A critically co-endangered feather louse Forficuloecus pezopori n. sp. (Phthiraptera: Philopteridae) detected through conservation intervention for the western ground parrot Pezoporus flaviventris (Psittaculidae).

Forficuloecus pezopori Martin, Keatley & Ash n. sp. from the western ground parrot Pezoporus flaviventris North, 1911 (Psittaculidae) is proposed based on combined evidence from morphology and COI mitochondrial DNA. Phylogenetically, the new species is closest to its two known congeners from Western Australia: F. josephi Price, Johnson & Palma, 2008 from Bourke's parrot Neopsephotus bourkii (Gould, 1841) and the scarlet-chested parrot Neophema splendida (Gould, 1841), and F. palmai Guimarães, 1985 from the Australian ringneck parrot Barnardius zonarius (Shaw, 1805). Morphologically it is distinguishable by abdominal chaetotaxy and characters of the male genitalia, and is most similar to F. josephi and F. greeni Guimarães, 1985; the latter has no representative sequence data. Forficuloecus pezopori is the eleventh species of its genus and the only metazoan parasite known from P. flaviventris, which is among Australia's most endangered vertebrates. The new louse is apparently restricted to P. flaviventris and is therefore co-endangered, facing at least the same likelihood of extinction as its host. We recommend ongoing translocation and field monitoring efforts for P. flaviventris include monitoring but not treatment for lice infestations in otherwise healthy individuals, and that the care management plan for captive P. flaviventris considers that F. pezopori is similarly imperilled.

The challenges of detecting Taenia solium and neurocysticercosis in low and middle-income countries: A scoping review of Lao People's Democratic Republic.

OBJECTIVE: Taenia solium is a tapeworm of global importance due to the burden of disease associated with human epilepsy caused by neurocysticercosis. Unfortunately, diagnostic challenges impede control efforts in many low and middle-income countries. This review examines publications concerning Taenia species in the Lao PDR with a focus on T. solium to inform future research and control programmes. METHODS: PubMed and Scopus databases were primary sources of evidence. Publications must have reported taeniasis or T. solium results from Lao PDR. Publications repeating results or samples were combined into unique projects. RESULTS: A total of 64 publications were included and summarised into 46 projects. The majority of projects applied faecal microscopy as their only diagnostic technique. As a result, the specific species of Taenia was often not determined. Only five projects used molecular techniques to identify the species observed. Only case report of neurocysticercosis has been published. The northern region was included in half the number of projects compared to the south, despite being a high-risk area T. solium. CONCLUSIONS: The diagnostic challenge of determining the species of Taenia present in a faecal sample presents a significant limitation to the control of T. solium in Lao PDR and is an experience that is relevant to many other low and middle-income countries. There must be an improved understanding of the distribution and frequency of T. solium if disease control efforts are to be intensified to reduce the burden of neurocysticercosis, as encouraged by WHO and others. It is hoped that this can be achieved through non-biological risk mapping tools and the more frequent application of molecular tools to routine sample collection. Diagnostic tools that are applicable to low resource settings should be a priority area of research for T. solium.

Morphological and Molecular Characterisation of a New Species of Parastrongyloides (Rhabditida: Strongyloididae) from the Short-Beaked Echidna Tachyglossus aculeatus from Western Australia.

INTRODUCTION: A new species of the nematode genus Parastrongyloides Morgan, 1928 was found in the caecum of six short-beaked echidnas Tachyglossus aculeatus (Shaw, 1792) collected from southwestern Australia between August 1964 and March 2020. METHODS: Specimens were prepared for microscopic examination as temporary wet mounts, measurements were made using an Olympus DP71 camera with cellSens standard software, figures were drawn using a drawing tube and light micrographs taken. DNA was extracted using a Qiagen blood and tissue kit, amplified targeting the COX1 gene region. Sequences obtained were analysed and edited using Geneious v.8.1 and aligned to existing sequences published in Genbank using MUSCLE. RESULTS: Parastrongyloides spratti n. sp. can be distinguished from all other species of Parastrongyloides in having the male caudal papillae arranged as a single median dome-shaped pre-cloacal papilla, three tiny pairs of ventral papillae immediately pre-cloacal, a tiny ventral pair of papillae post-cloacal and the female with four to five pairs of dorsoventral papillae immediately anterior to the vulva. A revised key to the species of Parastrongyloides found in Australia is given. Sequence analysis of the COX1 gene corroborated the species status of P. spratti. DISCUSSION: Morphological and molecular analyses support the status of P. spratti as a new species. Parastrongyloides may have an ancient origin in the Australian portion of Gondwanaland.

Molecular Detection of Trypanosoma spp. in Questing and Feeding Ticks (Ixodidae) Collected from an Endemic Region of South-West Australia.

A growing number of indigenous trypanosomes have been reported to naturally infect a variety of Australian wildlife with some species of Trypanosoma implicated in the population decline of critically endangered marsupials. However, the mode of transmission of Australian trypanosomes is unknown since their vectors remain unidentified. Here we aimed to fill this current knowledge gap about the occurrence and identity of indigenous trypanosomes in Australian invertebrates by conducting molecular screening for the presence of Trypanosoma spp. in native ticks collected from south-west Australia. A total of 231 ticks (148 collected from vegetation and 83 retrieved directly from 76 marsupial hosts) were screened for Trypanosoma using a High-Resolution Melt (HRM) qPCR assay. An overall Trypanosoma qPCR positivity of 37% (46/125) and 34% (26/76) was detected in questing ticks and host-collected (i.e., feeding) ticks, respectively. Of these, sequencing revealed 28% (35/125) of questing and 28% (21/76) of feeding ticks were infected with one or more of the five species of trypanosome previously reported in this region (T. copemani, T. noyesi, T. vegrandis, T. gilletti, Trypanosoma sp. ANU2). This work has confirmed that Australian ticks are capable of harbouring several species of indigenous trypanosome and likely serve as their vectors.

'Hook, line, and sinker': Fluorescence in situ hybridisation (FISH) uncovers Trypanosoma noyesi in Australian questing ticks.

Trypanosomes are blood-borne parasites infecting a range of mammalian hosts worldwide. In Australia, an increasing number of novel Trypanosoma species have been identified from various wildlife hosts, some of which are critically endangered. Trypanosoma noyesi is a recently described species of biosecurity concern, due to a close relationship to the South American human pathogen, Trypanosoma cruzi. This genetic similarity increases the risk for introduction of T. cruzi via a local vector. Unfortunately, there is a lack of knowledge concerning the vectorial capacity of Australian invertebrates for native Trypanosoma species. Australian ixodid ticks (Ixodidae), which are widespread ectoparasites of mammalian wildlife, have received the most attention as likely candidates for trypanosome transmission and have been previously implicated as vectors. However, as all studies to date have focused on blood-fed ticks collected directly from infected mammalian hosts, the question of whether ticks maintain a trypanosome infection between blood meals is unknown. In this study, we investigated the presence of Trypanosoma within 148 Australian adult and nymph questing ticks of the species Amblyomma triguttatum, Ixodes australiensis, Ixodes myrmecobii and larvae Ixodes spp., collected from an endemic region of south-west Australia. Using a novel HRM-qPCR detection method that can discriminate between species of Trypanosoma based on primer melting temperature (Tm), we report the first molecular detection of Trypanosoma DNA in Australian questing ticks, with 6 ticks DNA positive for T. noyesi. Additionally, the presence of intact T. noyesi parasites within all (n = 3) smeared gut and gland contents of questing ticks was confirmed using a fluorescence in situ hybridisation (FISH) assay. Whilst this study was unable to determine the in situ tissue location of trypanosomes for the purpose of discerning a potential route of transmission, these combined molecular and FISH smear data indicate that trypanosomes can persist in ticks between blood meals and that ticks are possibly vectors in the transmission of T. noyesi between native wildlife. Transmission experiments are still required to evaluate the competency of Australian ticks as vectors for T. noyesi. Nevertheless, these novel findings warrant further investigation concerning potential life stages and the development of trypanosomes in both Australian, and other, tick species.

Identification of a novel species of Eimeria Schneider, 1875 from the woylie, Bettongia penicillata Gray (Diprotodontia: Potoroidae) and the genetic characterisation of three Eimeria spp. from other potoroid marsupials.

Faecal samples (n = 1,093) collected from the woylie Bettongia penicillata Gray, in south-western Australia were examined for the presence of coccidian parasites. Eimeria sp. oöcysts were detected in 15.2% of samples. Faecal samples obtained from the eastern bettong Bettongia gaimardi (Desmarest) (n = 4) and long-nosed potoroo Potorous tridactylus (Kerr) (n = 12) in Tasmania, were also screened for the presence of Eimeria spp. (prevalence 50% and 41.7%, respectively). Morphological and genetic comparison with other known species of Eimeria indicates that the material identified in woylies is novel. This study aimed to (i) morphologically describe and genetically characterise Eimeria woyliei n. sp. found in woylies; and (ii) genetically characterise Eimeria gaimardi Barker, O'Callaghan & Beveridge, 1988, Eimeria potoroi Barker, O'Callaghan & Beveridge, 1988, and Eimeria mundayi Barker, O'Callaghan & Beveridge, 1988, from other potoroid marsupials. Molecular phylogenetic analyses conducted at the 18S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) loci revealed that E. woyliei n. sp. was most closely related to Eimeria setonicis Barker, O'Callaghan & Beveridge, 1988, at the 18S rDNA locus, and Eimeria trichosuri O'Callaghan & O'Donoghue, 2001, at the cox1 locus. Eimeria woyliei n. sp. is the sixth species of Eimeria to be formally described from potoroid marsupials.

Altered parasite community structure in an endangered marsupial following translocation.

Fauna translocations play an integral role in the management of threatened wildlife, though we are limited by our understanding of how the host-parasite community changes during translocation. During this longitudinal field-based study, we monitored gastrointestinal, blood-borne and ectoparasite taxa infecting woylies (Bettongia penicillata) for up to 12 months following two fauna translocations to supplement existing wild woylie populations in three different sites (Dryandra, Walcott and Warrup East) within the south-west of Western Australia. We aimed to (a) identify changes in parasite community structure of both translocated and resident woylies following translocation; and (b) evaluate the efficacy of ivermectin treatment in translocated hosts. Destination site and time since translocation had the strongest effects on parasite prevalence and mean faecal egg counts following translocation. Ivermectin treatment did not significantly reduce parasite prevalence or mean faecal egg counts in treated hosts. Prior to translocation, parasite community composition differed significantly between woylies selected for translocation and resident woylies within each release site. Following translocation, the parasite communities of translocated and resident hosts converged to become more similar over time, with loss of parasite taxa and novel host-parasite associations emerging. This is the first study to examine changes to the broader parasite community in translocated and resident animals following translocation. The dominant site-specific response of parasites following translocation reinforces the importance of incorporating parasite studies to enhance our fundamental understanding of perturbations in host-parasite systems during translocation, in particular the site-level drivers of parasite dynamics.

Increased Trypanosoma spp. richness and prevalence of haemoparasite co-infection following translocation.

BACKGROUND: Understanding how fauna translocation and antiparasitic drug treatment impact parasite community structure within a host is vital for optimising translocation outcomes. Trypanosoma spp. and piroplasms (Babesia and Theileria spp.) are known to infect Australian marsupials, including the woylie (Bettongia penicillata). However relatively little is known about these haemoparasites, or how they respond to management practices such as translocation. We monitored haemoparasites infecting woylies for up to 12 months during two fauna translocations to supplement existing woylie populations in three different sites (Dryandra, Walcott and Warrup East) within south-western Australia between 2014 and 2016, with the aim of investigating (i) how haemoparasite prevalence, Trypanosoma spp. richness and Trypanosoma spp. community composition varied over time and between different sites following translocation; and (ii) whether ivermectin treatment indirectly impacts haemoparasite prevalence. Using molecular methods, 1211 blood samples were screened for the presence of trypanosomes, and a subset of these samples (n = 264) were also tested for piroplasms. RESULTS: Trypanosomes and piroplasms were identified in 55% and 94% of blood samples, respectively. We identified five Trypanosoma species, two Theileria species, a single species of Babesia and a novel Bodo species. Trypanosoma spp. richness and the prevalence of haemoparasite co-infection increased after translocation. Prior to translocation, Trypanosoma spp. community composition differed significantly between translocated and resident woylies within Walcott and Warrup East, but not Dryandra. Six months later, there was a significant difference between translocated and resident woylies within Dryandra, but not Walcott or Warrup East. The response of haemoparasites to translocation was highly site-specific, with predominant changes to the haemoparasite community in translocated woylies occurring within the first few months following translocation. Ivermectin treatment had no significant effect on haemoparasite prevalence. CONCLUSIONS: This study contributes to our understanding of haemoparasite dynamics in woylies following translocation. The highly site-specific and rapid response of haemoparasites to translocation highlights the need to better understand what drives these effects. Given that haemoparasite prevalence and composition of translocated and resident animals changed significantly following translocation, we propose that parasite monitoring should form an essential component of translocation protocols, and such protocols should endeavour to monitor translocated hosts and cohabiting species.

Debilitating disease in a polyparasitised woylie (Bettongia penicillata): A diagnostic investigation.

During monitoring of critically endangered woylie (Bettongia penicillata) populations within the south-west of Western Australia, an adult female woylie was euthanased after being found in extremely poor body condition with diffuse alopecia, debilitating skin lesions and severe ectoparasite infestation. Trypanosoma copemani G2 and Sarcocystis sp. were detected molecularly within tissue samples collected post-mortem. Potorostrongylus woyliei and Paraustrostrongylus sp. nematodes were present within the stomach and small intestine, respectively. Blood collected ante-mortem revealed the presence of moderate hypomagnesaemia, mild hypokalaemia, mild hyperglobulinaemia and mild hypoalbuminaemia. Diffuse megakaryocytic hypoplasia was evident within the bone marrow. We propose various hypotheses that may explain the presence of severe ectoparasite infection, skin disease and poor body condition in this woylie. Given the potential deleterious effects of parasite infection, the importance of monitoring parasites cannot be over-emphasised.

Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia.

In Western Australia a number of indigenous Trypanosoma spp. infect susceptible native marsupials, such as the woylie (Bettongia penicillata), brushtail possum (Trichosurus vulpecula), and chuditch (Dasyurus geoffroii). Two genotypes of Trypanosoma copemani (identified as G1 and G2) have been found in the woylie, and G2 has been implicated in the decline of this host species, making its presence of particular interest. Here we used targeted amplicon next generation sequencing (NGS) of the Trypanosoma 18S rDNA loci on 70 Trypanosoma-positive marsupial blood samples, to identify T. copemani genotypes and multiple Trypanosoma infections (polyparasitism) in woylies and cohabiting species in Western Australia. Polyparasitism with Trypanosoma spp. was found in 50% of the wildlife sampled, and within species diversity was high, with 85 zero-radius operational taxonomic units (ZOTUs) identified in nine putative parasite species. Trypanosoma copemani was assigned 17 ZOTUs and was identified in 80% of samples. The most abundant ZOTU isolated (63%) differed slightly from the published genotype of G1, and G2 was the second most abundant ZOTU (14%). Trypanosome diversity was significantly greater in woylies than in brushtail possums, and parasite community composition also differed significantly between these host species. One novel Trypanosoma spp. genotype (Trypanosoma sp. ANU2) was found in 20% of samples. A species of Crithidia was detected in a woylie, and two avian trypanosomes (Trypanosoma avium and Trypanosoma sp. AAT) were identified in woylies for the first time.

Trypanosome co-infections increase in a declining marsupial population.

Understanding the impacts of parasites on wildlife is growing in importance as diseases pose a threat to wildlife populations. Woylie (syn. brush-tailed bettong, Bettongia penicillata) populations have undergone enigmatic declines in south-western Western Australia over the past decade. Trypanosomes have been suggested as a possible factor contributing towards these declines because of their high prevalence in the declining population. We asked whether temporal patterns of infection with Trypanosoma spp. were associated with the decline patterns of the host, or if other factors (host sex, body condition, co-infection or rainfall) were more influential in predicting infection patterns. Species-specific nested PCRs were used to detect the two most common trypanosomes (T. copemani and T. vegrandis) from 444 woylie blood samples collected between 2006 and 2012. Time relative to the decline (year) and an interaction with co-infection by the other trypanosome best explained patterns of infection for both trypanosomes. The prevalence of single species infections for both T. copemani and T. vegrandis was lower after the population crash, however, the occurrence of co-infections increased after the crash compared to before the crash. Our results suggest an interaction between the two parasites with the decline of their host, leading to a higher level of co-infection after the decline. We discuss the possible mechanisms that may have led to a higher level of co-infection after the population crash, and highlight the importance of considering co-infection when investigating the role of parasites in species declines.

The effect of reinfection and mixed Trypanosoma cruzi infections on disease progression in mice.

The progression of Chagas disease (CD) varies significantly from host to host and is affected by multiple factors. In particular, mixed strain infections and reinfections have the potential to exacerbate disease progression subsequently affecting clinical management of patients with CD. Consequently, an associated reduction in therapeutic intervention and poor prognosis may occur due to this exacerbated disease state. This study investigated the effects of mixed strain infections and reinfection with Trypanosoma cruzi in mice, using two isolates from different discrete typing units, TcI (C8 clone 1) and TcIV (10R26). There were no significant differences in mortality rate, body weight or body condition among mice infected with either C8 clone 1, 10R26, or a mixture of both isolates. However, the parasite was found in a significantly greater number of host organs in mice infected with a mixture of isolates, and the histopathological response to infection was significantly greater in mice infected with C8 clone 1 alone, and C8 clone 1+10R26 mixed infections than in mice infected with 10R26 alone. To investigate the effects of reinfection, mice received either a double exposure to C8 clone 1; a double exposure to 10R26; exposure to C8 clone 1 followed by 10R26; or exposure to 10R26 followed by C8 clone 1. Compared to single infection groups, mortality was significantly increased, while survival time, body weight and body condition were all significantly decreased across all reinfection groups, with no significant differences among these groups. The mortality rate over all reinfection groups was 63.6%, compared to 0% in single infection groups, however there was no evidence of a greater histopathological response to infection. These results suggest firstly, that the C8 clone 1 isolate is more virulent than the 10R26 isolate, and secondly, that a more disseminated infection may occur with a mixture of isolates than with single isolates, although there is no evidence that mixed infections have a greater pathological effect. By contrast, reinfections do have major effects on host survivability and thus disease outcome. This confirms previous research demonstrating spontaneous deaths following reinfection, a phenomenon that to our knowledge has only been reported once before.

Evaluating Stress Physiology and Parasite Infection Parameters in the Translocation of Critically Endangered Woylies (Bettongia penicillata).

Translocation can be stressful for wildlife. Stress may be important in fauna translocation because it has been suggested that it can exacerbate the impact of infectious disease on translocated wildlife. However, few studies explore this hypothesis by measuring stress physiology and infection indices in parallel during wildlife translocations. We analysed faecal cortisol metabolite (FCM) concentration and endoparasite parameters (nematodes, coccidians and haemoparasites) in a critically endangered marsupial, the woylie (Bettongia penicillata), 1-3 months prior to translocation, at translocation, and 6 months later. FCM for both translocated and resident woylies was significantly higher after translocation compared to before or at translocation. In addition, body condition decreased with increasing FCM after translocation. These patterns in host condition and physiology may be indicative of translocation stress or stress associated with factors independent of the translocation. Parasite factors also influenced FCM in translocated woylies. When haemoparasites were detected, there was a significant negative relationship between strongyle egg count and FCM. This may reflect the influence of glucocorticoids on the immune response to micro- and macro-parasites. Our results indicate that host physiology and infection patterns can change significantly during translocation, but further investigation is required to determine how these patterns influence translocation success.

In vitro drug susceptibility of two strains of the wildlife trypanosome, Trypanosoma copemani: A comparison with Trypanosoma cruzi.

Trypanosomes are blood protozoan parasites that are capable of producing illness in the vertebrate host. Within Australia, several native Trypanosoma species have been described infecting wildlife. However, only Trypanosoma copemani has been associated with pathological lesions in wildlife hosts and more recently has been associated with the drastic decline of the critically endangered woylie (Bettongia penicillata). The impact that some trypanosomes have on the health of the vertebrate host has led to the development of numerous drug compounds that could inhibit the growth or kill the parasite. This study investigated and compared the in vitro susceptibility of two strains of T. copemani (G1 and G2) and one strain of Trypanosoma cruzi (10R26) against drugs that are known to show trypanocidal activity (benznidazole, posaconazole, miltefosine and melarsoprol) and against four lead compounds, two fenarimols and two pyridine derivatives (EPL-BS1937, EPL-BS2391, EPL-BS0967, and EPL-BS1246), that have been developed primarily against T.cruzi. The in vitro cytotoxicity of all drugs against L6 rat myoblast cells was also assessed. Results showed that both strains of T. copemani were more susceptible to all drugs and lead compounds than T. cruzi, with all IC50 values in the low and sub-µM range for both species. Melarsoprol and miltefosine exhibited the highest drug activity against both T. copemani and T. cruzi, but they also showed the highest toxicity in L6 cells. Interestingly, both fenarimol and pyridine derivative compounds were more active against T. copemani and T. cruzi than the reference drugs benznidazole and posaconazole. T. copemani strains exhibited differences in susceptibility to all drugs demonstrating once again considerable differences in their biological behaviour.

Host stress physiology and Trypanosoma haemoparasite infection influence innate immunity in the woylie (Bettongia penicillata).

Understanding immune function is critical to conserving wildlife in view of infectious disease threats, particularly in threatened species vulnerable to stress, immunocompromise and infection. However, few studies examine stress, immune function and infection in wildlife. We used a flow cytometry protocol developed for human infants to assess phagocytosis, a key component of innate immunity, in a critically endangered marsupial, the woylie (Bettongia penicillata). The effects of stress physiology and Trypanosoma infection on phagocytosis were investigated. Blood and faecal samples were collected from woylies in a captive facility over three months. Trypanosoma status was determined using PCR. Faecal cortisol metabolites (FCM) were quantified by enzyme-immunoassay. Mean phagocytosis measured was >90%. An interaction between sex and FCM influenced the percentage of phagocytosing leukocytes, possibly reflecting the influence of sex hormones and glucocorticoids. An interaction between Trypanosoma status and FCM influenced phagocytosis index, suggesting that stress physiology and infection status influence innate immunity.

Selection and optimization of hits from a high-throughput phenotypic screen against Trypanosoma cruzi.

BACKGROUND: Inhibitors of Trypanosoma cruzi with novel mechanisms of action are urgently required to diversify the current clinical and preclinical pipelines. Increasing the number and diversity of hits available for assessment at the beginning of the discovery process will help to achieve this aim. RESULTS: We report the evaluation of multiple hits generated from a high-throughput screen to identify inhibitors of T. cruzi and from these studies the discovery of two novel series currently in lead optimization. Lead compounds from these series potently and selectively inhibit growth of T. cruzi in vitro and the most advanced compound is orally active in a subchronic mouse model of T. cruzi infection. CONCLUSION: High-throughput screening of novel compound collections has an important role to play in diversifying the trypanosomatid drug discovery portfolio. A new T. cruzi inhibitor series with good drug-like properties and promising in vivo efficacy has been identified through this process.

A virus-free poly-promoter vector induces pluripotency in quiescent bovine cells under chemically defined conditions of dual kinase inhibition.

Authentic induced pluripotent stem cells (iPSCs), capable of giving rise to all cell types of an adult animal, are currently only available in mouse. Here, we report the first generation of bovine iPSC-like cells following transfection with a novel virus-free poly-promoter vector. This vector contains the bovine cDNAs for OCT4, SOX2, KLF4 and c-MYC, each controlled by its own independent promoter. Bovine fibroblasts were cultured without feeders in a chemically defined medium containing leukaemia inhibitory factor (LIF) and inhibitors of MEK1/2 and glycogen synthase kinase-3 signaling ('2i'). Non-invasive real-time kinetic profiling revealed a different response of bovine vs human and mouse cells to culture in 2i/LIF. In bovine, 2i was necessary and sufficient to induce the appearance of tightly packed alkaline phosphatase-positive iPSC-like colonies. These colonies formed in the absence of DNA synthesis and did not expand after passaging. Following transfection, non-proliferative primary colonies expressed discriminatory markers of pluripotency, including endogenous iPSC factors, CDH1, DPPA3, NANOG, SOCS3, ZFP42, telomerase activity, Tra-1-60/81 and SSEA-3/4, but not SSEA-1. This indicates that they had initiated a self-sustaining pluripotency programme. Bovine iPSC-like cells maintained a normal karyotype and differentiated into derivatives of all three germ layers in vitro and in teratomas. Our study demonstrates that conversion into induced pluripotency can occur in quiescent cells, following a previously undescribed route of direct cell reprogramming. This identifies a major species-specific barrier for generating iPSCs and provides a chemically defined screening platform for factors that induce proliferation and maintain pluripotency of embryo-derived pluripotent stem cells in livestock.

A comprehensive expression analysis of the Arabidopsis proline-rich extensin-like receptor kinase gene family using bioinformatic and experimental approaches.

The Arabidopsis proline-rich extensin-like receptor kinase (PERK) family consists of 15 predicted receptor kinases. A comprehensive expression analysis was undertaken to identify overlapping and unique expression patterns within this family relative to their phylogeny. Three different approaches were used to study AtPERK gene family expression, and included analyses of the EST, MPSS and NASCArrays databases as well as experimental RNA blot analyses. Some of the AtPERK members were identified as tissue-specific genes while others were more broadly expressed. While in some cases there was a good association between these different expression patterns and the position of the AtPERK members in the kinase phylogeny, in other cases divergence of expression patterns was seen. The PERK expression data identified by the bioinformatics and experimental approaches were found generally to show similar trends and supported the use of data from large-scale expression studies for obtaining preliminary expression data. Thus, the bioinformatics survey for ESTs and microarrays is a powerful comprehensive approach for obtaining a genome-wide view of genes in a multigene family.