Animals, Volume 14, Issue 6 (March-2 2024) – 166 articles

The objective of this study was to develop an in vitro model that mimics inflammatory reactions and neutrophil extracellular traps (NETs) formation by polymorphonuclear leukocytes (PMNs) in dairy cows. This model was used to examine the effect of carprofen (CA) on lipopolysaccharide (LPS)-induced NETs formation and expression of inflammatory factors. Peripheral blood samples were collected from 24 Holstein cows (3–11 days postpartum) and PMNs were isolated. In three replicates, PMNs were exposed to various treatments to establish an appropriate in vitro model, including 80 μg/mL of LPS for 2 h, followed by co-incubation for 1 h with 60 μmol/L CA and 80 μg/mL LPS. The effects of these treatments were evaluated by assessing NETs formation by extracellular DNA release, gene expression of pro-inflammatory cytokines, reactive oxygen species (ROS) production, and the expression of NETs-related proteins, including histone3 (H3), citrullinated histone (Cit-H3), cathepsin G (CG), and peptidyl arginine deiminase 4 (PAD4). The assessment of these parameters would elucidate the specific mechanism by which CA inhibits the formation of NETs through the PAD4 pathway instead of modulating the Nox2 pathway. This highlights CA’s effect on chromatin decondensation during NETs formation. Statistical analyses were performed utilizing one-way ANOVA with Bonferroni correction. The results demonstrated that LPS led to an elevated formation of NETs, while CA mitigated most of these effects, concurrent the PAD4 protein level increased with LPS stimulating and decreased after CA administration. Nevertheless, the intracellular levels of ROS did not change under the presence of LPS. LPS supplementation resulted in an upregulation of H3 and Cit-H3 protein expression levels. Conversely, the CA administration inhibited their expression. Additionally, there was no change in the expression of CG with either LPS or LPS + CA co-stimulation. The gene expression of pro-inflammatory cytokines (tumor necrosis factor -α, interleukin (IL)-18, IL-1β, and IL-6) upregulated with LPS stimulation, while the treatment with CA inhibited this phenomenon. In conclusion, CA demonstrated a pronounced inhibitory effect on both LPS-induced NETs formation as well as the associated inflammatory response. Full article

This longitudinal case study provides an in-detail report of the process towards the elimination of Mycobacterium avium subsp. paratuberculosis (MAP) from a closed 450-head commercial dairy herd. In parallel, two diagnostic approaches were applied to all cows in annual intervals during 2012–2022: detection of MAP in individual faecal samples by bacteriological cultivation on solid medium and detection of MAP-specific antibodies by ELISA. For each annual sampling, the kappa coefficients for test agreement and the survival rates of MAP-positive and MAP-negative cows were calculated. Applying a multivariable linear regression model revealed a significantly lower fat-corrected 305-day milk yield for MAP-positive cows. The true prevalence of MAP shedders reduced from 24.2% in 2012 to 0.4% in 2019 and during 2020–2022, no MAP shedder was identified. Test agreement was generally low and bacteriological cultivation showed positive results earlier than the ELISA. In the first years of control, the survival of MAP shedders was longer than in the final stage. In conclusion, the elimination of MAP from a dairy herd might be feasible within a decade. Changes in the test agreement must be considered. Timely removal of MAP shedders, hygienic calf rearing, and colostrum supply are key for successful control. Full article

Viral Encephalopathy and Retinopathy (VER) is a neurological infectious fish disease that causes vacuolization and necrosis in the central nervous system, which lead to swimming abnormalities and, generally, host death in the early stages of development. VER is caused by the Nervous Necrosis Virus (NNV), a non-enveloped virus with a bisegmented and positive-stranded (+) RNA genome. The largest segment (RNA1) codes for viral polymerase while capsid protein is encoded by RNA2. The aim of this study was to explore the potential of a reverse-engineered RGNNV/SJNNV strain that harbors mutations in both 3′NCRs (position 3073 of RNA1 and 1408 and 1412 of RNA2) as an attenuated live vaccine for sole. The attenuation of this strain was confirmed through experimental infections in sole at 22 °C. Vaccination trials were performed by bath, intramuscular, and intraperitoneal injection, at two temperatures (18 and 22 °C). Our results indicate the improved survival of vaccinated fish and delayed and poorer viral replication, as well as an overexpression of immune response genes linked to T cell markers (cd4 and cd8), to an early inflammatory response (tlr7 and tnfα), and to antiviral activity (rtp3 and mx). In conclusion, our study indicates that the attenuated strain is a good vaccine candidate as it favors sole survival upon infection with the wt strain while inducing a significant immune response. Full article

The western-European hedgehog (Erinaceus europaeus), in expanding its range towards human habitats, faces exposure to contaminants and biological agents, potentially leading to diseases associated with hematological and biochemical changes. As bioindicators of environmental pollution and carriers of zoonotic agents, hedgehogs play a crucial role in One Health studies, emphasizing the need for a comprehensive understanding of their clinical-pathological aspects. Exploring the blood reference values in healthy animals of this species is crucial for understanding and improving their well-being, and identifying possible diseases/pathogens that may affect its conservation and/or impact human health. This review is focused on analyzing the data available in the literature for Erinaceus europaeus blood reference intervals. A comprehensive literature review of the studies published in Europe is performed, highlighting their specificities, and emphasizing the need for continuous research in this field. Our final goal is to provide a crucial tool for assessing the health status of the species, and underscoring the significance of research in this specific domain. Full article

The current study investigates the venom-delivery system of green and red morphotypes of the sea anemone Actinia equina to disclose its potential as a source of bioactive compounds. We compared the two morphotypes using electron and optical microscopy, proteomics, and toxicity assessment on zebrafish embryos. Specialized venom-injecting cells (nematocysts) are equally distributed and found in the tentacles of both varieties. Proteomics revealed proteins of interest in both red and green Actinia, yielding the three most abundant Gene Ontology (GO) terms related to the biological processes “proteolysis”, “hemolysis in another organism” and “lipid catabolic process”. Neurotoxins and cytolytic toxins similar to known cnidarian toxins like PsTX-60A and AvTX-60A, for instance, were identified in both types. Extracts from green and red anemones were toxic to zebrafish embryos, with green anemone venom appearing to be more potent. The findings highlight the presence of proteinaceous toxins in A. equina and the potential for different varieties to possess distinct bioactive compounds. Notably, pore-forming toxins are suggested for molecular probes and immunotoxins, making them valuable assets for potential biotechnological and biomedical purposes. Full article

Neoschoengastia gallinarum is widely distributed in Asia, preferentially parasitising birds, and heavy infestations have clinical impacts on domestic fowl. In common with other trombiculid mites, the genetic diversity and potential variation in host preferences or pathology induced by N. gallinarum are poorly understood. This study aimed to unravel the geographical variation and population structure of N. gallinarum collected from galliform birds in Peninsular Malaysia and Thailand by inference from concatenated mitochondrial-encoded cytochrome c oxidase subunit I (COI), and nuclear-encoded internal transcribed spacer 2 (ITS2) and 18S ribosomal DNA gene sequences, including a comparison with previously published data from southeastern China. Our multi-locus sequence analysis revealed three monophyletic clades comprising (A) specimens from Peninsular Malaysia, (B) the samples from Thailand together with a minority of Chinese sequences, and (C) the majority of sequences from China. Similarly, most species delimitation approaches divided the specimens into three operational taxonomic units. Analysis of molecular variance revealed 96.41% genetic divergence between Malaysian and Thai populations, further supported by the absence of gene flow (Nm = 0.01). In conclusion, despite the two countries sharing a land border, populations of N. gallinarum from Peninsular Malaysia and Thailand appear to be genetically segregated and may represent distinct cryptic species. Full article

A prior investigation revealed that a lack of Zinc (Zn) could hinder intestinal cell proliferation in broiler chickens; however, the mechanisms responsible for this effect remain unclear. We aimed to investigate the possible mechanisms of dietary Zn deficiency in inhibiting the jejunal cell proliferation of broilers. For this study, a total of 112 chickens (21 days old) were randomly divided into two treatments (seven replicate cages per treatment, eight chickens per replicate cage): the control group (CON) and the Zn deficiency group. The duration of feeding was 21 d. Chickens in the control group were provided with a basal diet containing an extra addition of 40 mg Zn/kg in the form of Zn sulfate, whereas chickens in the Zn deficiency group were given the basal diet with no Zn supplementation. The results indicated that, in comparison to the CON, Zn deficiency increased (p < 0.05) the duodenal and jejunal crypt depth (CD) of broilers on d 28 and jejunal and ileal CD on d 35, and decreased (p < 0.05) the duodenal, jejunal, and ileal villus height/crypt depth (VH/CD) on d 28 and the jejunal VH, jejunal and ileal villus surface area, and VH/CD on d 35. Furthermore, Zn deficiency decreased (p < 0.0001) the number of proliferating cell nuclear antigen-positive cells and downregulated (p < 0.01) the mRNA or protein expression levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K, phosphorylated serine–threonine kinase (AKT), phosphorylated mechanistic target of rapamycin (mTOR), G protein-coupled receptor 39 (GPR39), and extracellular-regulated protein kinase, but upregulated (p < 0.05) the mRNA or protein expression levels of P38 mitogen-activated protein kinase, c-jun N-terminal kinase (JNK) 1 and JNK2, and phosphorylated protein kinase C in the jejunum of the broilers on d 42. It was concluded that dietary Zn deficiency inhibited cell proliferation possibly via the GPR39-mediated suppression of the PI3K/AKT/mTOR signaling pathway in the jejunum of broilers. Full article

Flight is a complex physiological process requiring precise coordination of muscular contraction. A key protein in insect flight is flightin, which plays an integral role in the flight muscles. This research sought to evaluate the flight competence of the social wasp V. basalis by characterizing the molecular components involved. Our study focused on Vespa basalis, one of the most dangerous hornet species, utilizing PCR to obtain a partial cDNA sequence of the flightin protein. We then employed phylogenetic and sequence analysis to gain insights into this protein in flight-related adaptations. The cDNA has an 1189-base pair sequence including an open reading frame (453 bp) encoding 150 amino acids. Analyzing the deduced amino acid sequence using an online tool revealed a molecular weight of 18.05 kDa, an isoelectric point of 5.84, four functional site patterns, and no transmembrane topology. We constructed a phylogenetic tree of flightin based on 38 species. Our analysis indicated that V. basalis is most closely related to V. mandarinia; this alignment is consistent with their similar aggressive behavior, but their evolutionary relationship, based on mitochondrial sequences, presents a contrast. These initial findings on the flightin gene in V. basalis lay the groundwork for future functional studies to elucidate its specific role in flight adaptations and explore its potential as a target for pest management strategies. Full article

This study aimed to assess the effective tissue concentrations of the current standard of care for pain mitigation in calves during castration (injectable lidocaine) and to assess the ability of a lidocaine-loaded elastration band (LLB) to deliver effective concentrations into the scrotal tissue over time. This study comprised two different trials: (1) effective concentrations of injectable lidocaine in the scrotal tissue; and (2) the in vivo delivery of effective concentrations of lidocaine from LLBs placed on the calf scrotums. Sensation in the scrotal tissue was assessed by electrocutaneous stimulation. Injectable lidocaine allowed for short-term anesthesia for up to 60 min, highlighting the importance of finding additional strategies to mitigate long-term pain. An elastomeric ligation band impregnated with lidocaine could provide a suitable alternative, as it yielded tissue levels of lidocaine that approached EC₅₀ and exceeded EC₉₅ at 2 and 72 h following application, respectively, and remained above those levels for at least 28 days after application. Further studies are warranted to compare the use of LLBs to injectable local anesthetics. Full article

The Boerboel dog breed (BBD) is indigenous to South Africa (SA) and plays an important role in safeguarding homes and farms. The Department of Agriculture, Land Reform, and Rural Development (DALRRD) classifies the BBD as a protected species, and it is valued for its intelligence, boldness, and strength, as well as for continually ensuring the safety of its owners. The aim of this study was to investigate genetic diversity within the BBD population using pedigree information. The original BBD data, which contained 87,808 records, were obtained from the Integrated Registration and Genetic Information System (INTERGIS). After editing, the pedigree data included 87,755 records of animals born between 1971 and 2019. Pedigree analyses were performed using PEDIG (Fortran 77 software) to determine the completeness, inbreeding coefficients, and genetic diversity as defined by the genetic contributions of the most important ancestors of the current animals. This study identified 91.2% inbred animals in the BBD population, with an average and maximum inbreeding of 7.5% and 50% of inbred animals, respectively. The estimated inbreeding rate per year was 0.20% with an effective population size of 83.1. The most influential ancestors explained 82.63% and 80.92% of the total genetic variation for males and females in the studied populations, respectively. Only 10 important ancestors explained more than 50% of the entire population’s genetic diversity. The numbers of founders ($f$) were 348 and 356, and the effective numbers of founders ($f_e$) were 57.4 and 60.1, respectively, for males and females. The numbers of founders were higher than the effective numbers of founders, implying a loss of genetic diversity due to unequal founder contributions. The BBD population was not critically endangered based on the inbreeding rates and effective population size; however, the population experienced a significant loss of genetic variability, unequal genetic contributions by founders, and a genetic bottleneck. Future breeding strategies could benefit from using equal proportions of parent stock and including new genetically distant breeds. Full article

In twin pregnancies of discordant sex, the male fetus grows larger than the female co-twin. Our study aimed to determine the effect of the sex of co-twins on lambs’ birth weight in ovine pregnancies developed under natural undernourishment. Additionally, we investigated whether the nutritional and/or antioxidant supplementation provided to ewes during pregnancy could modulate the potential effects associated with the sex of co-twins. Ninety-six birth records of twin pregnancies of sheep grazing the natural Patagonian prairies were analyzed. The animals were divided into four groups: control (no supplementation), N (concentrate supplementation, 100% NRC), A (antioxidant supplementation), and NA (concentrate + antioxidant supplementation). Supplementation occurred from day 35 of gestation onwards until lambing. There were no differences in female or male birth weight in the control undernourished group. However, in group N, females or males with sex-discordant co-twins had a higher birth weight than did those with co-twins of the same sex. Group A males with female co-twins had a higher birth weight compared to males whose co-twins were also males. In NA lambs, males had a higher birth weight compared to females, regardless of their co-twin’s sex. Therefore, chronic undernutrition abolished the differences in birth weight due to fetal sex. Restoring maternal nutrition or antioxidant supplementation tends to normalize birth weight and restore the differences between females and males. This effect is enhanced with the combined supplementation of concentrated food and antioxidants. Full article

Exposure to heat stress (HS) detrimentally affects pig performance. This study explored whether a dietary phytogenic solution based on Capsicum spp. (PHY) could enhance the thermal tolerance of heat-stressed growing pigs. Forty-two individually housed pigs were randomly assigned to three treatments: thermoneutral pigs on a control diet (TN-C) and pigs subjected to HS fed the control diet either without (HS-C) or with supplemental PHY (HS-PHY). The TN-C group exhibited increased average daily gain (ADG) and feed intake (FI) compared to both HS-C (p < 0.01) and HS-PHY pigs (p < 0.05) and better feed efficiency compared to HS-C pigs only (p < 0.01). However, the HS-PHY pigs showed significantly higher FI (p < 0.01) and ADG (p < 0.05) compared to HS-C pigs. HS pigs displayed higher body temperatures (BTs) than TN pigs (p < 0.01), yet HS-PHY pigs experienced a lesser increase in BT compared to HS-C pigs (p < 0.05). Supplementation with PHY mitigated some effects of HS, increasing serum superoxide dismutase and catalase activity, reducing HSP90 expression in longissimus dorsi muscle, and elevating jejunal villus height compared to HS-C pigs (p < 0.05), reaching levels akin to TN-C pigs. Additionally, PHY supplementation resulted in lower serum urea levels than HS-C pigs (p < 0.01) and similar myosin gene expression to TN-C pigs (p > 0.1), suggesting enhanced amino acid post-absorptive utilization for lean tissue growth. In conclusion, dietary PHY supplementation partially offset the adverse effects of HS on pig performance by improving thermal tolerance. Full article

Animal welfare is inherited in each society, shaped by the surrounding environment and upbringing of each individual. This influence becomes particularly significant during adolescence. Due to its cultural context, Spain is among the European Union nations with the highest number of cases of animal abuse. The Canary Islands, the scenario of this study, show the highest rates of intentional poisoning of wildlife and pets’ abandonment. The aim of the present study was to explore the perception of animal welfare among adolescents, studying the influence of the main socio-demographic factors that may condition it. A validated questionnaire on animal abuse was used and distributed to adolescents aged 14–18 years in two public study centers. Animal abuse rates were correlated with socio-demographic variables. In total, 302 subjects answered the questionnaire. The perception of animal welfare was influenced by socio-demographic variables, gender being the most important. The demographic profile of the least responsive adolescent to animal abuse was a male engaged in sports, not owning a dog, and hailing from a family involved in hunting. Awareness should be raised at an early age, promoting artistic activities, encouraging contact with animals and sporting practices that do not generate a lack of empathy for animals. Full article

This study investigated the effects of a synbiotic consisting of inulin, Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis, and Lactobacillus reuteri given orally to day (d)-of-hatch (DOH) broiler chicks at the hatchery and in the feed for a 21 d period. A total of 480 Cobb male broilers were randomly divided into one of four treatments using a 2 × 2 factorial design as follows: (1) control (CTRL) group receiving a gel-only oral application on DOH at the hatchery prior to transport and a non-medicated basal corn/soybean meal starter diet; (2) hatchery synbiotic (HS) receiving an oral gel containing the synbiotic (0.5 mL/bird) at the hatchery and the basal diet; (3) CTRL + dietary synbiotic at 0.5 kg/MT (DS); and (4) HS + dietary synbiotic at 0.5 kg/MT (HSDS). On d 7 and d 21, one bird per pen (eight replicate pens/group) was euthanized, and the ileum was immediately removed for qPCR analysis. Data were subjected to a 2-way ANOVA using GLM procedure (JMP Pro17). A significant diet × hatchery interaction was observed in feed conversion ratio (FCR) from d 14 to d 21 (p = 0.013) where the HS, DS, and HSDS treatments had a significantly lower FCR compared to the CTRL. However, no significant interaction effect was observed for body weight gain (BWG) or FCR during the overall experimental period. No significant interaction was observed in mRNA abundance of the evaluated genes in the ileum on d 7 and d 21. Gel application with the synbiotic significantly reduced sodium-dependent glucose cotransporter 1 (SGLT1) mRNA abundance on d 7 (p = 0.035) in comparison to birds receiving gel alone. Regardless of hatchery application, dietary synbiotic supplementation significantly reduced Toll-like receptor (TLR)2, TLR4, and interleukin (IL)-10 mRNA abundance on d 7 (p = 0.013). In conclusion, these findings showed that hatchery and dietary synbiotic application could have a potential beneficial impact on broiler intestinal immunity by regulating the TLR response, a key element of innate immunity. FCR was improved from d 14 to d 21 after synbiotic application. Future research involving extended grow-out studies with a disease challenge would expand on the implications of an early application of synbiotics. Full article

Parapoxviruses are worldwide epitheliotropic viruses that affect ruminants. Viruses of this genus have a narrow host range; however, the pseudocowpox virus (PCPV) also infects humans. Unfortunately, these cases are not well documented, and the epidemiology and the properties of the causative agents are not properly described. Here, we report the first case of PCPV in northern Russia (the Irkutsk region). The infection occurred in non-immune herds where no new arrivals of animals had been reported. Moreover, clinical signs of infection (skin lesions) were observed in humans. Based on the nucleotide identity and phylogenetic analysis of the partial-length B2L gene, the Irkutsk 2019 isolate was classified as PCPV. Phylogenetic analysis based on the nucleotide sequence of the B2L gene fragment of PCPV revealed a close phylogenetic relationship between the Irkutsk 2019 isolate and the PCPV strains isolated in Europe and the USA. The high degree of conservatism of the B2L gene does not allow for finding a correlation between their geographical origin and the results of phylogenetic analysis. Full article

Birds that roam freely, particularly those that migrate, have the potential to carry a range of diseases that can be passed on to humans. The vast movement of these birds across diverse environments and urban areas can contribute to the spread of bacteria over long distances, impacting both human and animal populations. Stress, overcrowding, and human interaction can also play a role in the transmission of infectious diseases among birds and humans. Therefore, it is crucial to comprehend the intricate connections between birds, vectors, zoonotic pathogens, and the environment, especially given the increasing urbanization and emergence of zoonotic illnesses. This review aims to provide a systematic overview of the significance of avian species in transmitting bacterial pathogens that pose a risk to public health. Full article

The utilisation of seaweeds as feed supplements has been investigated for their potential to mitigate enteric methane emissions from ruminants. Enteric methane emissions are the primary source of direct greenhouse gas emissions in livestock and significantly contribute to anthropogenic methane emissions worldwide. The aim of the present study is to evaluate the nutritional role and the in vitro effect on cumulative gas and methane production of Asparagopsis taxiformis (native species) and Asparagopsis armata (invasive species), two species of red algae from the Azorean Sea, as well as the ability to reduce biogas production when incubated with single pasture (Lolium perenne and Trifollium repens) as substrate. Four levels of concentrations marine algae were used (1.25%, 2.25%, 5%, and 10% DM) and added to the substrate to evaluate ruminal fermentation using the in vitro gas production technique. The total amount of gas and methane produced by the treatment incubation was recorded during 72 h of incubation. The results indicate that both algae species under investigation contain relatively high levels of protein (22.69% and 24.23%, respectively, for Asparagopsis taxiformis and Asparagopsis armata) and significant amounts of minerals, namely magnesium (1.15% DM), sodium (8.6% DM), and iron (2851 ppm). Concerning in vitro ruminal fermentation, it was observed that A. taxiformis can reduce enteric methane production by approximately 86%, during the first 24 h when 5% is added. In the same period and at the same concentration, A. armata reduced methane production by 34%. Thus, it can be concluded that Asparagopsis species from the Azorean Sea have high potential as a protein and mineral supplement, in addition to enabling a reduction in methane production from rumen fermentation. Full article

Small-scale carp polyculture plays a key role in food supply in Bangladesh. However, factors including water pollution, limited infrastructure, and inadequate disease management hinder its sustainability. This paper reports on a survey of 231 farmers across the six major carp producing regions in Bangladesh, analyzing factors including farmers’ social aspects, farm characteristics, information on disease and approaches adopted to combat them, and biosecurity practices. Almost half (46.8%) of the farms surveyed experienced disease in carp species, with clear regional variations. Eighty-four percent of farms reported carp mortalities during disease outbreaks, with an average mortality level of 10.23 ± 11.81%. Clinical signs during outbreaks lasted between a week and a month, with a peak in disease outbreaks occurring in two seasonal periods between June and July and October and December. Disease incidence was related to a range of factors including the farmer’s experience, ponds/farm type, stocked species, and biosecurity practice. A combination of disinfecting measures during pond preparation and measures during stocking, including discarding fingerling transport water away from the farm, fingerling disinfection, and checking the health of fingerlings before stocking, significantly reduced disease occurrence. Treatments involving antibiotics, ciprofloxacin, erythromycin, and azithromycin were reported as ineffective, raising concerns about their non-prudent use, inadequate dosing (perhaps without appropriate veterinary guidance), and the potential for driving antimicrobial resistance in the environment. The research unveils a concerning pattern of high disease incidence across small-scale carp farms in Bangladesh, and the significant potential for disease spread highlights the need for responsible disposal practices. The study emphasizes the need for improving training and awareness programs for addressing biosecurity and disease management challenges, ensuring sustainable aquaculture and community well-being. Full article

Uninfected alligators (Alligator mississippiensis) exhibited high constitutive levels of hepatic gene expression related to immune function, whereas the highest-expressed hepatic genes of uninfected mice were related to metabolism. Intraperitoneal challenge of mice with bacterial lipopolysaccharide results in dramatic inflammatory effects including peritoneal ascites, febrile response, dramatic alterations in electrophoretic serum profile, and mortality. In contrast, coelomic injection of alligators with 200× the murine LD₅₀ of intraperitoneal bacterial lipopolysaccharide resulted in no changes in serum protein profiles, behavioral effects, mortality, and no coelomic ascites. However, injection of juvenile alligators with live bacteria resulted in a titer-dependent decrease in metabolic rate, as measured by oxygen consumption. These results are the opposite of those observed for mammalian and avian species. The decreased oxygen consumption was not accompanied by changes in heart or respiration rate, indicating that this phenomenon was not due to bradycardia or bradypnea. Interestingly, challenge of alligators with bacteria resulted in the complete expulsion of digestive tract contents within four hours. We interpret these activities as temporary minimization of other biological systemic activities to redirect and devote energy to immune function. The reallocation of resources within an organism to fight infection without increases in metabolic rate has not been described in other animals. Full article

This paper describes a model for the prediction of methane and ammonia emissions from fattening pig houses. This model was validated with continuous and discrete measurements using a reference method from two manure management systems (MMS): long storage (LS) in deep pits and short storage (SS) by daily flushing of a shallow pit with sloped walls and partial manure dilution. The average calculated methane and ammonia emissions corresponded well with the measured values. Based on the calculated and measured results, the average calculated CH₄ emission (18.5 and 4.3 kg yr⁻¹ per pig place) was in between the means from the continuous data from sensors (15.9 and 5.6 kg yr⁻¹ per pig place) and the means from the discrete measurements using the reference method (22.0 and 3.1 kg yr⁻¹ per pig place) for the LS and SS systems, respectively. The average calculated NH₃ emission (2.6 and 1.4 kg yr⁻¹ per pig place) corresponded well with the continuous data (2.6 and 1.2 kg yr⁻¹ per pig place) and the discrete measurements using the reference method (2.7 and 1.0 kg yr⁻¹ per pig place) from LS and SS, respectively. This model was able to predict the reduction potential for methane and ammonia emissions by the application of mitigation options. Furthermore, this model can be utilized as a predictive tool, enabling timely actions to be taken based on the emission prediction. The upgraded model with robust calculation rules, extensive validations, and a simplified interface can be a useful tool to assess the current situation and the impact of mitigation measures at the farm level. Full article

Histopathologic examination of intestinal biopsies from dogs with acute hemorrhagic diarrhea syndrome (AHDS) reveals necrotizing enteritis and epithelial integrity loss. Serum iohexol measurement has been utilized to assess intestinal permeability. Our hypothesis is that dogs with AHDS have increased intestinal permeability, which is associated with the severity of clinical signs. In this prospective case–control study, 53 client-owned dogs (28 AHDS, 25 healthy controls) were evaluated. Clinical severity was assessed using the AHDS index and systemic inflammatory response syndrome (SIRS) criteria. Simultaneously, dogs received oral iohexol, and serum iohexol concentrations (SICs) were measured two hours later. Results indicated significantly higher (p = 0.002) SIC in AHDS dogs (median: 51 µg/mL; min–max: 9–246) than in healthy controls (30 µg/mL; 11–57). There was a significant positive correlation between AHDS index and SIC (r_S = 0.4; p = 0.03) and a significant negative between SIC and serum albumin concentrations (Pearson r = −0.55; p = 0.01). Dogs with severe AHDS (mean 106 µg/mL; range: 17–246) demonstrated significantly higher (p = 0.002) SIC than those with mild to moderate disease (29 µg/mL; 9–54). These findings underscore the association between intestinal permeability and clinical severity in dogs with AHDS assessed by iohexol. Full article

The Chinese soft-shelled turtle (Pelodiscus sinensis) is an important freshwater aquaculture turtle due to its taste and nutritional and medicinal value. More ecological culturing modes, such as rice–turtle co-culture, should be developed to meet the ecological benefit demand. We compared growth, physiological parameters, and transcriptome data to detect the physiological responses and regulatory mechanisms of pond-cultured turtles as compared to co-cultured turtles. The co-cultured turtles grew slower than pond-cultured turtles. The gonadosomatic index of co-cultured male turtles was lower than that of pond-cultured male turtles, and both the mesenteric fat index and limb fat index were lower in co-cultured turtles than in pond-cultured turtles (p < 0.05). The blood GLU of the co-cultured turtles was significantly lower than the GLU of the pond-cultured turtles (p < 0.05), while the values of CRE, UA, BUN, AKP, ACP, GOT, and CAT were higher in the co-cultured turtles than in the pond-cultured turtles (p < 0.05). In total, 246 and 598 differentially expressed genes (DEGs) were identified in the brain and gut from turtles cultured in the two different modes, respectively. More DEGs were related to environmental information processing, metabolism, and human diseases. In the brain, the top enriched pathways of DEGs included the longevity regulating pathway, glycerolipid metabolism, cytokine–cytokine receptor interaction, Toll-like receptor signaling pathway, and PI3K-Akt signaling pathway, while in the gut, the top enriched pathways of DEGs included the cell cycle, DNA replication, cellular senescence, and p53 signaling pathway. The turtles acclimated to the different culturing conditions by adjusting their growth, physiological, and biochemical characteristics and related gene expression during a short culture period. Full article

In recent years, nitrate (NO₃-N) pollution in water bodies has been increasing due to the excessive use of nitrogen-based fertilizers. Exposure to NO₃-N during the development of amphibian embryos may have lasting effects on the growth and development of individuals and even threaten their survival, but the toxicity mechanism of NO₃-N in amphibian embryos prior to thyroid morphogenesis remains unclear. In the present study, Bufo gargarizans was selected as the model organism to investigate the toxic effects of 10 mg/L and 100 mg/L NO₃-N exposure (N10 and N100) on amphibian embryos using methimazole (MMI) and exogenous thyroxine (T4) as the reference groups. We found that T4, MMI, N10 and N100 inhibited B. gargarizans embryo growth and development, with MMI and N100 showing the earliest and strongest effects. Transcriptome analysis revealed that MMI and NO₃-N (especially N100) significantly downregulated genes related to thyroid morphogenesis and cholesterol metabolism, while upregulating genes related to inflammation and apoptosis. Together, these results contribute to a deeper understanding of the complex mechanisms by which NO₃-N disrupts B. gargarizans embryonic development, reveal the potential risks of NO₃-N pollution to other aquatic organisms, and provide insights into the conservation of a broader ecosystem. Full article

Feed supplements supporting animal welfare and performance are becoming increasingly important. Immunomodulatory effects of such products have been observed in many species. The aim of this study was to analyze whether food supplementation with a Saccharomyces cerevisiae fermentation product (SCFP) affects the occurrence of foal diarrhea in early life, and whether the SCFP feeding has an impact on the immediate response to a parenteral vaccination at the age of 6–9 months. Eleven foals received the SCFP (OLI) and eleven foals were fed a placebo (PLA) for 29 days. Growth, diarrhea, and diarrhea severity were observed until day 30. After weaning, at the age of 6–9 months, foals were vaccinated parenterally against influenza and tetanus. The supplementation had no statistically significant effect on diarrhea duration and severity. On the day of vaccination, PLA and OLI foals did not differ significantly regarding numbers of circulating blood leukocyte subsets. However, the response to vaccination differed significantly between OLI and PLA foals. In OLI foals, the numbers of the major leukocyte fractions (granulocytes, lymphocytes, monocytes, CD4+ T cells, CD8+ T cells, CD21+ B cells, and MHC-II+/CD21− cells) increased significantly 24 h after vaccination but remained unchanged in PLA foals. The observed results suggest that early life supplementation with an SCFP may affect the early immune response to an initial vaccination. Full article

To investigate the effects of individual essential amino acids (EAA) on growth and the underlying mechanisms, EAA individually supplemented a low-protein (LP) diet fed to young rats in the present study. Treatments were an LP diet that contained 6% crude protein (CP), a high-protein (HP) diet that contained 18% CP, and 10 LP diets supplemented with individual EAA to achieve an EAA supply equal to that of the HP diet. The CP concentration of the LP diet was ascertained from the results of the first experiment, which examined the effects of dietary CP concentrations on growth rates, with CP ranging from 2% to 26%. Weight gain was increased with the supplementation of His, Ile, Lys, Thr, or Trp as compared to the LP diet (p < 0.05). Feed intake was greater for the His-, Lys-, and Thr-supplemented treatments as compared to the LP group (p < 0.05). Protein utilization efficiency was lower for the HP group than other groups (p < 0.01). The supplementation of Leu, Lys, and Val led to reduced protein utilization efficiency (p < 0.05), but the supplementation of Thr and Trp led to greater efficiency than the LP group (p < 0.05). Compared to the LP group, plasma urea concentrations were elevated with individual EAA supplementation, with the exception of the Thr addition. The added EAA resulted in increased concentrations of the corresponding EAA in plasma, except for Arg and Phe supplementation. The supplementation of Arg, His, Leu, Lys, and Met individually stimulated mTORC1 pathway activity (p < 0.05), and all EAA resulted in the decreased expression of ATF4 (p < 0.05). In summary, the supplementation of His, Ile, Lys, Thr, or Trp to an LP diet improved the growth performance of young rats. Responses to His and Lys additions were related to the activated mTORC1 pathway and feed intake increases. The improved growth performance resulting from the addition of a single EAA is not solely attributed to the increased plasma availability of EAA. Rather, it may be the consequence of a confluence of factors encompassing signaling pathways, the availability of amino acids, and other associated elements. The additivity of these factors results in independent responses to several EAA with no order of limitation, as is universally encoded in growth models for all production animal species. Full article

Sixteen 35-day-old male crossbred rabbits (New Zealand white × Thai native breed) with an initial weight of 484 ± 11.3 g were randomly divided into two groups of eight, constituting control and treatment groups. The treatment group was orally administered a crude extract of butterfly pea (Clitoria ternatea L.) at 0.5 g/kg body weight from weaning (at 35 days) to slaughter (at 90 days). The effects on the phagocytic activity of blood polymorphonuclear leukocytes, serum biochemistry, meat quality, muscular lipid peroxidation, the apparent digestibility of dry matter and nutrients, and gut histology were studied. The results revealed that the phagocytic function of circulating leukocytes (75 and 90 days) and alveolar macrophages (90 days) did not differ between the two groups. At slaughter, treated rabbits had lower blood urea nitrogen concentrations and higher liver weight than control rabbits (p < 0.05). After chilling at 4 °C for 24 h, a lower meat pH and the alteration of meat color (brighter, less yellow, lower hue angle, and decreased color saturation) were observed in the treated group (p < 0.05). Furthermore, lipid peroxidation (measured at 3, 5, and 7 storage days) in the meat of treated rabbits was lower than in controls (p < 0.05). The apparent digestibility of organic matter and ether extract (analyzed at 46 days for 4 days) was improved in the treated group (p < 0.05), whereas gut histology was unaffected. In conclusion, butterfly pea extract supplementation did not affect phagocytic function but led to a modification in meat color, delayed lipid peroxidation, and improved digestibility. Full article

Aeromonas schubertii is a pathogen that severely affects aquatic animals, including the snakehead, Channa maculata. Lytic bacteriophages have been recognized as effective alternatives to antibiotics for controlling bacterial infections. However, there have been no reports of A. schubertii phages as far as we know. In this study, a lytic bacteriophage SD04, which could effectively infect A. schubertii, was isolated from pond water cultured with diseased snakehead. The SD04 phage formed small, round plaques on Petri dishes. Electron microscopy revealed a hexagonal head and a contractile tail. Based on its morphology, it may belong to the Myoviridae family. Two major protein bands with molecular weights of 50 and 38 kilodaltons were observed after the phage was subjected to SDS-PAGE. The phage showed a large average burst size, high specificity, and a broad host range. When stored at 4 °C, phage SD04 had high stability over 12 months and showed almost no variation within the first six months. All fish were healthy after both intraperitoneal injection and immersion administration of SD04, indicating the safety of the phage. After treatment with SD04, Channa maculata in both phage therapy groups and prevention groups showed high survival rates (i.e., 83.3 ± 3.3% and 100 ± 1.3%, respectively). Phage therapy inhibits bacterial growth in the liver, the target organ of the infected Channa maculat. The experimental results indicate the potential use of phage SD04 for preventing A. schubertii infection in Channa maculata. Full article

This study aimed to evaluate whether supplemental feeding at milking (SFAM) positively influences the quantitative−qualitative milk parameters due to improving some welfare assessment traits of multiparous Anatolian buffalo cows confined in semi-open free-stall barns. A total of 76 Anatolian buffalo cows at approximately 90 days in milk were selected to encompass four groups (OSF-2nd, NSF-2nd, OSF-≥3rd and NSF-≥3rd), considering offering (OSF) or not (NSF) supplemental feed at milking and the parity (2nd) and (≥3rd). Data of evaluated variables such as the following ones—(i) subjectively scored welfare assessment traits (temperament, udder hygiene and body condition), (ii) milk yield per milking (MYM), (iii) milk components, and (iv) milk physical traits—were analysed using a linear mixed model and principal component (PC) analysis. The OSF improved the temperament, udder hygiene and body condition scores compared to the NSF. The MYM, the fat content and the fat-to-protein ratio of the OSF were higher than those of the NSF, but milk mineral and electrical conductivity of the OSF were lower than those of the NSF. The parity of cows did not affect the evaluated variables. Four parameters (milk density value and lactose, solids-not-fat and protein contents) could be identified in the PC2 versus PC1 plot. In conclusion, the SFAM enhanced the milk yield and qualitative milk parameters due to improving the welfare status of indoor buffalo cows, regardless of parity. Full article