Unrecognized diversity in New Guinean crayfish species (Decapoda,Parastacidae): The evidence from molecular data

The phylogenetic relationships among imported ornamental crayfish belonging to the genus Cherax were inferred from a combined dataset of 3 mitochondrial genes (COI, 16S and 12S) and by comparison with available GenBank sequences of 14 Cherax species. Furthermore, the concordance of previously described species obtained from a wholesaler (Cherax boesemani, C. holthuisi and C. peknyi) with available GenBank sequences was verified based on COI with special respect to comparison with sequences assigned as Cherax species. Recently described species C. gherardiae, C. pulcher and C. subterigneus belong to the northern group of Cherax species. Comparison and analysis with other GenBank COI sequences show previously unreported diversity of New Guinean species, suggesting 5 putative new species. Surprisingly, species assigned to the subgenus Astaconephrops do not form a monophyletic clade; this subgenus should be reappraised relative to the purported typical morphological characteristic of the uncalcified patch on male chelae. Increasing importation of crayfish underscores the importance of accurate species identification. Use of basic molecular methods is a necessary requisite for documenting occurrence, abundance and population trends of target species. Consequently, it helps to support eventual conservation decision‐making by stakeholders.     

Pythium yorkensis sp. nov., a potential soybean pathogen from southeastern Pennsylvania,USA

Biodiversity surveys of natural as well as agricultural ecosystems commonly reveal novel isolates. A new species, Pythium yorkensis sp. nov., is reported here, isolated from field soil during a survey of oomycete diversity in symptomless soybean across southeastern Pennsylvania. Molecular data from both mitochondrial and nuclear loci support a unique phylogenetic position for the isolates collected, and morphological features distinguish this new species from close relatives in Pythium clade J. Pathogenicity assays suggest that this new species may be a potential agricultural pathogen, as isolates were mildly aggressive on soybean. This study highlights the continued need for culture‐based surveys in collaboration with high‐throughput sequence‐based approaches for environmental sampling.     

Interactions of Pratylenchus thornei and Fusarium oxysporum f. sp. ciceris on Chickpea

ABSTRACT Fusarium oxysporum f. sp. ciceris and the root-lesion nematode Pratylenchus thornei coinfect chickpeas in southern Spain. The influence of root infection by P. thornei on the reaction of Fusarium wilt-susceptible (CPS 1 and PV 61) and wilt-resistant (UC 27) chickpea cultivars to F. oxysporum f. sp. ciceris race 5 was investigated under controlled and field conditions. Severity of Fusarium wilt was not modified by coinfection of chickpeas by P. thornei and F. oxysporum f. sp. ciceris, in simultaneous or sequential inoculations with the pathogens. Root infection with five nematodes per cm(3) of soil and 5,000 chlamydospores per g of soil of the fungus resulted in significantly higher numbers of propagules of F. oxysporum f. sp. ciceris with the wilt-susceptible cultivar CPS 1, but not with the wilt-resistant one. However, infection with 10 nematodes per cm(3) of soil significantly increased root infection by F. oxysporum f. sp. ciceris in both cultivars, irrespective of fungal inoculum densities (250 to 2,000 chlamydospores per g of soil). Plant growth was significantly reduced by P. thornei infection on wilt-susceptible and wilt-resistant chickpeas in controlled and field conditions, except when shorter periods of incubation (45 days after inoculation) were used under controlled conditions. Severity of root necrosis was greater in wilt-susceptible and wilt-resistant cultivars when nematodes were present in the root, irrespective of length of incubation time (45 to 90 days), densities of nematodes (5 and 10 nematodes per cm(3) of soil), fungal inocula, and experimental conditions. Nematode reproduction on the wilt-susceptible cultivars, but not on the wilt-resistant one, was significantly increased by F. oxysporum f. sp. ciceris infections under controlled and field conditions.     

The α2A‐adrenoceptor subtype plays a key role in the analgesic and sedative effects of xylazine

Xylazine, the classical α₂‐adrenoceptor (α₂‐AR) agonist, is still used as an analgesic and sedative in veterinary medicine, despite its low potency and affinity for α₂‐ARs. Previous pharmacological studies suggested that the α_2A‐AR subtype plays a role in mediating the clinical effects of xylazine; however, these studies were hampered by the poor subtype‐selectivity of the antagonists used and a lack of knowledge of their bioavailability in vivo. Here, we attempted to elucidate the role of the α_2A‐AR subtype in mediating the clinical effects of xylazine by comparing the analgesic and sedative effects of this drug in wild‐type mice with those in α_2A‐AR functional knockout mice using the hot‐plate and open field tests, respectively. Hippocampal noradrenaline turnover in both mice was also measured to evaluate the contribution of α_2A‐AR subtype to the inhibitory effect of xylazine on presynaptic noradrenaline release. In wild‐type mice, xylazine (10 or 30 mg/kg) increased the hot‐plate latency. Furthermore, xylazine (3 or 10 mg/kg) inhibited the open field locomotor activity and decreased hippocampal noradrenaline turnover. By contrast, all of these effects were abolished in α_2A‐AR functional knockout mice. These results indicate that the α_2A‐AR subtype is mainly responsible for the clinical effects of xylazine.     

Lack of haplotype structuring for two candidate genes for trypanotolerance in cattle

Bovine trypanotolerance is a heritable trait associated to the ability of the individuals to control parasitaemia and anaemia. The INHBA (BTA4) and TICAM1 (BTA7) genes are strong candidates for trypanotolerance‐related traits. The coding sequence of both genes (3951 bp in total) were analysed in a panel including 79 Asian, African and European cattle (Bos taurus and B. indicus) to identify naturally occurring polymorphisms on both genes. In general, the genetic diversity was low. Nineteen of the 33 mutations identified were found just one time. Seventeen different haplotypes were defined for the TICAM1 gene, and 9 and 12 were defined for the exon 1 and the exon 2 of the INHBA gene, respectively. There was no clear separation between cattle groups. The most frequent haplotypes identified in West African taurine samples were also identified in other cattle groups including Asian zebu and European cattle. Phylogenetic trees and principal component analysis confirmed that divergence among the cattle groups analysed was poor, particularly for the INHBA sequences. The European cattle subset had the lowest values of haplotype diversity for both the exon1 (monomorphic) and the exon2 (0.077 ± 0.066) of the INHBA gene. Neutrality tests, in general, did not suggest that the analysed genes were under positive selection. The assessed scenario would be consistent with the identification of recent mutations in evolutionary terms.     

Competitive and allelopathic interference between soybean crop and annual wormwood (Artemisia annua L.) under field conditions

Annual wormwood interference on soybean crop growth and yield may result from competition and allelopathy, which are modulated by crop management. Allelochemicals released by annual wormwood (e.g. artemisinin) may affect the crop directly or indirectly through the effect on the nitrogen fixing symbiont, Bradyrhizobium japonicum. The objectives were (i) to quantify the crop response (i.e. biomass production, nodulation and yield) to weed interference and (ii) to determinate the relative change of competition and allelopathy interferences, when a sublethal dose of herbicide is applied. Two split plot field experiments with three replications were used. The experiment involved a factorial combination of five weed–crop density (soybean/annual wormwood, plants m^?2) levels: D1, pure soybean, 40/0 plants m^?2; D2, 40/2 plants m^?2; D3, 40/4 plants m^?2 and D4, 40/8 plants m^?2, and D5, pure annual wormwood, 0/8 plants m^?2, two activated carbon (allelopathy) levels: C?, with activated carbon (reduced allelopathy) and C+, non activated carbon applied (with allelopathy) and two herbicide levels: H?, untreated and H+, treated with a sub-lethal dose of glyphosate. Activated carbon to adsorb allelochemicals (with and without activated carbon) and glyphosate application (with and no herbicide) were assigned to sub-plots. Increasing weed density did not affect crop biomass at flowering, but changed nodule number and soybean yield with a different pattern depending on carbon and herbicide treatment. Relative crop yield decreased with increasing relative weed biomass. This decrease was particularly drastic when allelopathy was reduced by activated carbon and without herbicide application. The maximum yield losses of 33% in 2006 and 17% in 2007 were observed with the highest weed density (8 plants m^?2). In contrast, without carbon (high allelopathy level), soybean yield remained stable within the explored range of annual wormwood biomass, despite the fact that weed biomass at high densities (D4) was high enough to generate competition. The lack of response to increasing weed density could be related to the indirect effect of allelochemicals interacting with soil microorganisms (i.e. B. japonicum) that positively affected the nodulation (e.g. larger nodules in 2006 and increased nodules biomass due to higher number of roots in 2007 at high densities). With herbicide application, soybean yield of both carbon treatments remained stable when biomass of annual wormwood increased. This research provided strong evidence in support of the existence of positive effect of allelopathic and competitive interactions between annual wormwood and soybean crop under field conditions that may be overridden under herbicide application.     

Analysis of genomic mutation and immunohistochemistry of platelet‐derived growth factor receptors in canine vascular tumours

We examined whether mutation of the platelet‐derived growth factor receptor protein tyrosine kinase (PDGFR)‐α and PDGFR‐β genes contributes to their overexpression in canine vascular tumours. Genomic sequences of trans‐ or juxtamembrane regions of PDGFR‐α and PDGFR‐β were analysed with immunohistochemical staining and polymerase chain reaction‐direct sequencing using DNA from paraffin‐embedded neoplastic tissues of 27 hemangiosarcomas (HSAs) and 20 hemangiomas (HAs). Immunohistochemically, 75% of the HA cases were positive for PDGFR‐α and almost most of the HA cases were negative for PDGFR‐β. Of the HSA cases, 55.6% were negative for PDGFR‐α and 63% were strongly positive for PDGFR‐β. Among the HA cases, 1 missense mutation was detected in PDGFR‐α exon 18 and 1 in PDGFR‐β exon 17. Two HSA cases had missense mutations in exon 14 and 1 in exon 17 of PDGFR‐β. Thus, genomic mutation of trans‐ or juxtamembrane regions of PDGFRs was not the main mechanism driving the activation of receptors in HSA and HA.     

Effects of dietary Lactobacillus acidophilus and Bacillus subtilis on laying performance,egg quality,blood biochemistry and immune response of organic laying hens

The objective of this study was to evaluate the effects of two different probiotic micro‐organisms on the performance, egg quality and blood parameters of organically reared hens. A total of 900 16‐week‐old Hy‐Line layer hybrids were randomly assigned to three groups of 300 birds each. The control (CTR) group was fed a corn–soya bean cake‐based diet; the L group was fed the same diet supplemented with 0.1% Lactobacillus acidophilus, while the B group was fed the same diet supplemented with 0.05% Bacillus subtilis. Data were recorded at the beginning (weeks 5 and 6: T1) and at the end (weeks 19 and 20: T2) of the experiment, and no differences in hen performance were recorded between dietary groups or sampling times. All of the investigated clinical chemistry parameters, except GGT, were affected by diet (p < 0.05), with the best results recorded for the probiotic‐treated groups. The immune‐response values showed higher blood bactericidal activity in the B and L groups at T2 (p < 0.05) and a lower lysozime concentration in the B group at T1. Higher antibody production against Newcastle disease virus was observed in the L group compared to the CTR (p = 0.013). No differences in oxidative status were recorded, and no effects of diet on egg quality were observed. Among the physical egg characteristics, only the Roche scale colour was affected by diet (p < 0.05): the egg yolk was paler in the L group. The age of the hen was the most relevant factor affecting physical egg characteristics. The chemical parameters of the egg were almost unaffected by supplementation with probiotics except for the lipid content, which decreased with the L diet (p < 0.05). Both probiotic inclusions had beneficial effects on hen metabolism and welfare, and L. acidophilus induced the best immune response.