Altered nociceptive, endocrine, and dorsal horn neuron responses in rats following a neonatal immune challenge

Summary The neonatal period is characterized by significant plasticity where the immune, endocrine, and nociceptive systems undergo fine-tuning and maturation. Painful experiences during this period can result in long-term alterations in the neurocircuitry underlying nociception, including increased sensitivity to mechanical or thermal stimuli. Less is known about the impact of neonatal exposure to mild inflammatory stimuli, such as lipopolysaccharide (LPS), on subsequent inflammatory pain responses. Here we examine the impact of neonatal LPS exposure on inflammatory pain sensitivity and HPA axis activity during the first three postnatal weeks. Wistar rats were injected with LPS (0.05 mg/kg IP, Salmonella enteritidis) or saline on postnatal days (PNDs) 3 and 5 and later subjected to the formalin test at PNDs 7, 13, and 22. One hour after formalin injection, blood was collected to assess corticosterone responses. Transverse spinal cord slices were also prepared for whole-cell patch clamp recording from lumbar superficial dorsal horn neurons (SDH). Brains were obtained at PND 22 and the hypothalamus was isolated to measure glucocorticoid (GR) and mineralocorticoid receptor (MR) transcript expression using qRT-PCR. Behavioural analyses indicate that at PND 7, no significant differences were observed between saline- or LPS-challenged rats. At PND 13, LPS-challenged rats exhibited enhanced licking (p < .01), and at PND 22, increased flinching in response to formalin injection (p < .05). LPS-challenged rats also displayed increased plasma corticosterone at PND 7 and PND 22 (p < .001) but not at PND 13 following formalin administration. Furthermore, at PND 22 neonatal LPS exposure induced decreased levels of GR mRNA and increased levels of MR mRNA in the hypothalamus. The intrinsic properties of SDH neurons were similar at PND 7 and PND 13. However, at PND 22, ipsilateral SDH neurons in LPS-challenged rats had a lower input resistance compared to their saline-challenged counterparts (p < .05). These data suggest neonatal LPS exposure produces developmentally regulated changes in formalin-induced behavioural responses, corticosterone levels, and dorsal horn neuron properties following noxious stimulation later in life. These findings highlight the importance of immune activation during the neonatal period in shaping pain sensitivity later in life. This programming involves both spinal cord neurons and the HPA axis.

Evaluation of growth and changes in body composition following neonatal diagnosis of cystic fibrosis

Early deficits in nutritional status that might require specific treatment and early response to nutritional therapy were studied longitudinally in 25 infants with cystic fibrosis (CF) diagnosed by neonatal screening, using anthropometric and research body composition methodology, and evaluation of pancreatic function. At the time of confirmed diagnosis (mean 5.4 weeks), body mass, length, total body fat (TBF), and total body potassium (TBK) were all significantly reduced. Following diagnosis and commencement of therapy there was a normalization of weight, length, and TBK by 6-12 months of age, indicating catch-up growth. But in some individuals the response was incomplete, and as a group, mean total body fat remained significantly lower than normal at 1 year of age. Seven of 25 (28%) were pancreatic sufficient at diagnosis, and all but one had evidence of declining pancreatic function requiring the institution of pancreatic enzyme therapy during the next 1-9 months. The median age of commencement of enzyme therapy was 10 weeks (range 5 weeks to 11 months). These longitudinal assessments emphasize the dynamic changes occurring in absorptive function, body composition, and nutritional status following neonatal diagnosis of cystic fibrosis and may reflect previously described abnormalities of energy metabolism in this age group. Abnormal body composition is evident in most CF infants following diagnosis by neonatal screening but pancreatic damage may still be evolving. We suggest that early active nutritional therapy and surveillance for changes in pancreatic function are warranted in CF infants diagnosed by neonatal screening.

Genetic analysis of resistance to root-lesion nematode (Pratylenchus thornei) in wheat

The inheritance of resistance to root-lesion nematode was investigated in five synthetic hexaploid wheat lines and two bread wheat lines using a half-diallel design of F1 and F2 crosses. The combining ability of resistance genes in the synthetic hexaploid wheat lines was compared with the performance of the bread wheat line 'GS50a', the source of resistance to Pratylenchus thornei used in Australian wheat breeding programmes. Replicated glasshouse trials identified P. thornei resistance as polygenic and additive in gene action. General combining ability (GCA) of the parents was more important than specific combining ability (SCA) effects in the inheritance of P. thornei resistance in both F1 and F2 populations. The synthetic hexaploid wheat line 'CPI133872' was identified as the best general combiner, however, all five synthetic hexaploid wheat lines possessed better GCA than 'GS50a'. The synthetic hexaploid wheat lines contain novel sources of P. thornei resistance that will provide alternative and more effective sources of resistance to be utilized in wheat breeding programmes

Neonatal screening for cystic fibrosis

Two groups of patients with cystic fibrosis were compared. The screened group, detected with an improved neonatal screening assay for immunoreactive trypsin, developed fewer chest infections requiring treatment and gained more weight than the unscreened group. Early diagnosis by screening seems to affect early morbidity.

Neonatal intestinal obstruction associated with oral calcium supplementation

A case report of a 920 g infant developing a small intestinal obstruction following therapy for congestive cardiac failure is presented. Although the causation was thought to be milk curd obstruction, subsequent analysis revealed high concentration of calcium and phosphate in the stools. The possible pathogenesis is discussed in relation to the inspissated milk syndrome.

Root-lesion nematodes (Pratylenchus thornei and P. neglectus): A review of recent progress in managing a significant pest of grain crops in northern Australia

Two species of root-lesion nematode (predominantly Pratylenchus thornei but also P. neglectus) are widespread pathogens of wheat and other crops in Australia's northern grain belt, a subtropical region with deep, fertile clay soils and a summer-dominant rainfall pattern. Losses in grain yield from P. thornei can be as high as 70% for intolerant wheat cultivars. This review focuses on research which has led to the development of effective integrated management programs for these nematodes. It highlights the importance of correct identification in managing Pratylenchus species, reviews the plant breeding work done in developing tolerant and resistant cultivars, outlines the methods used to screen for tolerance and resistance, and discusses how planned crop sequencing with tolerant and partially resistant wheat cultivars, together with crops such as sorghum, sunflower, millets and canaryseed, can be used to reduce nematode populations and limit crop damage. The declining levels of soil organic matter in cropped soils are also discussed with reference to their effect on soil health and biological suppression of root-lesion nematodes.

Identification of quantitative trait loci for resistance to two species of root-lesion nematode (Pratylenchus thornei and P. neglectus) in wheat

Pratylenchus thornei and P. neglectus are two species of root-lesion nematode that cause substantial yield losses in wheat. No commercially available wheat variety has resistance to both species. A doubled-haploid population developed from a cross between the synthetic hexaploid wheat line CPI133872 and the bread wheat Janz was used to locate and tag quantitative trait loci (QTLs) associated with resistance to both P. thornei and P. neglectus. Wheat plants were inoculated with both species of nematode in independent replicated glasshouse trials repeated over 2 years. Known locations of wheat microsatellite markers were used to construct a framework map. After an initial single-marker analysis to detect marker-trait linkages, chromosome regions associated with putative QTLs were targetted with microsatellite markers to increase map density in the chromosome regions of interest. In total, 148 wheat microsatellite markers and 21 amplified fragment length polymorphism markers were mapped. The codominant microsatellite marker Xbarc183 on the distal end of chromosome 6DS was allelic for resistance to both P. thornei and P. neglectus. The QTL were designated QRlnt.lrc-6D.1 and QRlnn.lrc-6D.1, for the 2 traits, respectively. The allele inherited from CPI133872 explained 22.0-24.2% of the phenotypic variation for P. thornei resistance, and the allele inherited from Janz accounted for 11.3-14.0% of the phenotypic variation for P. neglectus resistance. Composite interval mapping identified markers that flank a second major QTL on chromosome 6DL (QRlnt.lrc-6D.2) that explained 8.3-13.4% of the phenotypic variation for P. thornei resistance. An additional major QTL associated with P. neglectus resistance was detected on chromosome 4DS (QRlnn.lrc-4D.1) and explained a further 10.3-15.4% of the phenotypic variation. The identification and tagging of nematode resistance genes with molecular markers will allow appropriate allele combinations to be selected, which will aid the successful breeding of wheat with dual nematode resistance.

Resistance to the root-lesion nematode Pratylenchus thornei of Iranian landrace wheat.

Pratylenchus thornei is widespread throughout the wheat-growing regions in Australia and overseas and can cause yield losses of up to 70% in some intolerant cultivars. The most effective forms of management of P. thornei populations are crop rotation and plant breeding. There have been no wheat accessions identified as completely resistant to P. thornei, therefore breeding programs have used moderately resistant parents. The objective of the present research was to evaluate 274 Iranian landrace wheats for resistance to P. thornei and identify accessions with resistance superior to the current best resistance source (GS50a). Plants were grown in P. thornei inoculated soil under controlled conditions in a glasshouse pot experiment for 16 weeks. Ninety-two accessions found to be resistant or moderately so were retested in a second experiment. From combined analysis of these experiments, 34 accessions were identified as resistant with reproduction factors (final population per kg soil/initial inoculum rate per kg soil) <= 1. In total, 25 accessions were more resistant than GS50a, with AUS28470 significantly (P < 0.05) more resistant. The resistant Iranian landraces identified in the present study are a valuable untapped genetic pool offering improved levels of P. thornei resistance over current parents in Australian wheat-breeding programs.

Resistance to root-lesion nematodes (Pratylenchus thornei and P-neglectus) in synthetic hexaploid wheats and their durum and Aegilops tauschii parents.

Root-lesion nematodes (Pratylenchus thornei Sher and Allen and P. neglectus (Rensch) Filipijev and Schuurmans Stekhoven) cause substantial yield loss to wheat crops in the northern grain region of Australia. Resistance to P. thornei for use in wheat breeding programs was sought among synthetic hexaploid wheats (2n= 6x = 42, AABBDD) produced through hybridisations of Triticum turgidum L. subsp. durum (Desf.) Husn (2n= 4x = 28, AABB) with Aegilops tauschii Coss. (2n= 2x = 14, DD). Resistance was determined for the synthetic hexaploid wheats and their durum and Ae. tauschii parents from the numbers of nematodes in the roots of plants grown for 16 weeks in pots of pasteurised soil inoculated with P. thornei. Fifty-nine (32%) of 186 accessions of synthetic hexaploid wheats had lower numbers of nematodes than Gatcher Selection 50a (GS50a), a partially resistant bread wheat. Greater frequencies of partial resistance were present in the durum parents (72% of 39 lines having lower nematode numbers than GS50a) and in the Ae. tauschii parents (55% of 53 lines). The 59 synthetic hexaploids were re-tested in a second experiment along with their parents. In a third experiment, 11 resistant synthetic hexaploid wheats and their F-1 hybrids with Janz, a susceptible bread wheat, were tested and the F(1)s were found to give nematode counts intermediate between the respective two parents. Synthetic hexaploid wheats with higher levels of resistance resulted from hybridisations where both the durum and Ae. tauschii parents were partially resistant, rather than where only one parent was partially resistant. These results suggest that resistance to P. thornei in synthetic hexaploid wheats is polygenic, with resistances located both in the D genome from Ae. tauschii and in the A and/or B genomes from durum. Five synthetic hexaploid wheats were selected for further study on the basis of (1) a high level of resistance to P. thornei of the synthetic hexaploid wheats and of both their durum and Ae. tauschii parents, (2) being representative of both Australian and CIMMYT (International Maize and Wheat Improvement Centre) durums, and (3) being representative of the morphological subspecies and varieties of Ae. tauschii. These 5 synthetic hexaploid wheats were also shown to be resistant to P. neglectus, whereas GS50a and 2 P. thornei-resistant derivatives were quite susceptible. Results of P. thornei resistance of F(1)s and F(2)s from a half diallel of these 5 synthetic hexaploid wheats, GS50a, and Janz from another study indicate polygenic additive resistance and better general combining ability for the synthetic hexaploid wheats than for GS50a. Published molecular marker studies on a doubled haploid population between the synthetic hexaploid wheat with best general combining ability (CPI133872) and Janz have shown quantitative trait loci for resistance located in all 3 genomes. Synthetic hexaploid wheats offer a convenient way of introgressing new resistances to P. thornei and P. neglectus from both durum and Ae. tauschii into commercial bread wheats.

Resistance to the root-lesion nematode Pratylenchus thornei in wheat landraces and cultivars from the West Asia and North Africa (WANA) region.

Resistance to the root-lesion nematode Pratylenchus thornei was sought in wheat from the West Asia and North Africa (WANA) region in the Watkins Collection (148 bread and 139 durum wheat accessions) and the McIntosh Collection (59 bread and 43 durum wheat accessions). It was considered that landraces from this region, encompassing the centres of origin of wheat and where P. thornei also occurs, could be valuable sources of resistance for use in wheat breeding. Resistance was determined by number of P. thornei/kg soil after the growth of the plants in replicated glasshouse experiments. On average, durum accessions produced significantly lower numbers of P. thornei than bread wheat accessions in both the Watkins and McIntosh Collections. Selected accessions with low P. thornei numbers were re-tested and 13 bread wheat and 10 durum accessions were identified with nematode numbers not significantly different from GS50a, a partially resistant bread wheat line used as a reference standard. These resistant accessions, which originated in Iran, Iraq, Syria, Egypt, Sudan, Morocco, and Tunisia, represent a resource of resistance genes in the primary wheat gene pool, which could be used in Australian wheat breeding programs to reduce the economic loss from P. thornei.

QTL mapping of multiple foliar disease and root-lesion nematode resistances in wheat.

A genetic linkage map, based on a cross between the synthetic hexaploid CPI133872 and the bread wheat cultivar Janz, was established using 111 F1-derived doubled haploid lines. The population was phenotyped in multiple years and/or locations for seven disease resistance traits, namely, Septoria tritici blotch (Mycosphaeralla graminicola), yellow leaf spot also known as tan spot (Pyrenophora tritici-repentis), stripe rust (Puccinia striiformis f. sp. tritici), leaf rust (Puccinia triticina), stem rust (Puccinia graminis f. sp. tritici) and two species of root-lesion nematode (Pratylenchyus thornei and P. neglectus). The DH population was also scored for coleoptile colour and the presence of the seedling leaf rust resistance gene Lr24. Implementation of a multiple-QTL model identified a tightly linked cluster of foliar disease resistance QTL in chromosome 3DL. Major QTL each for resistance to Septoria tritici blotch and yellow leaf spot were contributed by the synthetic hexaploid parent CPI133872 and linked in repulsion with the coincident Lr24Sr24/ locus carried by parent Janz. This is the first report of linked QTL for Septoria tritici blotch and yellow leaf spot contributed by the same parent. Additional QTL for yellow leaf spot were detected in 5AS and 5BL. Consistent QTL for stripe rust resistance were identified in chromosomes 1BL, 4BL and 7DS, with the QTL in 7DS corresponding to the Yr18Lr34/ region. Three major QTL for P. thornei resistance (2BS, 6DS, 6DL) and two for P. neglectus resistance (2BS, 6DS) were detected. The recombinants combining resistance to Septoria tritici blotch, yellow leaf spot, rust diseases and root-lesion nematodes from parents CPI133872 and Janz constitute valuable germplasm for the transfer of multiple disease resistance into new wheat cultivars.

Inheritance of resistance to root-lesion nematode (Pratylenchus thornei) in wheat landraces and cultivars from the West Asia and North Africa (WANA) region.

The root-lesion nematode Pratylenchus thornei causes substantial loss to bread wheat production in the northern grain region of Australia and other parts of the world. West Asia and North Africa (WANA) wheat accessions with partial resistance to P. thornei were analysed for mode of inheritance in a half-diallel crossing design of F1 hybrids (10 parents) and F2 populations (7 parents). General combining ability was more important than specific combining ability as indicated by components of variance ratios of 0.93 and 0.95 in diallel ANOVA of the F1 and F2 generations, respectively. General combining ability values of the 'resistant' parents were predictive of the mean nematode numbers of their progeny in crosses with the susceptible Australian cv. Janz at the F1 (R populations showed relatively continuous distributions. Heritability was 0.68 for F2 populations in the half-diallel of resistant parents and 0.82-0.92 for 5 'resistant' parent/Janz doubled-haploid populations (narrow-sense heritability on a line mean basis). The results indicate polygenic inheritance of P. thornei resistance with a minimum of from 2 to 6 genes involved in individual F populations of 5 resistant parents crossed with Janz. Morocco 426 and Iraq 43 appear to be the best of the parents tested for breeding for resistance to P. thornei. None of the P. thornei-resistant WANA accessions was resistant to Pratylenchus neglectus.

Biological suppression of root-lesion nematodes in grain-growing soils

Root-lesion nematodes (RLNs) are found on 75% of grain farms in southern Queensland (QLD) and northern New South Wales (NSW) and are significant pests. This project confirmed that biological suppression of RLNs occurs in soils, examined what organisms are involved and how growers might enhance suppressiveness of soils. Field trials, and glasshouse and laboratory bioassays of soils from fields with contrasting management practices, showed suppressiveness is favoured with less tillage, more stubble and continuous intensive cropping, particularly in the top 15cm of soil. Through extensive surveys key organisms, Pasteuria bacteria, nematode-trapping fungi and predatory nematodes were isolated and identified as being present.