Hemp products are currently enjoying much commercial interest as potential anti-inflammatory preparations. However, evidence for efficacy is lacking for some clinical conditions and for crude extracts and mixtures of cannabinoids. Uterine inflammation (endometritis) is a major cause of subfertility in livestock. The aim was to assess whether a commercially important crude hemp extract (extracted with supercritical CO2) and a commercial CBG:CBD (cannabigerol:cannabidiol) cannabinoid mixture have anti-inflammatory properties, using an ex vivo bovine tissue culture model of endometritis. LC-MS2 (liquid chromatography-tandem mass spectrometry) analysis of the crude extract allowed identification of twelve cannabinoids (cannabidiolic acid (CBDA), cannabidiol (CBD), cannabidivarinic acid (CBDVA), cannabidivarin (CBDV), cannabielsoic acid A (CBEA-A), cannabielsoic acid B (CBEA-B), cannabichromene (CBC), cannabinoloic acid (CBNA), cannabinol (CBN), cannabiripsol (CBR), cannabicoumarononic acid (CBCONA), cannabicoumaronone (CBCON)) in the extract, along with isomers, an alkaloid and three polyunsaturated fatty acids. Fragmentation pathways, some including 6-jr-electron pericyclic processes, are proposed. Tissue explants, from uteri from heifers at stage I or IV of the oestrous cycle, were treated with control, LPS (lipopolysaccharide), crude hemp extract, or with LPS + crude hemp extract for 24 h. The CBG:CBD mixture was assessed similarly. Inflammatory responses were measured as concentrations of PGF2a, PGE2 and IL-6 in culture supernatants. Secretion of IL6 from tissue explants was abrogated by the hemp extract (50mg mL-1), compared to LPS alone, but it had no effect on secretion of PGF2a. The CBG:CBD mixture had no effect on any of the markers. The crude hemp extract was not cytotoxic towards MDBK bovine kidney cells whereas the CBG:CBD mixture (50 mg mL-1) caused cell death. The hemp extract warrants further investigation for anti-inflammatory effects using a broader range of inflammatory markers, whereas the CBG:CBD mixture is not recommended for further study in the context of bovine endometritis. (c) 2025 The Authors. Published by Elsevier B.V. on behalf of SAAB. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

The most common equine tapeworm, Anoplocephala perfoliata , has often been neglected amongst molecular investigations and has been faced with limited treatment options. However, the recent release of a transcriptome dataset has now provided opportunities for in-depth analysis of A. perfoliata protein expression. Here, global, and sub-proteomic approaches were utilized to provide a comprehensive characterization of the A. perfoliata soluble glutathione transferases (GST) (ApGST). Utilizing both bioinformatics and gel-based proteomics, GeLC and 2D-SDS PAGE, the A. perfoliata ‘ GST-ome’ was observed to be dominated with Mu class GST representatives. In addition, both Sigma and Omega class GSTs were identified, albeit to a lesser extent and absent from affinity chromatography approaches. Moreover, 51 ApGSTs were localized across somatic (47 GSTs), extracellular vesicles (EVs) (Whole: 1 GST, Surface: 2 GSTs) and EV depleted excretory secretory product (ESP) (9 GSTs) proteomes. In related helminths, GSTs have shown promise as novel anthelmintic or vaccine targets for improved helminth control. Thus, provides potential targets for understanding A. perfoliata novel infection mechanisms, host–parasite relationships and anthelmintic treatments.

Abstract Background Endometritis is an inflammatory condition of the uterus leading to complications with reproductive organs, miscarriage and infertility. The first-line treatment of endometritis in pigs are antibiotics, which do not always resolve inflammation and contribute to antibiotic resistance. There is an urgent need for the development of alternative treatments for endometritis. Here we propose and validate the first porcine tissue explant model for uterine inflammation and test naturally derived compounds for their effectiveness as anti-inflammatory therapies in porcine endometrial tissue. Methods Uterine endometrial explants were harvested from uteri of sows sent to slaughter. Tissue biopsies were collected at follicular and luteal phases of the oestrous cycle. Biopsies were washed and these tissue ‘explants’ were cultured. Explants were treated at 24 hr with oxytocin (to stimulate PGF2α production) or LPS (to mimic infection and induce inflammation. Explants were retained for RT-PCR analysis. Supernatants were harvested and concentrations of markers were measured by ELISA and Luminex. Results The data generated thus far show a dose dependent response of IL-6 secretion to LPS stimulation, which demonstrates the validity of our model. Inflammation can be stimulated in an in vitro setting to mimic porcine endometritis and we present data to show the change in cytokine levels in response to treatment with naturally derived compounds. Conclusions This data shows that inflammation can be stimulated in an in vitro setting to mimic porcine endometritis This model is a tool for testing of novel compounds and their potential anti-inflammatory effects on the porcine endometrium. Supported by an Innovate UK Grant (Project Reference: 98015) (https://gtr.ukri.org/projects?ref=98015)

Modelling plays an important role in assessing disease risks and the efficacy of preventative actions. However, the extent to which existing models meet the needs of different groups of dairy farmers around disease prevention is unclear. A questionnaire gathered information on disease prevention actions undertaken by organic and conventional dairy farmers in Spain and the UK, and on their information preferences and needs in relation to such actions. A systematic review of animal health modelling articles was undertaken to compare the expressed needs of dairy farmers for information on disease prevention, with the focus and outputs of existing models. Farmer groups differed in needs when planning disease prevention interventions. Most farmers sourced animal health information from veterinarians. Farmers preferred to use practical experience to judge the efficacy of change. To fulfil the expressed needs of dairy farmers, models need to address specific farming contexts and non-economic impacts of change.

Anoplocephala perfoliata is a neglected gastro-intestinal tapeworm, commonly infecting horses worldwide. Molecular investigation of A. perfoliata is hampered by a lack of tools to better understand the host–parasite interface. This interface is likely influenced by parasite derived immune modulators released in the secretome as free proteins or components of extracellular vesicles (EVs). Therefore, adult RNA was sequenced and de novo assembled to generate the first A. perfoliata transcriptome. In addition, excretory secretory products (ESP) from adult A. perfoliata were collected and EVs isolated using size exclusion chromatography, prior to proteomic analysis of the EVs, the EV surface and EV depleted ESP. Transcriptome analysis revealed 454 sequences homologous to known helminth immune modulators including two novel Sigma class GSTs, five α-HSP90s, and three α-enolases with isoforms of all three observed within the proteomic analysis of the secretome. Furthermore, secretome proteomics identified common helminth proteins across each sample with known EV markers, such as annexins and tetraspanins, observed in EV fractions. Importantly, 49 of the 454 putative immune modulators were identified across the secretome proteomics contained within and on the surface of EVs in addition to those identified in free ESP. This work provides the molecular tools for A. perfoliata to reveal key players in the host–parasite interaction within the horse host.

Preimplantation factor (PIF) is an embryo derived peptide which exerts an immune modulatory effect on human endometrium, promoting immune tolerance to the embryo whilst maintaining the immune response to invading pathogens. While bovine embryos secrete PIF, the effect on the bovine endometrium is unknown. Maternal recognition of pregnancy is driven by an embryo-maternal cross talk, however the process differs between humans and cattle. As many embryos are lost during the early part of pregnancy in cattle, a greater knowledge of factors affecting the embryo-maternal crosstalk, such as PIF, is needed to improve fertility. Therefore, for the first time, we demonstrate the effect of synthetic PIF (sPIF) on the bovine transcriptome in an ex vivo bovine endometrial tissue culture model. Explants were cultured for 30h with sPIF (100nM) or in control media. Total RNA was analysed via RNA-sequencing. As a result of sPIF treatment, 102 genes were differentially expressed compared to the control ( P adj<0.1), although none by more than 2-fold. The majority of genes (78) were downregulated. Pathway analysis revealed targeting of several immune based pathways. Genes for the TNF, NF-κB, IL-17, MAPK and TLR signalling pathways were down-regulated by sPIF. However, some immune genes were demonstrated to be upregulated following sPIF treatment, including C3 . Steroid biosynthesis was the only over-represented pathway with all genes upregulated. We demonstrate that sPIF can modulate the bovine endometrial transcriptome in an immune modulatory manner, like that in the human endometrium, however, the regulation of genes was much weaker than in previous human work.

Preimplantation factor (PIF) is a pregnancy specific peptide with immune modulatory properties exerted on the human endometrium. Viable bovine embryos secrete PIF, but its effect on the bovine endometrial immune response is unknown, both in native and inflammatory stimulated endometrial tissue. An ex vivo bovine endometrial tissue culture model was used with lipopolysaccharide (LPS) as an inflammatory stimulant. The effect of synthetic PIF (sPIF) was assessed, in three separate experiments, on the secretion or mRNA expression of essential prostaglandins and cytokines. Radioimmunoassays were used to assess prostaglandin secretion and ELISA for IL-6 secretion from endometrial explants. mRNA expression of IL6 and IL8 was analysed from endometrial explants with real-time PCR. Synthetic PIF reduced native IL-6 secretion from explants when pre-treated for 24 h. There was no effect of sPIF on IL-6 secretion from LPS challenged explants; however, sPIF increased IL6 mRNA expression when challenged with 500 ng/mL LPS. There was no effect of sPIF on prostaglandin secretion or mRNA expression of IL8. Therefore, sPIF is able to modulate the native IL-6 production pathway in the bovine endometrium, yet demonstrates no effect on prostaglandin secretion or IL8 expression. Unlike in human studies, effects of sPIF were minimal, thus sPIF is not an effective modulator of the immune targets investigated in the bovine endometrium.

ETHNOPHARMACOLOGY RELEVANCE:Maytenus ilicifolia is a Celastracea plant used in traditional medicine to alleviate digestive tract inflammatory disorders. AIM OF THE STUDY:We investigated anti-inflammatory properties of M. ilicifolia crude extract towards Caco-2 cell line, as a model of Toll-like Receptor 2 (TLR-2) inflammatory pathway. MATERIALS AND METHODS:Toxicity was assessed following culture of Caco-2 with M. ilicifolia, using apparent cell permeability and trans-epithelial electric resistance. Anti-inflammatory properties of M. ilicifolia were assessed through IL-8 secretion and TLR-2 associated gene expression of Caco-2 cells with or without an LTA challenge. RESULTS:M. ilicifolia was not toxic to Caco-2 cells. M. ilicifolia down-regulated TLR2 expression with and without LTA challenge but had no effect on other genes. Following LTA challenge of Caco-2 cells, 100 and 200µg/mL M. ilicifolia abrogated IL-8 secretion. CONCLUSIONS:We provide preliminary data for some M. ilicifolia anti-inflammatory properties. Further research must establish the full extent and mode of action on particular inflammatory pathways.

After farrowing piglets are energy deficient. This is enhanced in large litters due to competition for space at the teat and decreased birth weight, for which both factors result in a reduction in piglets’ ability to ingest colostrum. The energy requirements of a new born piglet, needed for movement and maintenance of body temperature, are in excess of the reserves that they are born with. Therefore, the required energy needed in piglets’ early life must come from the limited supply of stored energy they are born with or through feed from the mother. Within the first three days, piglets will receive two types of milk from their mother. Firstly, colostrum, which is produced by the sow before farrowing and secreted up to 12-24 hours after farrowing. Colostrum contains an essential supply of passive immunity and energy that is critical to the piglet in the first 24 hours of its life, both for short and long term survival. Second, transient milk, which is rich in fat and produced by the sow in the intermediary period between colostrum and mature milk, up to 4 days after farrowing. As the newborn piglet is reliant on nutritional and energy input from the sow, the nutrition of the sow during gestation has received considerable interest as a way to improve piglet viability and ultimately survival.

Published in: Advances in Animal Biosciences; Proceedings of the British Society of Animal Science Annual Conference 5 (1): 207