Ontogeny of pepsinogen and gastric proton pump expression in red porgy (Pagrus pagrus): Determination of stomach functionality

The appearance of functionally developed gastric glands is commonly considered as the transition from the larval to the juvenile stage in fish, since it means the switch from the less efficient alkaline digestion to a more efficient acid digestion characteristic of adult specimens. From that moment, fish are supposedly able to better assimilate nutrients from inert diets. Acid digestion takes place by the action of pepsin activity and hydrochloric acid, both secreted by the gastric glands of the stomach. Pepsinogen is the precursor of pepsin which is converted into active enzyme by the action of hydrochloric acid secreted by the proton pump. The goal of this work was to asses the ontogeny of pepsinogen and gastric proton pump expression along larval development of red porgy using RT-PCR and in situ hybridization techniques. Firstly, red porgy specific pepsinogen and proton pump partial sequences were isolated. Amplification products presented 615 and 591 bp and were identified as pepsinogen IIb and the -subunit of the proton pump (H⁺/K⁺-ATPase) by sequencing, respectively. Both sequences were aligned to several predicted pepsinogen and proton pump polypeptides from different vertebrate species showing elevated homologies with them, especially in the case of the proton pump, the identity of which was never less than 90%. Pepsinogen and proton pump expressions were detected from 30 days post-hatching (dph), increasing with development. Proton pump expression was localized in the gastric glands of red porgy larvae as revealed by in situ hybridization, showing increasing signal intensity along the digestive system development. Such results indicated that at 30 dph red porgy starts to acquire the adult digestive capacity and therefore inert diets should be better utilized from that time onwards.     

Serum pepsinogen levels to monitor gastrointestinal nematode infections in cattle revisited

The serum pepsinogen level is widely accepted as a useful parameter for monitoring gastrointestinal nematode infections in first-season grazing calves. However, several aspects, with possible implications for its practicality and the way to use the pepsinogen test, have received little attention to date. The objectives of this study were to evaluate (1) the reproducibility of a pepsinogen assay; (2) the required sample size for a reliable diagnosis and (3) the compliance to and effect of advice based on pepsinogen levels determined in year one on the nematode infection levels in the next generation of calves. Despite a high repeatability of the pepsinogen assay within the reference lab, the reproducibility between different labs was poor. There was more variation in pepsinogen levels between herds than between animals within a herd, suggesting that it is most useful to make a herd level diagnosis. Sample size calculation indicated that sampling seven animals sufficed to obtain a reliable indication of calf groups of up to 40 animals. Eighty-two herds were followed-up over two consecutive years and 39 (48%) had followed the advice that was formulated based on pepsinogen levels. Thirty-nine percent of the herds were advised to reduce the intensity of chemoprophylaxis. Samples from animals of those herds showed a higher pepsinogen level in the second year, but none of them had a mean pepsinogen level that is considered excessive. It is concluded that pepsinogen determination at housing from a limited number of animals can contribute to the design of the anthelmintic control strategy in the next year and a more targeted use of anthelmintics. However, more efforts are needed to harmonize the assay between veterinary diagnostic labs.     

The Contribution of Specific and Nonspecific Biomarkers in Diagnosis of Equine Gastric Ulcer Syndrome (EGUS) Under Field Condition

The aim of this study is to investigate the diagnostic efficiency of gastrin, pepsinogen, proinflammatory cytokines (TNF-α, IL-6), and oxidative stress biomarkers in horses with equine gastric ulcer syndrome (EGUS). Thirty horses diagnosed with gastroscopic EGUS and 15 clinically healthy horses were selected for this study. The serum levels of gastrin, pepsinogen showed nonsignificant changes in horses with EGUS when compared with healthy horses. The serum levels of TNF-α, IL-6 revealed a significant increase in horses with EGUS when compared with healthy ones. Oxidative stress is evident in horses with EGUS in comparison with healthy horses as detected by higher levels of malondialdehyde (MDA) and decreased serum levels of total antioxidant capacity (TAC), Superoxide dismutase (SOD), glutathione (GSH), and nitric oxide (NO). MDA and TNF-α showed better sensitivity and specificity than IL-6 in distinguishing horses with EGUS from control horses. Conclusively, examination of serum gastrin and pepsinogen levels had a limited value in diagnosis of EGUS in horses under investigation. Moreover, this study showed that oxidative stress is evident in horses with EGUS. Higher levels of TNF-α and IL-6 indicate their role in EGUS pathogenesis in horses. Finally, MDA, TNF-α, and IL-6 could be used as biological markers for preliminary screening of horses with EGUS. Gastroscopy still accredited as the “gold standard” for diagnosis EGUS.