H. meleagridis as shown by southern blotting and absolute quantification using real-time PCRs on samples containing a known amount of the parasite. The real-time PCR assays based on FeHYD and rpb1 genes were found to be an efficient method for the quantification and detection of H. meleagridis in in vitro grown cultures, tissues of infected birds and in faecal samples. Both real-time PCRs were able to detect up to a single cell in in vitro cultures of H. meleagridis and in fecal samples spiked with H. meleagridis . Finally, qPCR assays were shown to be highly specific for H. meleagridis as samples containing either of the two H. meleagridis genotypes were positive, whereas samples containing other protozoa such as Tetratrichomonas gallinarum, Trichomonas gallinae, Simplicimonas sp., Tritrichomonas sp., Parahistomonas wenrichi , Dientamoebidae sp. and Blastocystis sp. were all negative. [ABSTRACT FROM AUTHOR]

[Fe]-hydrogenase (Hmd), an enzyme of the methanogenic energy metabolism, harbors an iron-guanylylpyridinol (Fe GP) cofactor used for H2 cleavage. The generated hydride is transferred to methenyl-tetrahydromethanopterin (methenyl-H4 MPT+). Most hydrogenotrophic methanogens contain the hmd-related genes hmd II and hmd III. Their function is still elusive. We were able to reconstitute the Hmd II holoenzyme of Methanocaldococcus jannaschii with recombinantly produced apoenzyme and the Fe GP cofactor, which is a prerequisite for in vitro functional analysis. Infrared spectroscopic and X-ray structural data clearly indicated binding of the Fe GP cofactor. Methylene-H4 MPT binding was detectable in the significantly altered infrared spectra of the Hmd II holoenzyme and in the Hmd II apoenzyme-methylene-H4 MPT complex structure. The related binding mode of the Fe GP cofactor and methenyl-H4 MPT+ compared with Hmd and their multiple contacts to the polypeptide highly suggest a biological role in Hmd II. However, holo-Hmd II did not catalyze the Hmd reaction, not even in a single turnover process, as demonstrated by kinetic measurements. The found inactivity can be rationalized by an increased contact area between the C- and N-terminal folding units in Hmd II compared with in Hmd, which impairs the catalytically necessary open-to-close transition, and by an exchange of a crucial histidine to a tyrosine. Mainly based on the presented data, a function of Hmd II as Hmd isoenzyme, H2 sensor, Fe GP-cofactor storage protein and scaffold protein for Fe GP-cofactor biosynthesis could be excluded. Inspired by the recently found binding of Hmd II to aminoacyl- tRNA synthetases and tRNA, we tentatively consider Hmd II as a regulatory protein for protein synthesis that senses the intracellular methylene-H4 MPT concentration. Database Structural data are available in the Protein Data Bank under the accession numbers ; ; and . [ABSTRACT FROM AUTHOR]

Finch trichomonosis, caused by Trichomonas gallinae, emerged in the Canadian Maritime provinces in 2007 and has since caused ongoing mortality in regional purple finch (Carpodacus purpureus) and American goldfinch (Carduelis tristis) populations. Trichomonas gallinae was isolated from (1) finches and rock pigeons (Columbia livia) submitted for post-mortem or live-captured at bird feeding sites experiencing trichomonosis mortality; (2) bird seed at these same sites; and (3) rock pigeons live-captured at known roosts or humanely killed. Isolates were characterized using internal transcribed spacer (ITS) region and iron hydrogenase (Fe-hyd) gene sequences. Two distinct ITS types were found. Type A was identical to the UK finch epidemic strain and was isolated from finches and a rock pigeon with trichomonosis; apparently healthy rock pigeons and finches; and bird seed at an outbreak site. Type B was obtained from apparently healthy rock pigeons. Fe-hyd sequencing revealed six distinct subtypes. The predominant subtype in both finches and the rock pigeon with trichomonosis was identical to the UK finch epidemic strain A1. Single nucleotide polymorphisms in Fe-hyd sequences suggest there is fine-scale variation amongst isolates and that finch trichomonosis emergence in this region may not have been caused by a single spill-over event. [ABSTRACT FROM AUTHOR]

Highlights: [•] Pacific Coast band-tailed pigeons were tested for trichomonad infection and disease. [•] Protozoa were genetically characterized at ITS1/5.8S/ITS2 and Fe-hydrogenase loci. [•] Trichomonas gallinae Fe-hydrogenase subtype A2 was the dominant pathogenic genotype. [•] Both T. gallinae and T. stableri were isolated at large trichomonosis outbreaks. [ABSTRACT FROM AUTHOR]

SUMMARY: In recent years, Trichomonas gallinae emerged as the causative agent of an infectious disease of passerine birds in Europe leading to epidemic mortality of especially greenfinches Chloris chloris and chaffinches Fringilla coelebs. After the appearance of finch trichomonosis in the UK and Fennoscandia, the disease spread to Central Europe. Finch trichomonosis first reached Austria and Slovenia in 2012. In the present study the genetic heterogeneity of T. gallinae isolates from incidents in Austria and Slovenia were investigated and compared with British isolates. For this purpose comparative sequence analyses of the four genomic loci ITS1-5.8S-ITS2, 18S rRNA, rpb1 and Fe-hydrogenase were performed. The results corroborate that one clonal T. gallinae strain caused the emerging infectious disease within passerine birds and that the disease is continuing to spread in Europe. The same clonal strain was also found in a columbid bird from Austria. Additionally, the present study demonstrates clearly the importance of multi-locus sequence typing for discrimination of circulating T. gallinae strains. [ABSTRACT FROM AUTHOR]

SUMMARY: Avian trichomonosis, caused by the flagellated protozoan Trichomonas gallinae , is a recently emerged infectious disease of British passerines. The aetiological agent, a clonal epidemic strain of the parasite, has caused unprecedented finch mortality and population-level declines in Britain and has since spread to continental Europe. To better understand the potential origin of this epidemic and to further investigate its host range, T. gallinae DNA extracts were collected from parasite culture and tissue samples from a range of avian species in Britain. Sequence typing at the ITS1/5.8S rRNA/ITS2 region resolved three distinct ITS region types circulating in free-ranging British birds. Subtyping by sequence analyses at the Fe-hydrogenase gene demonstrated further strain variation within these ITS region types. The UK finch epidemic strain was preponderant amongst columbids sampled, however, wide strain diversity was encountered in isolates from a relatively small number of pigeons, suggesting further strains present in columbid populations across the UK are yet to be identified. Fe-hydrogenase gene sequence data in isolates from birds of prey with disease were predominantly identical to the UK finch epidemic strain, demonstrating its presence as a virulent strain in UK birds of prey since at least 2009. [ABSTRACT FROM AUTHOR]

Abstract: Fe-hydrogenase from Enterobacter cloacae IIT-BT08 was purified 1284 fold with specific activity of 335μmol H2/min/mg protein for hydrogen evolution using reduced methyl viologen as an electron-donor at 25°C. The molecular weight of the monomeric enzyme was determined to be 51kDa by MALDI-ToF mass spectrometry. The PI of the enzyme was ∼5.6 displaying its acidic nature. The optimal temperature and pH for hydrogen evolution was 37°C and 7–7.2 respectively. The affinity constant, Km for reduced methyl viologen was 0.57±0.03mM and that of reduced ferredoxin was 0.72±0.04μM. The enzyme contained ∼11.47gm-atom Fe/mol of Fe-hydrogenase. Electron paramagnetic resonance analysis ascertained the existence of iron molecules as [4Fe–4S] clusters. The internal amino acid sequences of trypsin digested peptides of hydrogenase as determined by ESI MS/MS Q-ToF showed 80-87% identities with the respective sequences of Clostridium sp. and Trichomonas sp. hydrogenase. [Copyright &y& Elsevier]

Abstract: Biological H2 production has an edge over its chemical counterpart mainly because it is environmentally benign. Despite having simpler technology, higher evolution rate of H2 and the wide spectrum of substrate utilization, the major deterrent of anaerobic dark fermentation process stems from its lower achievable yields. Theoretically, the maximum H2 yield is 4molH2/molglucose when glucose is completely metabolized to acetate or acetone in the anaerobic process. But it is somewhat difficult to achieve the complete degradation of glucose to carbon dioxide and H2 through anaerobic dark fermentation. Moreover, this yield appears too low to be economically viable as an alternative to the existing chemical or electrochemical processes of hydrogen generation. Intensive research studies have already been carried out on the advancement of these processes, such as the development of genetically modified microorganism, improvement of the reactor designs, use of different solid matrices for the immobilization of whole cells, development of two-stage processes, and higher H2 production rates. Maximum H2 yield is found to be 5.1molH2/molglucose. However, major bottlenecks for the commercialization of these processes are lower H2 yield and rate of H2 production. Competent microbial cultures are required to handle waste materials efficiently, which are usually complex in nature. This will serve dual purposes: clean energy generation and bioremediation. Scale-up studies on fermentative H2 production processes have been done successfully. Pilot plant trials of the photo-fermentation processes require more attention. Use of cheaper raw materials and efficient biological H2 production processes will surely make them more competitive with the conventional H2 generation processes in near future. [Copyright &y& Elsevier]

Degenerate PCR primers (forward as well as the reverse) were designed from the protein sequences of known structural genes encoding the catalytic subunits of Fe-hydrogenases obtained from NCBI Protein Sequence Database. Amino acid sequences were multiple aligned through CLUSTAL-W software and compared for conserved sequence motifs. Mole percent of G+C of the present strain is found to be 55% by thermal melting method. An amplified PCR product of 1 kb in size was obtained from the genomic DNA of Enterobacter cloacae IIT-BT 08 by using a set of (JM-1 and JM-12) degenerate primer. Gel purified PCR product was cloned in TOPO TA cloning vector (3.9 kb) and transformed into TOPOF’ cells. Initial experiment on Southern hybridization of the PCR product showed no homology with hydA gene of Clostridium acetobutylicum ATCC 824. [Copyright &y& Elsevier]

Chlamydomonas reinhardtii , a unicellular green alga, contains a hydrogenase enzyme, which is induced by anaerobic adaptation of the cells. Using the suppression subtractive hybridization (SSH) approach, the differential expression of genes under anaerobiosis was analyzed. A␣PCR fragment with similarity to the genes of bacterial Fe-hydrogenases was isolated and used to screen an anaerobic cDNA expression library of C. reinhardtii . The cDNA sequence of hydA contains a 1494-bp ORF encoding a protein with an apparent molecular mass of 53.1 kDa. The transcription of the hydrogenase gene is very rapidly induced during anaerobic adaptation of the cells. The deduced amino-acid sequence corresponds to␣two␣polypeptide sequences determined by sequence analysis of the isolated native protein. The Fe-hydrogenase contains a short transit peptide of 56 amino acids, which routes the hydrogenase to the chloroplast stroma. The isolated protein belongs to a new class of Fe-hydrogenases. All four cysteine residues and 12 other␣amino acids, which are strictly conserved in the active site (H-cluster) of Fe-hydrogenases, have been identified. The N-terminus of the C. reinhardtii protein is markedly truncated compared to other nonalgal Fe-hydrogenases. Further conserved cysteines that coordinate additional Fe–S-cluster in other Fe-hydrogenases are missing. Ferredoxin PetF, the natural electron donor, links the hydrogenase from C. reinhardtii to the photosynthetic electron transport chain. The hydrogenase enables the survival of the green algae under anaerobic conditions by transferring the electrons from reducing equivalents to the enzyme. [ABSTRACT FROM AUTHOR]

Trichomonas gallinae causes avian oropharyngeal trichomonosis. This pathogen affects a large number of bird species and may cause substantial economic losses to the poultry industry. Al-Azizia poultry market in Riyadh, Saudi Arabia is among the largest poultry markets in the Arabian Gulf. Birds traded in this market may be exposed to a variety of T. gallinae strains. Genetic diversity of T. gallinae among birds in the market was examined using Fe‑hydrogenase gene sequences. These sequences were amplified by PCR for twenty-nine isolates of T. gallinae from four different avian species, including 21 feral pigeons, one common mynah, three chickens, and four turkeys. Sequence analysis showed ten variant gene sequences. Nine sequences comprise a new subtype, including A(KSAF1), C(KSAF1) and C(KSAF3) with 34.48% (n = 10), 6.90% (n = 2), 6.90% (n = 2) of the isolates, respectively. Analyses also showed an additional five new sequences (KSAF1.1., KSAF2, KSAF13, KSAF14, KSAF15), representing 17.24% of the isolates. Subtype II (KSAF) was found in four feral pigeons (13.80%). To our knowledge, this report is the first to describe genotypes of T. gallinae from pigeons in Saudi Arabia using Fe‑hydrogenase gene sequences for subtyping. Subtype analysis infers the presence of multiple genotypes of T. gallinae in Saudi avian populations. • Identification of genetic variation within T. gallinae from different avian species in Riyadh, Saudi Arabia using PCR. • This study is the first to describe genotypes of T. gallinae from birds in Riyadh using Fe-hydrogenase gene for subtyping. • Nine novel subtypes of Trichomonas gallinae using Fe-hydrogenase gene as the subtyping locus are described. [ABSTRACT FROM AUTHOR]