Long-term field studies were conducted on two government managed small ruminant research farms, located in different geo-climatic regions and approximately 300 km separate from each other, on Peninsula Malaysia. The Infoternak trial (48 weeks) and the Chalok trial (43 weeks) each compared nematode parasite control in separately managed groups of young sheep, either short-term rotationally grazed around a suite of 10 paddocks in addition to receiving a daily supplement of Duddingtonia flagrans spores (Fungus Group); or similar groups of sheep being rotationally grazed alone (Control Group). The prevailing weather conditions at Infoternak farm were of below average rainfall conditions for the most of the trial. As a consequence, only very low worm infections (almost exclusively Haemonchus contortus) were acquired by the 17 sets of tracer lambs that grazed sequentially with the experimental lambs. However on all except 2 occasions in the early part of the trial, the mean tracer worm burdens were significantly lower (P < 0.05) and the experimental lambs grew significantly better (P = 0.054) in the Fungus Group. Rainfall at Chalok farm during the course of the trial was also below average. As a consequence infectivity of pastures was assumed to be relatively low based on faecal egg counts (epg) of the experimental sheep, which following an anthelmintic treatment prior to allocation, remained very low in both treatment groups. Faecal egg counts of undosed replacement lambs in the latter half of the Chalok study, showed a progressive increase in the Control Group to levels exceeding 3000 epg, whereas the Fungus Group remained static at approximately 500 epg. These results show that the deployment of the nematophagous fungus, D. flagrans, can improve the level of parasite control of sheep in the tropics above that which can be achieved by the short-term rotational grazing strategy alone.
Control of nematode parasites of small ruminants in a wet, tropical environment using the nematophagous fungus, Duddingtonia flagrans, was assessed in this study. Two methods of fungal delivery were tested, namely as a daily feed supplement, or incorporated into feed blocks. Initially, pen trials were conducted with individually penned groups of sheep and goats at dose rates of 125,000 spores and 250,000 spores/kg live weight per day. At the lower dose rate this reduction was between 80 and 90% compared with the pre-treatment levels. At the higher dose rate, there was virtually complete suppression (>99% reduction) of larval recovery. Trials using the fungal feed blocks, showed that when animals were individually penned, they consumed only small amounts of the block (particularly goats), hence little effect on larval recovery in faecal cultures was observed. Grouping animals according to species and dose rate induced satisfactory block consumption and subsequent high levels of larval reduction in faecal cultures. These larval reductions were mirrored by the presence of fungus in faecal cultures. This work was followed by a small paddock trial, whereby three groups of sheep were fed either a feed supplement without fungal spores, supplement with spores, or offered fungal blocks. The dose rate of spores in the latter two groups was 500,000 spores/kg live weight per day. Egg counts were significantly reduced in the two fungal groups, compared with the control group and the latter required two salvage anthelmintic treatments to prevent mortality due to haemonchosis. Pasture larval numbers on the two fungal group plots were also much lower than on the control plot.
A combination of genetic and functional approaches has identified three independent breast cancer risk loci at 2q35. A recent fine-scale mapping analysis to refine these associations resulted in 1 (signal 1), 5 (signal 2), and 42 (signal 3) credible causal variants at these loci. We used publicly available in silico DNase I and ChIP-seq data with in vitro reporter gene and CRISPR assays to annotate signals 2 and 3. We identified putative regulatory elements that enhanced cell-type-specific transcription from the IGFBP5 promoter at both signals (30- to 40-fold increased expression by the putative regulatory element at signal 2, 2- to 3-fold by the putative regulatory element at signal 3). We further identified one of the five credible causal variants at signal 2, a 1.4 kb deletion (esv3594306), as the likely causal variant; the deletion allele of this variant was associated with an average additional increase in IGFBP5 expression of 1.3-fold (MCF-7) and 2.2-fold (T-47D). We propose a model in which the deletion allele of esv3594306 juxtaposes two transcription factor binding regions (annotated by estrogen receptor alpha ChIP-seq peaks) to generate a single extended regulatory element. This regulatory element increases cell-type-specific expression of the tumor suppressor gene IGFBP5 and, thereby, reduces risk of estrogen receptor-positive breast cancer (odds ratio = 0.77, 95% CI 0.74-0.81, p = 3.1 × 10-31).