Peptidases of Trichobilharzia regenti (Schistosomatidae) and its molluscan host Radix peregra S. Lat. (Lymnaeidae): construction and screening of cDNA library from intramolluscan stages of the parasite

Trichobilharzia regenti is a neurotropic bird schistosome,causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained.The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue.Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.     

Molecular Marker Analysis of Soybean Plant Introductions with Resistance to Phytophthora sojae

ABSTRACT Molecular analysis of sources of resistance to plant pathogens should expedite and confirm novel gene discovery and consequently the development of disease resistant cultivars. Recently, soybean plant introductions (PIs) were identified that contain putative novel Rps genes for resistance to Phytophthora sojae. The number of resistance genes that confer resistance to P. sojae isolates OH17 (1b,1d,2,3a,3b,3c,4,5,6,7) and OH25 (1a,1b,1c,1k,7) was then determined in several of the PIs. The objective of this study was to determine if the Rps genes present in these PIs were associated with eight described Rps loci that have been mapped on soybean molecular linkage groups F, G, J, and N. Nine F(2:3) soybean populations were genotyped with simple sequence repeat (SSR) markers linked to previously mapped Rps loci. The nine PI populations all had SSR markers associated (P < 0.01) with resistance to P. sojae isolate OH17 in the Rps1 region. Rps1c is a likely candidate in eight PIs but novel genes may also be possible, while novel genes may confer resistance in one PI to P. sojae isolate OHI7. Two or more Rps genes, including some that are potentially novel, confer resistance to P. sojae isolate OH25 in eight of the populations. However, based on the response to these two isolates, virulence already exists for at least some of the novel genes identified in this study.     

Genetic Analysis of Soybean Plant Introductions with Resistance to Phytophthora sojae

ABSTRACT Phytophthora sojae, which causes Phytophthora root and stem rot of soybean, is a serious disease worldwide and is managed primarily by deploying cultivars with resistance. Thirty-two soybean plant introductions (PIs), all but three of which were from South Korea, were proposed as new sources of single-gene resistance to P. sojae. The objective of this study was to characterize the inheritance of resistance to P. sojae in these PIs. Twenty-two soybean populations from crosses of these PIs and the susceptible cv. Williams were inoculated with P. sojae OH17 (vir 1b, 1d, 2, 3a, 3b, 3c, 4, 5, 6, 7), and OH25 (vir 1a, 1b, 1c, 1k, 7). These isolates were selected because they are virulent on soybeans with all known Rps genes and many Rps gene combinations. Thirteen of the twenty-two populations had consistent segregation responses following inoculations between the two generations. In two PIs, resistance was conferred by two genes to OH17 and three genes to OH25. Resistance to both isolates was conferred by a single gene in PI 398440 although the individual families were not resistant to the same isolates. The data suggest that six of the populations have three-Rps gene combinations as previously proposed, while another four may have either a novel Rps gene or a four-Rps gene combination. Based on this phenotypic analysis, novel and uncharacterized Rps genes may be present in this material. More importantly, these PIs may serve as sources of novel Rps genes that can be used to more effectively manage Phytophthora root and stem rot.     

Facultative and obligate diapause phenotypes in populations of the European spruce bark beetle Ips typographus

The bark beetle Ips typographus is the most destructive insect pest in Norway spruce-dominated forests. Its potential to establish multiple generations per year (multivoltinism) is one major trait that makes this beetle a severe pest. Ips typographus enters diapause to adjust its life cycle to seasonally changing environments. Diapause is characterized by developmental and reproductive arrest; it prolongs generation time and thus affects voltinism. In I. typographus a facultative, photoperiod-regulated diapause in the adult stage has been described. In addition, the presence of an obligate, photoperiod-independent, diapause has been hypothesized. The diapause phenotype has important implications for I. typographus voltinism, as populations with obligate diapausing individuals would be univoltine. To test for the presence of different I. typographus diapause phenotypes, we exposed Central and Northern European individuals to a set of photoperiodic treatments. We used two ovarian traits (egg number and vitellarium size) that are associated with gonad development, to infer reproductive arrest and thus diapause. We found a distinct effect of photoperiod on ovarian development, with variable responses in Central and Northern European beetles. We observed obligate diapausing (independent of photoperiod) individuals in Northern Europe, and both facultative (photoperiod-regulated) as well as obligate diapausing individuals in Central Europe. Our results show within-species variation for diapause induction, an adaptation to match life cycles with seasonally fluctuating environmental conditions. As the diapause phenotype affects the potential number of generations per season, our data are the basis for assessing the risk of outbreaks of this destructive bark beetle.

     

Integrated pest management of Tuta absoluta: practical implementations across different world regions

Journal of Pest Science - The South American tomato pinworm, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), has invaded most Afro-Eurasian countries and is threatening worldwide tomato...     

Disentangling the effect of topography and microtopography on near-ground growing-season frosts at the boreal-temperate forest ecotone (Québec,Canada)

New Forests - Permanent landscape attributes such as topography (elevation [m]) and microtopography (local variation in elevation [cm]) can increase the risk of cold air drainage down-slopes and in...     

Dietary Effects on Sperm Quality of Litopenaeus vannamei Broodstock

A 56‐d feeding trial was conducted to investigate the effect of diet on sperm quality of Pacific white shrimp Litopenaeus vannamei broodstock. Dietary treatments consisted of a combination of 75% dry maturation diet and 25% fresh‐frozen squid (dry‐weight basis). Supplemental nutrients of the maturation diet were selectively deleted and replaced with wheat starch to produce the following treatments: 1) 75% basal maturation diet plus 25% squid (control); 2) 75% maturation diet without supplemental vitamins plus 25% squid; 3) 75% maturation diet without supplemental cholesterol and phospholipids plus 25% squid; 4) 75% maturation diet without supplemental astaxanthin plus 25% squid; and 5) a fresh diet composed of 60% squid and 40% Maine bloodworms. Shrimp fed the control diet and the diet without supplemental astaxanthin had significantly higher mean (± SEM) change in sperm count (4.6 ± 3.2 million sperm cells and 2.9 ± 2.5 million sperm cells, respectively), with respect to baseline (8.7 ± 1.0, 6.4 ± 1.0, 9.0 ± 1.3, 6.6 ± 0.7, and 6.0 ± 0.8 million sperm cells for treatments 1, 2, 3, 4, and 5, respectively), than shrimp fed the diet without supplemental vitamins (‐1.7 ± 2.6 million sperm cells), but not significantly higher than those of shrimp receiving the diets without supplemental cholesterol‐phospholipids (1.2 ± 2.5 million sperm cells) and the fresh diet (1.3 ± 1.6 million sperm cells). Dietary deficiencies also were reflected in weight gain of shrimp fed the diet without supplemental vitamins (‐2.0 g) and the fresh diet (‐0.8 g). which were significantly lower than weight gain of shrimp fed the control diet (1.1 g) and the diet without supplemental cholesterol‐phospholipids (0.8 g). No significant differences were detected among treatments for percentage of abnormal sperm and survival data. Results demonstrated a significant effect of diet on reproductive quality of male L. vannamei and indicated that the typical combination of fresh‐food organisms used is not nutritionally optimal for male broodstock.     

Npro of Bungowannah virus exhibits the same antagonistic function in the IFN induction pathway than that of other classical pestiviruses

Bungowannah virus is the most divergent atypical pestivirus that had been detected up to now, and does not fit into any of the four approved species: Bovine viral diarrhea virus type 1 (BVDV-1) and type 2 (BVDV-2), Classical swine fever virus (CSFV) and Border disease virus (BDV). However, the presence of N^pro and E^rns coding regions, which are unique to pestiviruses, provides clear evidence of a pestivirus. Nevertheless, the amino acid identity of Bungowannah virus N^pro and BVDV-1 N^pro (strain CP7) is only 51.5%. By using a BVDV-1 backbone, a novel chimeric construct was generated, in which the genomic region encoding the non-structural protein N^pro was replaced by that of Bungowannah virus (CP7_N^pro-Bungo). In vitro studies of CP7_N^pro-Bungo revealed autonomous replication with the same efficacy as the BVDV backbone CP7 and infectious high-titer virus could be collected. In order to compare the ability of interferon (IFN) suppression, two reporter gene assays, specific for type-I IFN, were carried out. In virus-infected cells, no significant difference in blocking of IFN expression between the parental virus CP7, Bungowannah virus and the chimeric construct CP7_N^pro-Bungo could be detected. In contrast, an N^pro deletion mutant showed an impaired replication in bovine cells and a marked type-I IFN response.Taken together, our findings reveal the compatibility of non-structural protein N^pro of atypical Bungowannah virus with a BVDV type 1 backbone and its characteristic feature as an inhibitor of type-I IFN induction with an inhibitor-activity comparable to other pestiviruses.     

N‐Terminal Pro‐C‐Natriuretic Peptide and Cytokine Kinetics in Dogs with Endotoxemia

Background

Serum N‐terminal pro‐C‐natriuretic peptide (NT‐proCNP) concentration at hospital admission has sufficient sensitivity and specificity to differentiate naturally occurring sepsis from nonseptic systemic inflammatory response syndrome (SIRS). However, little is known about serum NT‐proCNP concentrations in dogs during the course of sepsis.

Objective

To determine serum NT‐proCNP and cytokine kinetics in dogs with endotoxemia, a model of canine sepsis.

Samples

Eighty canine serum samples.

Methods

Eight healthy adult Beagles were randomized to receive Escherichia coli lipopolysaccharide (LPS, 5 μg/kg) or placebo (0.9% NaCl) as a single IV dose in a randomized crossover study. Serum collected at 0, 1, 2, 4, and 24 hours was stored at −80°C for batch analysis. Serum NT‐proCNP was measured by ELISA and 13 cytokines and chemokines by multiplex magnetic bead‐based assay.

Results

Serum NT‐proCNP concentrations did not differ significantly between LPS‐ and placebo‐treated dogs at any time. When comparing serum cytokine concentrations, LPS‐treated dogs had higher interleukin‐6 (IL‐6), IL‐10, TNF‐α and KC‐like at 1, 2, and 4 hours; higher CCL2 at 1, 2, 4, and 24 hours; and higher IL‐8 and CXCL10 at 4 hours compared to placebo‐treated dogs. There were no differences in serum GM‐CSF, IFN‐γ, IL‐2, IL‐7, IL‐15 or IL‐18 between LPS‐ and placebo‐treated dogs.

Conclusions and Clinical Importance

Serum NT‐proCNP concentration does not change significantly in response to LPS administration in healthy dogs. Certain serum cytokine and chemokine concentrations are significantly increased within 1–4 hours after LPS administration and warrant further investigation as tools for the detection and management of sepsis in dogs.