Search Results (523)

Bladder cancer pathogenesis is closely linked to epigenetic alterations, particularly DNA methylation and demethylation processes. Environmental carcinogens and persistent inflammatory stimuli—such as recurrent urinary tract infections—can induce aberrant DNA methylation, altering gene expression profiles and contributing to malignant transformation. This review synthesizes current evidence on the role of DNA methyltransferases (DNMT1, DNMT3a, DNMT3b) and the hypermethylation of key tumour suppressor genes, including A2BP1, NPTX2, SOX11, PENK, NKX6-2, DBC1, MYO3A, and CA10, in bladder cancer. It also evaluates the therapeutic application of DNA-demethylating agents such as 5-azacytidine and highlights the impact of chronic inflammation on epigenetic regulation. Promoter hypermethylation of tumour suppressor genes leads to transcriptional silencing and unchecked cell proliferation. Urine-based DNA methylation assays provide a sensitive and specific method for non-invasive early detection, with single-target approaches offering high diagnostic precision. Animal models are increasingly employed to validate these findings, allowing the study of methylation dynamics and gene–environment interactions in vivo. DNA methylation represents a key epigenetic mechanism in bladder cancer, with significant diagnostic, prognostic, and therapeutic implications. Integration of human and experimental data supports the use of methylation-based biomarkers for early detection and targeted treatment, paving the way for personalized approaches in bladder cancer management. Full article

Leptospirosis is a globally significant zoonosis affecting animal health, productivity, and the environment. While typically associated with tropical climates, its persistence in semi-arid regions such as La Laguna, Mexico—characterized by low humidity, high temperatures, and limited water sources—remains poorly understood. Although these adverse environmental conditions theoretically limit the survival of Leptospira, high livestock density and synanthropic reservoirs (e.g., rodents) may compensate, facilitating transmission. In this cross-sectional study, blood sera from 445 dairy cows (28 herds: 12 intensive [MI], 16 semi-intensive [MSI] systems) were analyzed via microscopic agglutination testing (MAT) against 10 pathogenic serovars. Urine samples were cultured for active Leptospira detection. Risk factors were assessed through epidemiological surveys and multivariable analysis. This study revealed an overall apparent seroprevalence of 27.0% (95% CI: 22.8–31.1), with significantly higher rates in MSI (54.1%) versus MI (12.2%) herds (p < 0.001) and an estimated true seroprevalence of 56.3% (95% CI: 50.2–62.1) in MSI and 13.1% (95% CI: 8.5–18.7) in MI herds (p < 0.001). The Sejroe serogroup was isolated from urine in both systems, confirming active circulation. In MI herds, rodent presence (OR: 3.6; 95% CI: 1.6–7.9) was identified as a risk factor for Leptospira seropositivity, while first-trimester abortions (OR:10.1; 95% CI: 4.2–24.2) were significantly associated with infection. In MSI herds, risk factors associated with Leptospira seropositivity included co-occurrence with hens (OR: 2.8; 95% CI: 1.5–5.3) and natural breeding (OR: 2.0; 95% CI: 1.1–3.9), whereas mastitis/agalactiae (OR: 2.8; 95% CI: 1.5–5.2) represented a clinical outcome associated with seropositivity. Despite semi-arid conditions, Leptospira maintains transmission in La Laguna, particularly in semi-intensive systems. The coexistence of adapted (Sejroe) and incidental serogroups underscores the need for targeted interventions, such as rodent control in MI systems and poultry management in MSI systems, to mitigate both zoonotic and economic impacts. Full article

Leptospirosis is a (re-)emerging and global zoonotic disease. Given the complex host-pathogen interaction and the numerous environmental risk factors related to the transmission, a One Health approach to both disease prevention and control is needed. Occurring at the human–cattle–environment interfaces, bovine leptospirosis represents a zoonotic risk for the professionals in the field, besides being a potential cause of significant economic losses due to the bovine reproductive disorders. Although climatic change is a potential factor in exacerbating the risk of leptospirosis in Europe, this disease remains largely neglected, with several knowledge gaps in research, investigations, and diagnosis of bovine genital leptospirosis syndrome across the continent. The present report describes the results of the diagnostic investigations on a case of chronic bovine leptospirosis in a breeding bull. Following the seroconversion to Leptospira Sejroe var Hardjo after the arrival of the animal in a quarantine facility, a monitoring plan including both serological/molecular analyses and a therapeutic protocol was undertaken. The bull exhibited a persistent seroconversion and a repeated positivity for Leptospira to real-time PCR in urine samples, indicative of a chronic shedder pattern. This report emphasizes the diagnostic and management challenges in the context of such a complex but frequently overlooked disease. Full article

A key factor in calculating dairy cows’ nitrogen (N) excretion is knowing the amount of daily excreted urine. The present study aimed to investigate two methods to calculate the daily urine volume (UV) excreted using spot urine samples. Data were obtained from nine balance experiments involving 47 lactating and seven non-lactating German Holstein cows, with an average body weight (BW) of 620 ± 95 kg and an average age of 5.6 ± 1.4 years. Daily urinary creatinine (Cr) and UVs were known for all animals. The first method was developed by linearly regressing the daily excreted amount of Cr in urine against BW (p < 0.001; R² = 0.51; RSE: 2.8). The slope of the regression was used to calculate UV. The second method includes a non-linear regression of UV on Cr concentration in urine, allowing direct estimation of UV without knowledge of BW (p < 0.001; RSE: 8.13). Both estimation methods were compared to the standard method to determine UV from balance trials using Lin’s concordance correlation coefficient (CCC) and Bland–Altman plots. The first method had a CCC of 0.81, and the second method had a CCC of 0.85. Both methods can confidently be applied to calculate UV. Therefore, the second method is usable if BW is unavailable. Full article

Urine-derived and plant-derived benzoic acid can accumulate within soil, and it likely changes the soil microbial community as well as N₂O emissions; however, its mechanism is not clear. This study conducted an incubation experiment to monitor N₂O emissions under low moisture (40% water-filled pore space (WFPS)) and high moisture (85% WFPS) conditions. Metagenomic sequencing and q-PCR methods were used to determine the link between N₂O emissions and the composition and functions of soil microbiota. Benzoic acid (BA) was found to significantly, yet dose-dependently, impact N₂O emissions; that is, low BA concentrations increased N₂O, whereas high BA decreased N₂O. However, this was only found under high moisture conditions. In contrast, BA had little impact on N₂O emissions under low moisture conditions. Under high moisture conditions, BA increased the gene copy number of bacteria and fungi, and decreased the ratio of bacteria to fungi. Similarly, BA significantly increased the abundance of denitrification functional genes, but reduced the (NirS + NirK)-to-NosZ ratio at the peak of emission. This is in agreement with the observation of the increased relative abundance of genes encoding N₂O reductase (EC 1.7.2.4) and NO₃⁻ heterotrophic reductase (EC 1.7.1.15, EC 1.7.2.2) in the metagenomic analysis. In summary, high concentrations of benzoic acid reduce N₂O emissions by promoting the reduction of N₂O. This study revealed the impact of BA on soil microbiota and highlighted the favorable conditions and underlying mechanism behind BA’s significant impact on soil N₂O emissions. This study’s novelty lies in the fact that it deepens our understanding of the complicated role of root exudates and metabolites of animals and plants in soil. Full article

The appearance rate of nutrients into systemic circulation affects hormones like insulin and through that efficiency of growth. This also affects mineral requirements critical for metabolism, notably phosphate (P), magnesium (Mg), and potassium (K). Fasting animals have a downregulated metabolism, upon which P, Mg, and K are exported from their cells into the blood and are subsequently excreted in their urine. Abrupt resumption of feed intake, especially of highly glycemic feeds, creates an acute need for these minerals, which can result in deficiency symptoms, particularly with P deficiency. In human medicine, this is called refeeding syndrome: a large meal after a period of fasting can prove fatal. Young animals seem to be especially sensitive, likely driven by their ability to grow rapidly and thus to drastically upregulate their metabolism in response to insulin. Symptoms of P deficiency are fairly a-specific and, consequently, not often recognized. They include edema, which makes it appear as if piglets are growing well, explaining the high gain/feed rate typically seen immediately after weaning, even when piglets are eating at or below the maintenance requirements. Phosphate deficiency can also result in hypoxia and hypercarbia, which may trigger ear necrosis, Streptococcus suis infections, or even death. Hypophosphatemia can also trigger rhabdomyolysis, which may contribute to tail-biting, but this requires further study. Arguably, when fasting cannot be avoided, diets for newly weaned piglets should be formulated to avoid these problems by lowering their glycemic load and by formulating diets according to the piglets’ actual requirements inspired by their genuine intake and health and not simply by extrapolating from older animals. Full article

Resorcylic acid lactones (RALs) are fungal metabolites with known biological activity. Zeranol, a synthetic RAL, has been used as an estrogenic growth promoter in cattle; however, its use is prohibited in several countries. Zearalenone, a mycotoxin produced by Fusarium spp., is commonly found in contaminated animal feed and can be metabolized into other RALs, which are subsequently excreted in urine. To differentiate between natural contamination from feed and the illegal administration of zeranol, the European Union Reference Laboratory for Growth Promoters (EURL) developed a mathematical equation. This study aims to evaluate the detection and quantification of RALs in bovine urine from animals fed zearalenone-contaminated diets, implanted with zeranol, or subjected to both conditions. RALs were detected and quantified in the urine of cattle consuming contaminated corn, while zeranol and taleranol were identified in the urine of implanted animals. The EURL equation proved to be a valuable tool for determining the origin of RALs in bovine urine and holds significant potential for monitoring and enforcing regulations regarding the illegal use of zeranol. Full article

Background/Objectives: Herbal extracts from Betula alba (birch) are traditionally used for their purported diuretic effects, but scientific evidence supporting these claims remains limited. In this pilot study, we evaluated the short-term effects of a standardized B. alba leaf extract in healthy adult rats using an untargeted urinary metabolomics approach based on UPLC-QTOF. Methods: Two doses, 25 or 50 mg/kg, of a standardized B. alba extract were orally administered to rats. The extract contains hyperoside (0.53%), quercetin glucuronide (0.36%), myricetin glucoside (0.32%), and chlorogenic acid (0.28%) as its main constituents. After 3 days of treatment, the 24 h urine output was measured. Results: While no statistically significant changes were observed in the 24 h urine volume or the urinary Na⁺ and K⁺ excretion, multivariate metabolomic analysis revealed treatment-induced alterations in the urinary metabolic profile. Notably, the levels of two glucocorticoids, i.e., corticosterone and 11-dehydrocorticosterone, were increased in treated animals, suggesting that the extract may influence corticosteroid metabolism or excretion, potentially impacting antidiuretic hormone signaling. Elevated bile-related compounds, including bile acids and bilin, and glucuronidated metabolites were also observed, indicating changes in bile acid metabolism, hepatic detoxification, and possibly gut microbiota activity. Conclusions: Although this study did not confirm a diuretic effect of B. alba extract, the observed metabolic shifts suggest broader systemic bioactivities that warrant further investigation. Overall, the results indicate that the approach based on urinary metabolomics may be valuable in uncovering the mechanisms of action and evaluating the bioactivity of herbal extracts with purported diuretic properties. Full article

Encephalitozoon cuniculi is a microsporidian widely spread in rabbits (Oryctolagus cuniculus) and other species, including humans, causative of neurological disorders or remaining in a latent state in the host organism. The aim of this study was to estimate the prevalence of E. cuniculi in rabbits located in the North-Western region of Romania, and to run a comparative diagnosis for E. cuniculi by multiple methods. A total of 381 rabbits were included, originating from households, family farms and wildlife, which were subjected to serological and/or molecular genetics diagnostic methods for E. cuniculi identification. Seropositivity of 43.02% (151/351) was obtained by ELISA, together with a 45.45% (110/242) prevalence from urine, feces and organs by nested PCR. Additionally, a prevalence of 48.39% (15/31) was identified by a comparative real-time PCR (qPCR). The urinary bladder is firstly reported for molecular E. cuniculi diagnosis, with a positivity of 30.56% (11/36) by nested PCR. Despite the disagreement between the diagnostic methods, the present results highlight the level of pathogen dissemination among rabbits in North-Western Romania that represents a risk for not only rabbits and other animals, but also for the general public by its zoonotic character. Full article

Mycotoxins are synthesized by various fungal species and are known to exert toxic effects on vertebrates and other animals, even at low concentrations. However, the current countermeasure for mycotoxin contamination is random inspection of samples prior to shipment. In this study, we focused on Chlorella sorokiniana (CS) from Ishigaki Island, Japan, and examined its ability to detoxify deoxynivalenol (DON), ochratoxin A (OTA), and fumonisin B1 (FB1) in vitro and in vivo. The binding of CS to DON, OTA, and FB1 was evaluated in vitro. The detoxification of CS was demonstrated by monitoring its concentrations in the plasma and urine samples of male ICR mice. Plasma and urine samples were collected 30 min, 2 h, and 24 h after an oral administration of 5 mg/kg mycotoxins and/or 500 mg/kg CS. CS bound to more than 80% and 40% of DON and OTA, respectively, whereas the binding of CS to FB1 was less than 10%. The concentrations of DON and OTA in plasma and urine samples were substantially reduced by CS co-administration, whereas CS did not affect FB1 absorption. The co-administration of CS substantially inhibited the systemic absorption of DON and OTA. Full article

Objective: This study aimed to examine the impact of high-fat-diet(HFD)-induced obesity on pain sensitivity and tetrahydrobiopterin (BH4) levels. The effect of moderate-intensity physical exercise, an anti-inflammatory non-pharmacological intervention, on pain scores was also investigated. Methods: Adult male C57BL/J6 mice were fed standard or an HFD for eight weeks. Their total body weight, food intake, locomotor and motivational behavior, and pain reflexes were measured. A subgroup of animals underwent physical exercise for five days per week over six weeks. Blood was collected for glucose tolerance testing and levels of lactate. Urine samples were collected to measure BH4 levels. Results: We showed that the HFD increased weight gain, epididymal white adipose tissue, and the percentage of body weight as epididymal fat. These anthropometric alterations were characterized by impaired glucose tolerance at four and eight weeks of the dietary intervention. It was also observed that reduced locomotor activity and higher pain scores in the HFD-fed mice were prevented by a physical exercise intervention. The HFD also induced an increase in urinary BH4 levels at four and eight weeks of intervention. Conclusions: The HFD increased scores of chemical and mechanical hyperalgesias, as well as urinary BH4 levels. Urinary BH4 can be proposed as a potential easy-to-access, sensitive, and reliable biomarker of pain development, and a promising target for the control of pain hypersensitivity in obesity. Full article

Tactical athletes and military personnel engaged in intense exercise need to consume enough quality protein in their diet to maintain protein balance and promote recovery. Plant-based protein sources contain fewer essential amino acids (EAAs), while pork loin contains a higher concentration of EAAs and creatine than most other animal protein sources. This study aimed to determine whether the ingestion of plant-based or pork-based military-style meals ready-to-eat (MREs) affects recovery from and subsequent Army Combat Fitness Test (ACFT) performance. Methods: Twenty-three (n = 23) University Corps of Cadets members participated in a randomized, double-blind, placebo-controlled, and crossover-designed study. Diets were prepared by a dietitian, food scientist, and chef to have similar taste, appearance, texture, and macronutrient content. The chef also labeled the meals for double-blind administration. Participants refrained from intense exercise for 48 h before reporting to the lab in a fasted condition with a 24 h urine sample. Participants donated a blood sample, completed questionnaires and cognitive function tests, and consumed a pre-exercise meal. After four hours, participants performed the ACFT according to military standards. Participants were fed three MREs daily while returning to the lab in a fasted condition at 0600 with 24 h urine samples after 24, 48, and 72 h of recovery. On day 3, participants repeated the ACFT four hours after consuming an MRE for breakfast. Participants resumed normal training and returned to the lab after 2–3 weeks to repeat the experiment while consuming the alternate diet. Data were analyzed using general linear model statistics with repeated measures and percent changes from baseline with 95% confidence intervals. Results: Results revealed that 3 days were sufficient for participants to replicate ACFT performance. However, those consuming the pork-based diet experienced less muscle soreness, urinary urea excretion, cortisol, inflammation, and depression scores while experiencing a higher testosterone/cortisol ratio and appetite satisfaction. There was also evidence of more favorable changes in red and white blood cells. Conversely, blood lipid profiles were more favorably changed when following a plant-based diet. Conclusions: These findings suggest that protein quality and the availability of creatine in the diet can affect recovery from intense military-style exercise. Minimally, plant-based MREs should include 6–10 g/d of EAA and 2–3 g/d of creatine monohydrate to offset dietary deficiencies, particularly in military personnel following a vegetarian diet. Registered clinical trial #ISRCTN47322504. Full article

The increasing number of owned and stray dogs in large cities is becoming a pressing issue due to rising population densities, urban conditions, and poor control over animal reproduction. This situation poses serious epidemiological risks, as dogs can act as reservoirs and transmitters of infectious and parasitic diseases dangerous to humans. This study aimed to investigate the prevalence and carriage of infectious and parasitic diseases in stray dogs in the city of Uralsk as a factor of epidemiological risk. In 2024, 1213 stray dogs were captured from different city districts and examined at the veterinary clinic and laboratory of Zhangir Khan University. Biological samples (blood, urine, feces) from 10% of the animals were analyzed using molecular (PCR), serological (ELISA), and helminthological methods. Serological and molecular analyses revealed the widespread circulation of bacterial pathogens. Antibodies to additional bacterial agents, including Pasteurella multocida, Mycobacterium spp., Listeria monocytogenes, and Leptospira spp., were detected in the samples, indicating an unfavorable sanitary and epidemiological situation in the urban environment. An enzyme-linked immunosorbent assay (ELISA) identified antibodies against Toxocara canis in 50.9% of the dogs and against Echinococcus granulosus in 76.4%, reflecting both active and past infections. The polymerase chain reaction (PCR) results showed the presence of Brucella canis DNA in blood and urine samples, while antibodies to Brucella spp. were detected in 57.8% of the examined dogs, underscoring the significant zooanthroponotic importance of this pathogen and its potential threat to human health. Additionally, T. canis DNA was found in 39.2% of the samples and E. granulosus DNA in 16.6%. A helminthological examination using the Fülleborn method revealed a high rate of helminth infection: Ancylostoma caninum—35.3%, T. canis—32.3%, and Toxascaris leonina—29.4%. The obtained results highlight the significant role of stray dogs as epizootiological and epidemiological reservoirs of zooanthroponotic infections. This poses a serious threat to public health and necessitates the implementation of effective control and prevention measures for infectious and parasitic diseases within urban fauna. Full article

The aim was to investigate the suitability of Santhica 27 industrial hemp leaves as a protein source in dairy cow nutrition. Twelve Holstein dairy cows received a total mixed ration (TMR) containing 7.4% industrial hemp leaves (HEMP) and a TMR containing 3.5% soya extraction meal (CON) in a crossover design. Cows were kept in a free-stall barn for 2 weeks to measure feed intake, milk yield and sample plasma, ruminal fluid, and urine. In week 3, cows were housed in a respiration chamber to measure gas exchange, urine, and feces excretions. In the first two weeks, cows of the HEMP group rested longer but spent less time ruminating. Feeding the HEMP diet reduced dry matter intake (DMI), milk yield, and urinary N-metabolite concentrations and tended to lower total N-excretion, milk fat, and lactose concentrations. During the stay in the respiration chamber, DMI, milk yield, apparent digestibility, and crude protein degradability were similar between groups, but feeding the HEMP diet tended to reduce methane yield. In conclusion, Santhica 27 hemp leaves are a suitable protein source for dairy cows as they have no negative effects on animal health, apparent digestibility, and crude protein degradability. Nevertheless, inadequate adaptation to the diet reduces feed intake and milk yield. Full article

In recent years, much attention has been paid to environmental protection, not only by reducing emissions of harmful gases from industry, but also by reducing the excretion of biogenic compounds or ammonia emissions from agriculture, including animal production. The aim of this study was to determine the effects of complete diets with reduced inclusion levels of crude protein and limiting essential amino acids, fed to pigs in two- and three-phase feeding systems, and the feeding system on crude protein digestibility, nitrogen retention and utilization, fecal and urinary pH, fecal and urinary nitrogen and ammonia levels, and nitrogen excretion. Digestibility-balance trials were performed on 24 growing–finishing pigs housed in individual metabolism crates, in three groups, in two- and three-phase feeding systems. The pigs were fed the following diets: C-control diet; L-low-protein diet where the levels of crude protein and essential amino acids (lysine, methionine + cystine, threonine, and tryptophan) were reduced by 15% relative to diet C; L+AA-low-protein diet supplemented with crystalline lysine, methionine, threonine, and tryptophan to the standard levels (as in diet C). Diets L fed to pigs in two- and three-phase feeding systems significantly decreased crude protein digestibility and nitrogen retention, particularly in the three-phase system. The supplementation of diets L+AA with crystalline essential amino acids improved crude protein digestibility and nitrogen retention and utilization, especially in the two-phase system. Reduced concentrations of crude protein and essential amino acids in diets L contributed to a significant increase in feces and urine acidity in both two- and three-phase feeding systems. The supplementation of diets L+AA with essential amino acids resulted in a significant increase in urinary pH and a non-significant increase in fecal pH. This experimental factor had no effect on fecal ammonia concentration in group L+AA. The values of pH and total fecal nitrogen were somewhat higher in the two-phase system than in the three-phase system. Fecal ammonia concentration was similar in both systems. The three-phase feeding system contributed to a decrease in urinary pH and total urinary nitrogen. The analyzed feeding systems had no significant effect on urinary ammonia concentration. It was estimated that a reduction in crude protein (by 20–25 g/kg) and essential amino acid levels in pig diets, relative to the standard levels, reduced nitrogen excretion by 18.7% and 15.6% in two- and three-phase feeding systems, respectively. The supplementation of low-protein diets (L) with lysine, methionine, threonine, and tryptophan induced a further reduction in nitrogen excretion. A comparison of the effects of feeding systems (two-phase system vs. three-phase system) on crude protein digestibility and nitrogen retention and utilization revealed that better results were obtained in the two-phase feeding system. Full article