IntroductionMaternal nutrition during pregnancy critically influences offspring development and immune function. One-carbon metabolites (OCM) are epigenetic modifiers that may modulate antimicrobial peptide (AMP) expression, which is vital for innate immunity. This study investigated the effects of maternal nutrient restriction and OCM supplementation on mRNA expression of AMP in fetal and maternal lung, mammary gland, and small intestine of beef cattle.MethodsTwenty-nine crossbred Angus beef heifers were synchronized for estrus and artificially inseminated. They were assigned to one of four treatments in a 2 × 2 factorial design: nutritional plane [control (CON) vs. restricted (RES)] and OCM supplementation [without OCM (−OCM) or with OCM (+OCM)]. Heifers on the CON diet were fed to gain 0.45 kg/day, while RES heifers were fed to lose 0.23 kg/day. Treatments were applied from day 0 to 63 of gestation, after which all heifers were fed a common diet to gain 0.45 kg/day until day 161 of gestation, when samples were collected. Quantitative RT-qPCR was used to assess mRNA expression of AMP.ResultsNutritional plane had no effect (p ≥ 0.24) on mRNA expression of AMP in either the fetus or dams. However, the mRNA expression of cathelicidin5 (CATHL5; p = 0.07) and bovine neutrophil β-defensin5 (BNBD5; p = 0.07) in the fetal lung and mammary gland, respectively, was lower in the +OCM groups compared to the −OCM groups. In the maternal small intestine, the expression of enteric β-defensin (EBD) was lower (p = 0.01) in the +OCM groups compared to the −OCM groups. Additionally, in the maternal lung, there was a tendency (p = 0.06) for an interaction in CATHL5 mRNA expression, with the RES + OCM group showing greater expression compared to the CON + OCM (p = 0.07) and RES − OCM (p = 0.08) groups.DiscussionOur findings suggest that while restricted maternal nutrition did not affect mRNA expression of AMP, OCM supplementation modulated AMP expression in both fetal and maternal tissues. Further research is needed to elucidate the mechanisms underlying OCM’s impact on AMP expression.

Abstract Background During transportation many horses develop post‐transportation infection, which can be life‐threatening and end their sport career. Preventing mucus accumulation and inflammation during transportation is vital, emphasizing the need for effective strategies to enhance overall horse health welfare. Objectives Assess the impact of N‐acetylcysteine (NAC) on mucus accumulation and inflammation in horses subjected to 18 hours of head confinement. Animals Six healthy crossbred horses, 5.3 ± 2.1 years of age and weighing 387 ± 30 kg. Methods Prospective placebo‐controlled cross‐over design study. The horses' heads were restrained in their stalls for a period of 18 hours. They were studied under 4 conditions: Not confined (NC): before head confinement, placebo (P), and confined head (CH): 18 hours of head confinement without treatment, and N‐Acetylcysteine (NAC): 18 hours of head confinement treated with NAC before confinement (15 mg/kg/day NAC PO for 3 days). Bronchoalveolar lavage (BAL) was performed in each condition. Mucus accumulation along the trachea was evaluated by endoscopy. Results Endoscopic scores were significantly different between CH and other conditions, whereas no significant differences were found among NC, P, and NAC. The BAL cell count (34 291 ± 2624 cells/μL), neutrophil and lymphocyte count (18 601 ± 3193 cells/μL and 3337.4 ± 593 cells/μL, respectively) in CH were significantly higher compared to NAC. Neutrophil percentage was significantly higher in CH (53.8 ± 8%) compared to horses that received NAC (20.08 ± 8%). Conversely, in comparison to NAC (66.33 ± 9%), the percentage of macrophages was significantly lower in CH (35.7 ± 10%). Conclusions N‐acetylcysteine was found to significantly decrease mucus accumulation and inflammatory cell counts in horses with head confinement.

To investigate the effects of nutrient restriction and one-carbon metabolite (OCM) supplementation (folate, vitamin B12, methionine, and choline) on fetal small intestine weight, vascularity, and cell proliferation, 29 (n = 7 ± 1 per treatment) crossbred Angus beef heifers (436 ± 42 kg) were estrous synchronized and conceived by artificial insemination with female sexed semen from a single sire. Then, they were allotted randomly to one of four treatments in a 2 × 2 factorial arrangement with the main factors of nutritional plane [control (CON) vs. restricted feed intake (RES)] and OCM supplementation [without OCM (−OCM) or with OCM (+OCM)]. Heifers receiving the CON level of intake were fed to target an average daily gain of 0.45 kg/day, which would allow them to reach 80% of mature BW by calving. Heifers receiving the RES level of intake were fed to lose 0.23 kg/heifer daily, which mimics observed production responses in heifers that experience a diet and environment change during early gestation. Targeted heifer gain and OCM treatments were administered from d 0 to 63 of gestation, and then all heifers were fed a common diet targeting 0.45 kg/d gain until d 161 of gestation, when heifers were slaughtered, and fetal jejunum was collected. Gain had no effect (p = 0.17) on the fetal small intestinal weight. However, OCM treatments (p = 0.02) displayed less weight compared to the −OCM groups. Capillary area density was increased in fetal jejunal villi of RES − OCM (p = 0.02). Vascular endothelial growth factor receptor 2 (VEGFR2) positivity ratio tended to be greater (p = 0.08) in villi and was less in the crypts (p = 0.02) of the RES + OCM group. Cell proliferation decreased (p = 0.02) in villi and crypts of fetal jejunal tissue from heifers fed the RES + OCM treatment compared with all groups and CON − OCM, respectively. Spatial cell density increased in RES − OCM compared with CON + OCM (p = 0.05). Combined, these data show OCM supplementation can increase expression of VEGFR2 in jejunal villi, which will promote maintenance of the microvascular beds, while at the same time decreasing small intestine weight and crypt cell proliferation.

Background:Injectable products, particularly human vaccines, must be free from fever-inducing agents and thoroughly tested for pyrogens as part of a quality control. Consequently, manufacturing facilities are required to conduct ppropriate pyrogen tests per pharmacopoeial standards. This study aimed to evaluate the reliability of the monocyte activation test (MAT) in quantifying pyrogenic content in the recombinant hepatitis B vaccine. Methods:We assessed pyrogen activity in the active pharmaceutical ingredient (API), formulated vaccine, and aluminum hydroxide by comparing the limulus amebocyte lysate (LAL), rabbit pyrogen test (RPT), and MAT, measuring activity in relative pyrogen unit (RPU) as per the European Pharmacopoeia. Monocytes from healthy donors were isolated and identified via flow cytometry to measure the CD14+ marker frequency. Results:The study found that the pyrogenic concentration of lipoteichoic acid (LTA) in the MAT was 50,000 ng/mL (5.19 EEU/mL). In contrast, the same concentration in the rabbit pyrogen test (RPT) was deemed non-pyrogenic based on rectal temperature assessments. The MAT showed sensitivity to the API and adjuvant, with a detection limit of 2.5 EU/mL for IL-6, outperforming the RPT, which had a detection limit of 5 EU/mL. Conclusion:A strong IL-6 response to both lipopolysaccharide (LPS) and LTA stimulation was observed, indicating that IL-6 could serve as a valuable marker for pyrogen testing. The MAT appears to be an effective alternative to the RPT for assessing pyrogenicity, demonstrating commendable consistency and accuracy across various testing systems allowed by the Ph. Eur. General MAT Chapter, especially given the RPT's limitations in controlling pyrogenicity in injectable products.

The current study aimed to evaluate the effects of parturition and different stages of milk production on Cardiac Troponin I activity (CTnI) and other cardiac markers. A total of 42 healthy dairy cattle with an average body weight of 520 +/- 17 kg and an average parity of 2.4 +/- 0.7 were randomly selected from a commercial dairy farm. Through a clinical examination and ECG recording conducted by a veterinarian, blood samples were collected from all cattle's coccygeal vein one month before parturition (dry period), one week after parturition (early lactation), and 10 weeks after parturition (peak of the lactation). Dairy cows in the experimental group were observed closely before and during parturition and in the postpartum period. Biochemical parameters, including Cardiac Troponin I activity (CTnI), Creatine kinase (CK), Aspartate aminotransferase (AST), and Lactate dehydrogenase (LDH) were analyzed at three-time intervals. Regarding time intervals, there were no significant differences among the cattle in terms of CTnI, CK, LDH, AST, parturition, and different stages of milk production. It was found that the effect of late-stage gestation and different lactation periods on cardiac biomarkers in dairy cows was not significant.

Objective of our study were to investigate the impact of maternal nutrition during early pregnancy on the vascularity and proliferation of fetal intestine and whether supplementing one-carbon metabolites (OCM) could mitigate any changes arising from altered maternal nutrition. We used a 2 x 2 factorial arrangement of treatments with two levels of heifer gain: control (CON; 0.45 kg/d) and restricted (RES; -0.23 kg/d), with and without (+/- OCM) supplementation. Crossbred Angus beef heifers were estrous synchronized and bred via artificial insemination with single-sire female-sexed semen. Heifers were divided into four nutritional treatments (n = 8 ± 1 per treatment) at breeding. Targeted heifer gain (achieved by changing in intake of a common diet) and OCM treatments were administered from d 0 to 63 of gestation and then all heifers were fed a common diet targeting 0.45 kg/d gain. Treatments containing OCM were given rumen-protected methionine (10 g/d) and choline (60 g/d) in a fine-ground corn carrier administered daily, with weekly injections of folate (320 mg) and vitamin B12 (20 mg). The non-supplemented treatment (-OCM) received the corn carrier and saline injections. On day 161 of gestation heifers were harvested and samples were collected from fetal jejunum for analysis. Vascularity was assessed via measurement of capillary density using anti-CD31 and anti-CD34 fluorescent staining. Cell proliferation within intestinal tissues was assessed by KI-67 fluorescent staining. Images were analyzed using ImagePro-Premiere software. Maternal gain × OCM interactions were not present (P > 0.11). Fetuses from restricted dams had greater (P = 0.05) capillary area density in the villi compared with CON (5.48 vs. 4.72 ± 0.38%), while OCM had no effect. Crypt and total cell proliferation were greater (P < 0.05) in CON. Providing OCM reduced (P = 0.01) total intestinal cell proliferation. The observed increase in capillary density in the villi of fetuses from restricted dams may be a compensatory response to reduced maternal nutrient availability, aimed at preparing the offspring for increasing post-natal nutrient delivery. Conversely, the decreased proliferation in crypt cells appears to be a logical response to restricted nutrition and may indicate a slowing of the rate of cell division, which could have long-term consequences for intestinal development. Our results also demonstrate that maternal OCM can reduce total intestinal cell proliferation in the fetal intestine, regardless of maternal nutrition. One-carbon metabolites have crucial roles in DNA methylation and other cellular processes for proper cellular function and development. Observed decreases resulting from the provision of OCM may indicate enhanced energetic efficiency, reduced epithelial cell turnover, and/or reduce apoptotic events in the villa. Further studies are needed to determine the long-term effects of maternal OCM on fetal intestinal development and function. USDA is an equal opportunity provider and employer.

Abstract Six 100‐day‐old mixed‐breed lambs were examined in a farm with a semi‐intensive system due to neurologic signs. Cachexia, bilateral blindness, stupor, severe drowsiness and lethargy with left and right movements of the head and neck were recorded after awakening and stimulation. Lambs died 10 days after the onset of the clinical signs. The lambs were necropsied, and after routine parasitology, bacteriology and histopathology, the occurrence of acute coenurosis was confirmed due to finding multiple cystic structures in the brain tissue. All lambs of the herd were treated with albendazole (orally, 25 mg/kg, two doses with an interval of 14 days). All shepherd dogs were treated with popantel (orally, one tablet/10 kg, two doses with an interval of 14 days). The affected lambs died despite this treatment. No new case of the disease was observed after the initiation of control measures. The present study shows the importance of preventive measure against coenurosis in a semi‐intensive sheep farming system that includes implementing consistent parasite control programme in dogs being in contact with sheep.

β-Defensins are cationic antimicrobial peptides (AMPs) that play an important role in the innate immune defense of bovines. They are constitutively expressed in mammary glands and induced differently in response to pathogens. Their expression is influenced by various factors, including hormones, plant-derived compounds, and dietary energy imbalance. The toll-like receptors (TLRs)/nuclear factor-kappa B (NF-κB) pathway plays a crucial role in β-defensin induction, while alternative pathways such as mitogen-activated protein kinase (MAPK) and epigenetic regulation also make substantial contributions. β-Defensins exhibit bactericidal activity against a wide range of pathogens, including two major mastitis pathogens, Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), primarily through membrane disruption. β-Defensins have low cytotoxicity to host cells and demonstrate immunomodulatory properties, and pathogens also display minimal resistance to these AMPs. Given the increasing concern in antimicrobial resistance, the potential of β-defensins as natural antimicrobials has garnered considerable attention. This article provides an overview of the characteristics of bovine β-defensins, their expression pathways, their mode of action, and factors influencing their expression in the mammary glands of cattle. Additionally, it identifies the current gaps in research within this field and suggests areas that require further investigation. Understanding the regulation and function of β-defensins offers valuable insights to develop effective strategies for strengthening the immune system of mammary glands, reducing the reliance on synthetic antimicrobials, and explore novel natural antimicrobial alternatives.