Development of type I genetic markers from expressed sequence tags in highly polymorphic species

Expressed sequence tag (EST) databases provide a primary source of nuclear DNA sequences for genetic marker development in non-model organisms. To date, the process has been relatively inefficient for several reasons: - 1) priming site polymorphism in the template leads to inferior or erratic amplification; - 2) introns in the target amplicon are too large and/or numerous to allow effective amplification under standard screening conditions, and; - 3) at least occasionally, a PCR primer straddles an exon–intron junction and is unable to bind to genomic DNA template. The first is only a minor issue for species or strains with low heterozygosity but becomes a significant problem for species with high genomic variation, such as marine organisms with extremely large effective population sizes. Problems arising from unanticipated introns are unavoidable but are most pronounced in intron-rich species, such as vertebrates and lophotrochozoans. We present an approach to marker development in the Pacific oyster Crassostrea gigas, a highly polymorphic and intron-rich species, which minimizes these problems, and should be applicable to other non-model species for which EST databases are available. Placement of PCR primers in the 3′ end of coding sequence and 3′ UTR improved PCR success rate from 51% to 97%. Almost all (37 of 39) markers developed for the Pacific oyster were polymorphic in a small test panel of wild and domesticated oysters.

Projecting future expansion of invasive species: Comparing and improving methodologies for species distribution modeling

Modeling the distributions of species, especially of invasive species in non-native ranges, involves multiple challenges. Here, we developed some novel approaches to species distribution modeling aimed at reducing the influences of such challenges and improving the realism of projections. We estimated species-environment relationships with four modeling methods run with multiple scenarios of (1) sources of occurrences and geographically isolated background ranges for absences, (2) approaches to drawing background (absence) points, and (3) alternate sets of predictor variables. We further tested various quantitative metrics of model evaluation against biological insight. Model projections were very sensitive to the choice of training dataset. Model accuracy was much improved by using a global dataset for model training, rather than restricting data input to the species’ native range. AUC score was a poor metric for model evaluation and, if used alone, was not a useful criterion for assessing model performance. Projections away from the sampled space (i.e. into areas of potential future invasion) were very different depending on the modeling methods used, raising questions about the reliability of ensemble projections. Generalized linear models gave very unrealistic projections far away from the training region. Models that efficiently fit the dominant pattern, but exclude highly local patterns in the dataset and capture interactions as they appear in data (e.g. boosted regression trees), improved generalization of the models. Biological knowledge of the species and its distribution was important in refining choices about the best set of projections. A post-hoc test conducted on a new Partenium dataset from Nepal validated excellent predictive performance of our “best” model. We showed that vast stretches of currently uninvaded geographic areas on multiple continents harbor highly suitable habitats for Parthenium hysterophorus L. (Asteraceae; parthenium). However, discrepancies between model predictions and parthenium invasion in Australia indicate successful management for this globally significant weed. This article is protected by copyright. All rights reserved.

Placental infection with Ureaplasma species is associated with histologic chorioamnionitis and adverse outcomes in moderately preterm and late-preterm infants

Objective The human Ureaplasma species are the microbes most frequently isolated from placentae of women who deliver preterm. The role of Ureaplasma species has been investigated in pregnancies at <32 weeks of gestation, but currently no studies have determined the prevalence of ureaplasmas in moderately preterm and late-preterm (hereafter, “moderate/late preterm”) infants, the largest cohort of preterm infants. Methods Women delivering moderate/late preterm infants (n = 477) and their infants/placentae (n = 535) were recruited, and swab specimens of chorioamnion tissue, chorioamnion tissue specimens, and cord blood specimens were obtained at delivery. Swab and tissue specimens were cultured and analyzed by 16S ribosomal RNA polymerase chain reaction (PCR) for the presence of microorganisms, while cord blood specimens were analyzed for the presence of cytokines, chemokines, and growth factors. Results We detected microorganisms in 10.6% of 535 placentae (443 were delivered late preterm and 92 were delivered at term). Significantly, Ureaplasma species were the most prevalent microorganisms, and their presence alone was associated with histologically confirmed chorioamnionitis in moderate/late preterm and term placentae (P < .001). The presence of ureaplasmas in the chorioamnion was also associated with elevated levels of granulocyte colony-stimulating factor (P = .02). Conclusions These findings have important implications for infection and adverse pregnancy outcomes throughout gestation and should be of major consideration for obstetricians and neonatologists.

Projecting future expansion of invasive species: Comparing and improving methodologies for species distribution modeling

Modeling the distributions of species, especially of invasive species in non-native ranges, involves multiple challenges. Here, we developed some novel approaches to species distribution modeling aimed at reducing the influences of such challenges and improving the realism of projections. We estimated species-environment relationships with four modeling methods run with multiple scenarios of (1) sources of occurrences and geographically isolated background ranges for absences, (2) approaches to drawing background (absence) points, and (3) alternate sets of predictor variables. We further tested various quantitative metrics of model evaluation against biological insight. Model projections were very sensitive to the choice of training dataset. Model accuracy was much improved by using a global dataset for model training, rather than restricting data input to the species’ native range. AUC score was a poor metric for model evaluation and, if used alone, was not a useful criterion for assessing model performance. Projections away from the sampled space (i.e. into areas of potential future invasion) were very different depending on the modeling methods used, raising questions about the reliability of ensemble projections. Generalized linear models gave very unrealistic projections far away from the training region. Models that efficiently fit the dominant pattern, but exclude highly local patterns in the dataset and capture interactions as they appear in data (e.g. boosted regression trees), improved generalization of the models. Biological knowledge of the species and its distribution was important in refining choices about the best set of projections. A post-hoc test conducted on a new Partenium dataset from Nepal validated excellent predictive performance of our “best” model. We showed that vast stretches of currently uninvaded geographic areas on multiple continents harbor highly suitable habitats for Parthenium hysterophorus L. (Asteraceae; parthenium). However, discrepancies between model predictions and parthenium invasion in Australia indicate successful management for this globally significant weed. This article is protected by copyright. All rights reserved.

First report of Cotton leafroll dwarf virus in Thailand using a species-specific PCR validated with isolates from Brazil

Partial virus genome sequence with high nucleotide identity to Cotton leafroll dwarf virus (CLRDV) was identified from two cotton (Gossypium hirsutum) samples from Thailand displaying typical cotton leaf roll disease symptoms. We developed and validated a PCR assay for the detection of CLRDV isolates from Thailand and Brazil.

Fungus-feeding Thysanoptera: Phlaeothripinae of the Idiothrips genus-group in Australia, with nine new species

In a group of fungus-feeding Phlaeothripinae characterized by complex body sculpture, identification keys are provided to three genera and 15 species from Australia, including nine new species. In the genus Azaleothrips one new species is described, and one Asian species is newly recorded from Australia. The genus Stictothrips is recorded from Australia for the first time, with two new species. Within the genus Strepterothrips considerable structural diversity is recorded including three new species in which antennal segment III is greatly reduced and bears no sense cones. Some species in this genus exhibit the unusual condition of having several setae on the pelta, the first abdominal tergite. Problems in the production of generic diagnoses within the Phlaeothripinae are discussed.

Nomenclatural corrections to Australian species of Cricotopus (Wulp) (Diptera; Chironomidae)

Our attention has been drawn to lapsi and errors in a recent publication in this journal concerning Cricotopus Wulp (Diptera: Chironomidae) (Drayson et al., 2015).

2,6-Lutidine-N-oxide complexes of rare-earth bromides

2,6-Lutidine-N-oxide (LNO) complexes of rare-earth bromides of the composition $$MBr_3 .(LNO)_{4_{ - n} } .nH_2 O$$ wheren = l for M = La, Pr, Nd, Sm, Gd, Ho, Er; andn = 0 for M = Y have been prepared and characterised by analyses, conductance and infrared data. Infrared spectra of the complexes indicate that the coordination of ligand to the metal ion takes place through the oxygen of the ligand, and the water molecule in the complexes present is coordinated to the metal. A coordination number of seven has been suggested to all the rare-earth metal ions.

Topochemically controlled hydrogen reduction of scheelite-related rare-earth metal molybdates

Scheelite-related -Ln2Mo3O12(Ln = La, Pr, Nd, Sm, Gd, Tb, or Dy) oxides are reduced by hydrogen at 780–870 K yielding molybdenum (IV) oxides of formula Ln2Mo3O9. The latter crystallize in a tetragonal scheelite (ABO4) type structure where one third of the A sites and a quarter of the anion sites are vacant: Ln2/3(cat)1/3MoO3(an). The reaction Ln2Mo3O12+ 3H2 Ln2Mo3O9(an)3+ 3H2O may be regarded as topochemically controlled, since both the parent and the product phases have scheelite-related structures. Infrared spectra and electrical and magnetic properties of these metastable defect scheelite phases are reported.

Otolith morphology of four mackerel species (Scomberomorus spp.) in Australia: Species differentiation and prediction for fisheries monitoring and assessment

Four species of large mackerels (Scomberomorus spp.) co-occur in the waters off northern Australia and are important to fisheries in the region. State fisheries agencies monitor these species for fisheries assessment; however, data inaccuracies may exist due to difficulties with identification of these closely related species, particularly when specimens are incomplete from fish processing. This study examined the efficacy of using otolith morphometrics to differentiate and predict among the four mackerel species off northeastern Australia. Seven otolith measurements and five shape indices were recorded from 555 mackerel specimens. Multivariate modelling including linear discriminant analysis (LDA) and support vector machines, successfully differentiated among the four species based on otolith morphometrics. Cross validation determined a predictive accuracy of at least 96% for both models. An optimum predictive model for the four mackerel species was an LDA model that included fork length, feret length, feret width, perimeter, area, roundness, form factor and rectangularity as explanatory variables. This analysis may improve the accuracy of fisheries monitoring, the estimates based on this monitoring (i.e. mortality rate) and the overall management of mackerel species in Australia.

The upper respiratory tract is a natural reservoir of haemolytic Mannheimia species associated with ovine mastitis

Lamb suckling has been suggested to be an important way of infecting a ewe's udder with different bacteria, including Mannheimia haemolytica. To test the potential role of lambs in transferring Mannheimia species to the ewe’s udder, the restriction endonuclease cleavage patterns of isolates obtained from nasopharyngeal swabs were compared with those obtained from cases of mastitis. Sterile cotton swabs were used to collect nasopharyngeal samples from 50 ewes and 36 lambs from three flocks. M. haemolytica and Mannheimia glucosida as well as haemolytic Mannheimia ruminalis-like organisms were detected in the upper respiratory tract of lambs and ewes. Comparison of the restriction endonuclease cleavage patterns of the isolates suggested that the M. haemolytica isolates obtained from different milk samples from ewes with mastitis were more clonal than those obtained from the nasal swabs. However, some nasal isolates within both Mannheimia species had restriction endonuclease cleavage patterns identical to those obtained from milk samples from ewes with mastitis, indicating that lambs may have a role in transferring these organisms to the udder. More clonality was observed between the M. glucosida isolates than between M. haemolytica isolates.

Non-target species interaction with sodium fluoroacetate (1080) meat bait for controlling feral pigs (Sus scrofa)

Fresh meat baits containing sodium fluoroacetate (1080) are widely used for controlling feral pigs in Queensland, but there is a potential poisoning risk to non-target species. This study investigated the non-target species interactions with meat bait by comparing the time until first approach, investigation, sample and consumption, and whether dying bait green would reduce interactions. A trial assessing species interactions with undyed bait was completed at Culgoa Floodplain National Park, Queensland. Meat baits were monitored for 79 consecutive days with camera traps. Of 40 baits, 100% were approached, 35% investigated (moved) and 25% sampled, and 25% consumed. Monitors approached (P < 0.05) and investigated (P < 0.05) the bait more rapidly than pigs or birds, but the median time until first sampling was not significantly different (P > 0.05), and did not consume any entire bait. A trial was conducted at Whetstone State Forest, southern Queensland, with green-dyed and undyed baits monitored for eight consecutive days with cameras. Of 60 baits, 92% were approached and also investigated by one or more non-target species. Most (85%) were sampled and 57% were consumed, with monitors having slightly more interaction with undyed baits than with green-dyed baits. Mean time until first approach and sample differed significantly between species groups (P = 0.038 and 0.007 respectively) with birds approaching sooner (P < 0.05) and monitors sampling later (P < 0.05) than other (unknown) species (P > 0.05). Undyed bait was sampled earlier (mean 2.19 days) than green-dyed bait (2.7 days) (P = 0.003). Data from the two trials demonstrate that many non-target species regularly visit and sample baits. The use of green-dyed baits may help reduce non-target uptake, but testing is required to determine the effect on attractiveness to feral pigs. Further research is recommended to quantify the benefits of potential strategies to reduce the non-target uptake of meat baits to help improve the availability of bait to feral pigs.

Sirex woodwasp range expansion in Australia: performance and parasitism on two commercial pine species

Sirex woodwasp was detected in Queensland in 2009 and rapidly established in softwood plantations (Pinus radiata and P. taeda) in southern border regions. Biocontrol inoculations of Deladenus siricidicola began soon after, and adults were monitored to assess the success of the programme. Wasp size, sex ratios, emergence phenology and nematode parasitism rates were recorded, along with the assessment of wild-caught females. Patterns varied within and among seasons, but overall, P. taeda appeared to be a less suitable host than P. radiata, producing smaller adults, lower fat body content and fewer females. Sirex emerging from P. taeda also showed lower levels of nematode parasitism, possibly due to interactions with the more abundant blue-stain fungus in this host. Sirex adults generally emerged between November and March, with distinct peaks in January and March, separated by a marked drop in emergence in early February. Temperature provided the best correlate of seasonal emergence, with fortnights with higher mean minimum temperatures having higher numbers of Sirex emerging. This has implications for the anticipated northward spread of Sirex into sub-tropical coastal plantation regions. Following four seasons of inundative release of nematodes in Queensland, parasitism rates remain low and have resulted in only partial sterilization of infected females.

Responses of fruit flies (Tephritidae: Dacinae) to novel male attractants in north Queensland, Australia, and improved lures for some pest species

Male fruit fly attractants, cue-lure and methyl eugenol (ME), have been successfully used for the last 50 years in the monitoring and control of Dacini fruit flies (Bactrocera and Dacus species). However, over 50% of Dacini are non-responsive to either lure, including some pest species. A new lure, zingerone, has been found to weakly attract cue- and ME-responsive species in Malaysia. In Australia it attracted a weakly cue-responsive minor pest Bactrocera jarvisi (Tryon) and three non-responsive' species. Similar compounds were tested in Queensland and attracted cue- and ME-responsive species and two non-responsive' species. In this study, 14 novel compounds, including raspberry ketone formate (RKF) (Melolure) and zingerone, were field tested in comparison with cue-lure and ME at 17 sites in north Queensland. The most attractive novel lures were isoeugenol, methyl-isoeugenol, dihydroeugenol and zingerone. Several non-responsive' species responded to the new lures: Bactrocera halfordiae (Tryon), a species of some market access concern, was most attracted to isoeugenol; B.barringtoniae (Tryon), B.bidentata (May) and B.murrayi (Perkins) responded to isoeugenol, methyl-isoeugenol and dihydroeugenol; two new species of Dacus responded to zingerone. Bactrocera kraussi (Hardy), a cue-responsive minor pest in north Queensland, was significantly more attracted to isoeugenol than cue-lure. The cue-responsive D.absonifacies (May) and D.secamoneaeDrew were significantly more attracted to zingerone than cue-lure. Bactrocera yorkensisDrew & Hancock, a ME-responsive species was significantly more attracted to isoeugenol, methyl-isoeugenol and dihydroeugenol than ME. The preferential response to RKF or cue-lure was species specific. Six species were significantly more attracted to RKF, including the pests B.tryoni (Froggatt), B.frauenfeldi (Schiner) and minor pest B.bryoniae (Tryon); eight species were significantly more attracted to cue-lure including the pest B.neohumeralis (Hardy). These findings have significance in the search for optimal male lures for pest species elsewhere in the world.

Green and brown bridges between weeds and crops reveal novel Diaporthe species in Australia

Diaporthe (syn. Phomopsis) species are well-known saprobes, endophytes or pathogens on a range of plants. Several species have wide host ranges and multiple species may sometimes colonise the same host species. This study describes eight novel Diaporthe species isolated from live and/or dead tissue from the broad acre crops lupin, maize, mungbean, soybean and sunflower, and associated weed species in Queensland and New South Wales, as well as the environmental weed bitou bush (Chrysanthemoides monilifera subsp. rotundata) in eastern Australia. The new taxa are differentiated on the basis of morphology and DNA sequence analyses based on the nuclear ribosomal internal transcribed spacer region, and part of the translation elongation factor-1α and ß-tubulin genes. The possible agricultural significance of live weeds and crop residues ('green bridges') as well as dead weeds and crop residues ('brown bridges') in aiding survival of the newly described Diaporthe species is discussed.