The myokine irisin is supposed to be cleaved from a transmembrane precursor, FNDC5 (fibronectin type III domain containing 5) and to mediate beneficial effects of exercise on human metabolism. However, evidence for irisin circulating in blood is largely based on commercial ELISA kits which are based on polyclonal antibodies (pAbs) not previously tested for cross-reacting serum proteins. We have analyzed four commercial pAbs by Western blotting, which revealed prominent cross-reactivity with non-specific proteins in human and animal sera. Using recombinant glycosylated and non-glycosylated irisin as positive controls, we found no immune-reactive bands of the expected size in any biological samples. A FNDC5 signature was identified at ~20 kDa by mass spectrometry in human serum but was not detected by the commercial pAbs tested. Our results call into question all previous data obtained with commercial ELISA kits for irisin and provide evidence against a physiological role for irisin in humans and other species.

Significance: Consequences of heat stress, particularly for the immune system and the intestinal health of mammals, are a topic of increasing global relevance due to rising temperatures and potential health impairments. Specific climate effects, however, are often difficult to discriminate from indirect consequences, e.g. reduced feed intake. Our study in dairy cattle, which are particularly sensitive to heat, identifies the infiltration of the small intestinal epithelium by a previously unobserved distinct cell population with macrophage-like phenotype in response to moderate heat stress. By using a pair-feeding design, we attributed these effects as direct consequences of heat stress via impaired intestinal barrier function. Therefore, an appropriate gut function is an important component in combating the negative consequences of heat stress.

The objective of this study was to investigate the growth- and breed-related changes of muscle fiber characteristics in cattle and their importance to meat quality. Four cattle breeds with different growth impetus and muscularity were reared and slaughtered under experimental conditions. German Angus as a beef type, Galloway as a hardy type, Holstein Friesian as a dairy type, and double-muscled Belgian Blue as an extreme type for muscle growth were used. Between 5 and 17 bulls of each breed were slaughtered at 0, 2, 4, 6, 12, 18, and 24 mo of age. Muscle fiber traits were determined and classified by computerized image analysis, and several measures of meat quality were also determined, including shear force value, meat color, and i.m. fat content. The postnatal growth of semitendinosus muscle in cattle was characterized by a nearly 10-fold increase of muscle fiber area from birth to 24 mo of age. In the first few months after birth, a transformation of type IIA fibers into IIB fibers was found, whereas type I fibers were nearly unaffected by age. The apparent total muscle fiber number of semitendinosus muscle did not increase during postnatal life. These results confirm that the fiber number is determined in embryonic development. Throughout the study, the double-muscled Belgian Blue (BBDM) bulls had almost twice the fiber number of the other breeds, emphasizing a more extensive hyperplasia of muscle fibers during embryonic development in BBDM compared with the other three breeds. The apparent number of type I fibers was, however, not affected by breed, which suggests that the additional fibers found in BBDM postnatally were type IIB and IIA fibers. We did not find significant differences in muscle fiber total number, muscle fiber type frequencies, or meat quality characteristics among breeds, with the exception of BBDM. Having pooled the four breeds, paler meat was related to a higher frequency of type IIB fibers, a lower area of type IIA and type I fiber, and a higher total muscle fiber number. These findings based on data of double muscling give us some hints for biological causes for the variation of meat quality. Further investigation, in particular within each breed, is necessary to identify the superior fiber traits for bovine meat production.