Spatial transcriptomics reveals alterations in perivascular macrophage lipid metabolism in the onset of Wooden Breast myopathy in broiler chickens.

This study aims to use spatial transcriptomics to characterize the cell-type-specific expression profile associated with the microscopic features observed in Wooden Breast myopathy. 1 cm3 muscle sample was dissected from the cranial part of the right pectoralis major muscle from three randomly sampled broiler chickens at 23 days post-hatch and processed with Visium Spatial Gene Expression kits (10X Genomics), followed by high-resolution imaging and sequencing on the Illumina Nextseq 2000 system. WB classification was based on histopathologic features identified. Sequence reads were aligned to the chicken reference genome (Galgal6) and mapped to histological images. Unsupervised K-means clustering and Seurat integrative analysis differentiated histologic features and their specific gene expression pattern, including lipid laden macrophages (LLM), unaffected myofibers, myositis and vasculature. In particular, LLM exhibited reprogramming of lipid metabolism with up-regulated lipid transporters and genes in peroxisome proliferator-activated receptors pathway, possibly through P. Moreover, overexpression of fatty acid binding protein 5 could enhance fatty acid uptake in adjacent veins. In myositis regions, increased expression of cathepsins may play a role in muscle homeostasis and repair by mediating lysosomal activity and apoptosis. A better knowledge of different cell-type interactions at early stages of WB is essential in developing a comprehensive understanding.

Effects of heat stress and a low energy diet on blood parameters and liver hsp70 and iNOS gene expressions in local chickens.

Two experiments were conducted to compare effects of heat stress and its combination with low dietary energy on blood indices, liver hsp70 and iNOS gene expressions in three Tanzanian local chicken ecotypes. In experiment one, five weeks old Kuchi (K), Ching'wekwe (C) and Morogoro medium (M) were randomly allocated to separate pens in a 3 × 2 factorial design in two adjacent rooms with controlled temperature. The study had three replicates consisting of 39 chickens per room, 13 per ecotype per pen making a total of 234 chickens. In one room, temperature was maintained at 26.5 ± 0.5 °C while in another it was maintained at 32±1 °C for 7 days and thereafter 37±1 °C for 10 days. A similar design was used in experiment two except that chickens were fed 55% less energy. In experiment one, serum corticosterone levels increased (p<0.05) in C and K. Gene expressions for hsp70 and iNOS were unchanged though hsp70 levels for K were higher (p<0.05). In experiment two, corticosterone levels were significantly elevated (p<0.05) in all ecotypes. Heterophil/lymphocyte ratios were markedly increased and changes in Hb and Hct at higher temperatures showed ecotype differences. Serum triglycerides were significantly reduced in all ecotypes. Hsp70 and iNOS levels were up-regulated in all ecotypes with levels in K higher (p<0.05) than in M. In both experiments, there were marked reductions in serum total protein. These results suggest that ecotype-based differences exist in local chickens' responses to heat stress and its combination with low energy diets. M and C demonstrated better tolerance than K when only heat stress was applied but a synergistic effect of heat stress and low dietary energy suggested M is more tolerant.

Effects of "Bioactive" amino acids leucine, glutamate, arginine and tryptophan on feed intake and mRNA expression of relative neuropeptides in broiler chicks.

Feed intake control is vital to ensuring optimal nutrition and achieving full potential for growth and development in poultry. The aim of the present study was to investigate the effects of L-leucine, L-glutamate, L-tryptophan and L-arginine on feed intake and the mRNA expression levels of hypothalamic Neuropeptide involved in feed intake regulation in broiler chicks. Leucine, glutamate, tryptophan or arginine was intra-cerebroventricularly (ICV) administrated to 4d-old broiler chicks respectively and the feed intake were recorded at various time points. Quantitative PCR was performed to determine the hypothalamic mRNA expression levels of Neuropeptide Y (NPY), agouti related protein (AgRP), pro-opiomelanocortin (POMC), melanocortin receptor 4 (MC4R) and corticotrophin releasing factor (CRF). Our results showed that ICV administration of L-leucine (0.15 or 1.5 µmol) significantly (P < 0.05) increased feed intake up to 2 h post-administration period and elevated both hypothalamic NPY and AgRP mRNA expression levels. In contrast, ICV administration of L-glutamate (1.6 µmol) significantly (P < 0.05) decreased feed intake 0.25, 0.5 and 2 h post-injection, and increased hypothalamic CRF and MC4R mRNA expression levels. Meanwhile, both L-tryptophan (10 or 100 µg) and L-arginine (20 or 200 µg) had no significant effect on feed intake. These findings suggested that L-leucine and L-glutamate could act within the hypothalamus to influence food intake, and that both orexigenic and anorexigenic Neuropeptide genes might contribute directly to these effects.