Animal Physiology Research Articles

Systemic Changes in Early Pregnancy in the Mare: An Integrated Proteomic Analysis of Blood Plasma, Histotroph, and Yolk Sac Fluid at Day 14 Post‐Ovulation

ABSTRACTPurposeEmbryo‐maternal signaling during the establishment of pregnancy in horses remains one of the biggest mysteries in large animal physiology. Early pregnancy loss represents a major source of economic loss to the breeding industry. This study aimed to investigate the systemic changes associated with early pregnancy by mapping the proteome of blood plasma at 14 days in pregnant and non‐pregnant mares.Experimental DesignPlasma proteomes were analysed in commercially bred pregnant (n = 17) and non‐pregnant (n = 17) Thoroughbred mares at 14 days after ovulation, using high‐resolution mass spectrometry. Day 14 histotroph and yolk sac fluid were also profiled and datasets were integrated through pathway analysis.ResultsWe identified 229 total protein IDs, with 12 increased and 10 decreased significantly in pregnant versus non‐pregnant plasma. To gain functional insight, these data were aligned with proteomes of 14‐day pregnant mare uterine fluid (n = 4; 1358 IDs) and conceptus fluid (soluble proteins within the yolk sac fluid; n = 4; 1152 IDs), and further interrogated using gene ontology databases and pathway analysis.Conclusions and Clinical RelevanceThese analyses identified consistent systemic changes in the mare's proteome that indicate a profound and specific immune response to early pregnancy, which appears to precede the systemic endocrine response to pregnancy. Integrated pathway analysis suggests that embryo‐maternal interactions in early pregnancy may mimic elements of the virus‐host interaction to modulate the maternal immune response. Transthyretin (TTR) and uteroglobin (SCGB1A1) were respectively down‐ and upregulated in plasma while also present in uterine fluid, and are proposed to be key proteins in early pregnancy establishment. These findings contribute significantly to our knowledge of early pregnancy in the mare and identify potential new avenues for developing clinical approaches to reduce early embryo loss.

Metabolic Scaling: Exploring the Relation Between Metabolic Rate and Body Size.

We present an alternative to the traditional classroom lecture on the topics of metabolic scaling, allometric relationships between metabolic rate (MR) and body size, and reasons for rejecting Rubner''s surface "law," concepts that students have described as challenging, counterintuitive, and/or mathematical. In groups, students work with published data on MR and body size for species representing all five vertebrate groups. To support the exercise, we developed a worksheet that has students define the concept in their own words, compare different measures of MR, and evaluate plots of MR and mass-specific MR versus body mass for both homeotherms and poikilotherms. Students also attempt to explain why selected species have exceptionally high or low MR values for their body sizes. Student feedback indicated active learning is an effective way to learn the concepts of metabolic scaling and allometric relationships and that the opportunity to work in groups with real data stimulates interest and an appreciation for the importance of metabolic scaling to the understanding of animal physiology.

Understanding the Lymphatic System: Tissue-on-Chip Modeling.

The lymphatic vasculature plays critical roles in maintaining fluid homeostasis, transporting lipid, and facilitating immune surveillance. A growing body of work has identified lymphatic dysfunction as contributing to the severity of myriad diseases and to systemic inflammation, as well as modulating drug responses. Here, we review efforts to reconstruct lymphatic vessels in vitro toward establishing humanized, functional models to advance understanding of lymphatic biology and pathophysiology. We first review lymphatic endothelial cell biology and the biophysical lymphatic microenvironment, with a focus on features that are unique to the lymphatics and that have been used as design parameters for lymphatic-on-chip devices. We then discuss the state of the art for recapitulating lymphatic function in vitro, and we acknowledge limitations and challenges to current approaches. Finally, we discuss opportunities and the need for further development of microphysiological lymphatic systems to bridge the gap in model systems between lymphatic cell culture and animal physiology.

Reviving diversity: cryoprotectants and culturing methods enhance recovery of mammalian gut microbes from field samples

Abstract The field of microbial ecology is increasingly recognizing the need for methods to isolate and culture gut microbes to better understand how these microorganisms impact animal physiology, especially in mammalian hosts. Currently, there is a lack of clear methods to store microbial samples for cultivability, especially when samples are collected from the field, transported to the laboratory, and preserved under long-term storage for weeks to months compared to mere days in the biomedical field. Here, the cecal contents of groundhogs (Marmota monax) were processed and stored with or without various preservation solutions at −80 °C for at least 2 months. All microbial samples were then grown in distinct nutrient media in liquid and plate conditions and were incubated under anaerobic and aerobic environments. Treatment comparisons revealed that the samples stored in preservation solutions containing 1 or more cryoprotectants provided the greatest and most consistent bacterial densities. To test the long-term storage efficacy of the preservation solutions, we inventoried taxonomic identities and abundances of these cultures using 16S rRNA amplicon sequencing. Our findings highlight that: (1) preserved samples containing cryoprotectants exhibited the highest microbial richness and diversity and resembled the original cecal samples the most when grown under anaerobic conditions; and (2) the effect of individual animal identity was detectable in the membership of cultured communities, irrespective of preservation solutions. Our study is the first to demonstrate the importance of preservation solutions containing multiple cryoprotectants for long-term storage and further microbial culturing and novel isolation. Understanding and improving storage methods that preserve microbial physiology and conserve their compositional diversity is essential for field-collected samples useful in mammalian microbiome and culturomics studies, promoting a better comprehension of the identity and function of wild host-associated microbiomes.

The Effect of Drinking Ionized Water on the Productive Performance, Physiological Status, and Carcass Characteristics of Broiler Chicks.

Water treatment technologies have received great attention recently, as water is the most important nutritional element, and animals consume it daily in larger quantities than those of food. The ideal water treatment affects the chemical composition and physical properties of water, having a significant positive impact on the animal's physiology, productivity, and welfare. Studies conducted on water ionization devices for broiler chickens remain limited; therefore, this study was planned to investigate the effect of ionized drinking water on the productive performance, physiological status, and carcass characteristics of broiler chicks. A total of 900 one-day-old broiler chicks were randomly and equally assigned to three groups, each with six replicates (50 birds/replicate). The first group (C) received tap drinking water and served as a control, while the second group (T1) received ionized drinking water from an ionizing device that worked for 1 h/100 L. The third group (T2) received ionized drinking water from an ionizing device that worked for 2 h/100 L. Water analysis for each treatment was performed. Productive traits, such as weekly body weight, feed intake, and water intake, were recorded. Hematological parameters and biochemical constituents were measured according to the reference's description. Furthermore, carcass characteristics, such as carcass weight and dressing percentage, and bacterial count of the intestine, such as Lactobacilli and Coliform counts, were determined. From the results, ionized water (T1 and T2) had a negative ORP, which is often desirable as it suggests the presence of antioxidant properties and lower total dissolved solids (TDSs), heterotrophic plate count (HPC), and algal total count (ATC) than in tap water. The treated chicks showed higher final body weights and better feed conversion rates than the control. Ionized water also improved carcass quality characteristics, such as carcass weight and dressing percentage. T1 and T2 chicks exhibited higher hemoglobin, total protein, globulin, G and M immunoglobulin, and total antioxidant capacity (TAC) levels, as well as lower malondialdehyde (MDA) and low-density lipoprotein (LDL) levels than the control. Furthermore, they had lower pathogenic bacteria counts. Therefore, it is recommended to employ the ionizing approach for broiler chicken drinking water, particularly a 2 h/100 L ionization application, for better animal productivity, health, and welfare.

What a tangled web we weave: crosstalk between JAK-STAT and other signalling pathways during development in Drosophila.

The Janus kinase-signal transducer and activator of transcription (JAK-STAT) signalling pathway is a key player in animal development and physiology. Although it functions in a variety of processes, the net output of JAK-STAT signalling depends on its spatiotemporal activation, as well as extensive crosstalk with other signalling pathways. Drosophila, with its relatively simple signal transduction pathways and plethora of genetic analysis tools, is an ideal system for dissecting JAK-STAT signalling interactions. In this review, we explore studies in Drosophila revealing that JAK-STAT signalling lies at the nexus of a complex network of interlinked pathways, including epidermal growth factor receptor (EGFR), c-Jun N-terminal kinase (JNK), Notch, Insulin, Hippo, bone morphogenetic protein (BMP), Hedgehog (Hh) and Wingless (Wg). These pathways can synergise with or antagonise one another to produce a variety of outcomes. Given the conserved nature of signal transduction pathways, we conclude with our perspective on the implication of JAK-STAT signalling dysregulation in human diseases, and how studies in Drosophila have the potential to inform and influence clinical research.

New insights into the neurophysiological effects of heat stress on the Chinese mitten crab (Eriocheir sinensis).

Climate warming and frequent incidents of extreme high temperatures are serious global concerns. Heat stress induced by high temperature has many adverse effects on animal physiology, especially in aquatic poikilotherms. Chinese mitten crab (Eriocheir sinensis) is sensitive to high temperatures, this study evaluated the harmful effects of heat stress on the neurotoxicity, intestinal health, microbial diversity, and metabolite profiles. The results showed that heat stress caused histopathological damages and altered the ultrastructure of lesions in the cranial ganglia. Heat stress significantly upregulated the mRNA expression of apoptosis-related genes, and significantly altered the expression of neurotransmitter receptors. In addition, heat stress induced significant intestinal damages that mainly manifested as a significant increase in the activity of diamine oxidase in the serum and contents of histamine in the intestine. The diversity and abundance of intestinal microbiota altered abnormally in E. sinensis exposed to heat stress, and the bacteria that exhibited significant variations in abundance were closely related to the production of neurotransmitters and neuromodulators. Heat stress caused significant changes in the intestinal metabolite profiles, which mainly involved the amino acid and lipid metabolism pathways. Analysis of the correlation showed that the abnormal changes in metabolites were closely related to differences in the abundance of intestinal microbiota. Therefore, this study showed that heat stress could cause neurophysiological toxic effects, which may be related to intestinal ecological imbalance.

Feed additives for methane mitigation: Recommendations for testing enteric methane-mitigating feed additives in ruminant studies.

There is a need for rigorous and scientifically-based testing standards for existing and new enteric methane mitigation technologies, including antimethanogenic feed additives (AMFA). The current review provides guidelines for conducting and analyzing data from experiments with ruminants intended to test the antimethanogenic and production effects of feed additives. Recommendations include study design and statistical analysis of the data, dietary effects, associative effect of AMFA with other mitigation strategies, appropriate methods for measuring methane emissions, production and physiological responses to AMFA, and their effects on animal health and product quality. Animal experiments should be planned based on clear hypotheses, and experimental designs must be chosen to best answer the scientific questions asked, with pre-experimental power analysis and robust post-experimental statistical analyses being important requisites. Long-term studies for evaluating AMFA are currently lacking and are highly needed. Experimental conditions should be representative of the production system of interest, so results and conclusions are applicable and practical. Methane-mitigating effects of AMFA may be combined with other mitigation strategies to explore additivity and synergism, as well as trade-offs, including relevant manure emissions, and these need to be studied in appropriately designed experiments. Methane emissions can be successfully measured, and efficacy of AMFA determined, using respiration chambers, the sulfur hexafluoride method, and the GreenFeed system. Other techniques, such as hood and face masks, can also be used in short-term studies, ensuring they do not significantly affect feed intake, feeding behavior, and animal production. For the success of an AMFA, it is critically important that representative animal production data are collected, analyzed, and reported. In addition, evaluating the effects of AMFA on nutrient digestibility, animal physiology, animal health and reproduction, product quality, and how AMFA interact with nutrient composition of the diet is necessary and should be conducted at various stages of the evaluation process. The authors emphasize that enteric methane mitigation claims should not be made until the efficacy of AMFA is confirmed in animal studies designed and conducted considering the guidelines provided herein.

Investigation of natural infection of BALB C mice by Bartonella henselae

Specific Pathogen-Free (SPF) animals are bred and maintained to exclude pathogens associated with significant morbidity or mortality, which may pose a risk to research replicability. The BALB/c strain is distributed globally and is among the most commonly used inbred strains in immunology and infectious disease research. Despite being a widely distributed bacterium that causes chronic infection, Bartonella henselae infection has not been investigated in any protocol that characterizes SPF animals. The objective of this study was to investigate the potential natural infection of laboratory animals of the BALB/c lineage by B. henselae. To achieve this, ten immunocompetent BALB/c mice were obtained directly from the bioterium and euthanized for collection of samples, including blood, skin, spleen, liver, heart, eye, kidney, intestine, esophagus, and brain. DNA was extracted using a commercial kit and tested via nested PCR for the ftsZ gene, as well as conventional PCR and qualitative real-time PCR using Sybr® Green for the citrate synthase gene (gltA), all specific reactions for B. henselae. All animals showed detection of B. henselae DNA in at least two different reactions in different tissues. The sequenced amplicons showed 100% similarity to B. henselae. The use of mice infected by B. henselae in experiments is undesirable, as the bacteria can affect several aspects of the animal's physiology and consequently influence the results of the project, especially when subjected to immunosuppression. More studies are needed to understand and confirm the natural infection in experimental animals by Bartonella spp. To date, no additional published reports of contamination of experimental animals by these bacteria have been identified.

Unique and Common Agonists Activate the Insect Juvenile Hormone Receptor and the Human AHR

Transcription factors of the bHLH-PAS family play vital roles in animal development, physiology, and disease. Two members of the family require binding of low-molecular weight ligands for their activity: the vertebrate aryl hydrocarbon receptor (AHR) and the insect juvenile hormone receptor (JHR). In the fly Drosophila melanogaster, the paralogous proteins GCE and MET constitute the ligand-binding component of JHR complexes. Whilst GCE/MET and AHR are phylogenetically heterologous, their mode of action is similar. JHR is targeted by several synthetic agonists that serve as insecticides disrupting the insect endocrine system. AHR is an important regulator of human endocrine homeostasis, and it responds to environmental pollutants and endocrine disruptors. Whether AHR signaling is affected by compounds that can activate JHR has not been reported. To address this question, we screened a chemical library of 50,000 compounds to identify 93 novel JHR agonists in a reporter system based on Drosophila cells. Of these compounds, 26% modulated AHR signaling in an analogous reporter assay in a human cell line, indicating a significant overlap in the agonist repertoires of the two receptors. To explore the structural features of agonist-dependent activation of JHR and AHR, we compared the ligand-binding cavities and their interactions with selective and common ligands of AHR and GCE. Molecular dynamics modeling revealed ligand-specific as well as conserved side chains within the respective cavities. Significance of predicted interactions was supported through site-directed mutagenesis. The results have indicated that synthetic insect juvenile hormone agonists might interfere with AHR signaling in human cells.

Trophic magnification rates of eighteen trace elements in freshwater food webs.

Trace elements play diverse roles in animal physiology ranging from essential micronutrients to potent toxicants. Despite animals accumulating many trace elements through their diets, relationships between trophic positions and biological concentrations of most trace elements remain poorly described. We report trophic transfer rates of Al, As, Ba, Cd, Co, Cu, Fe, Hg, Mn, Ni, Pb, Se, Sr, Ti, Tl, U, V, and Zn from 31 freshwaters located in distinct biogeographic regions. Elemental concentrations and stable nitrogen isotope ratios (proxies for trophic position) were determined in zooplankton, molluscs, insects, and fishes from all sites. Trophic magnification factors (TMFs) were calculated as the mean fraction of each element that transferred from prey to predators at each site. TMFs >1 indicate biomagnification and TMFs <1 indicate biodilution. Mercury was the only biomagnifying element (median TMF=3.77), and selenium neither biomagnified nor biodiluted (median TMF=1.01). All 16 remaining elements biodiluted, with median TMFs ranging from 0.07 (uranium) to 0.60 (thallium). We used a model selection procedure to determine whether intrinsic physical and chemical elemental properties explained differences in TMFs among elements. Elements with high covalent bonding indices (Q) had marginally greater TMFs than elements with low Q values. Based on their high Q values, we recommend investigation into the trophic transfer rates of ten additional trace elements, some of which may biomagnify through some aquatic food webs. The high variability in TMFs within elements suggests that ecological factors are likely more important than intrinsic elemental properties at determining elemental TMFs.

Analysis of Students' Science Process Skills in Practicum-Based Animal Physiology Learning

Research on student science process skills in practicum-based animal physiology learning with the aim of analysing the science process skills possessed by Biology Education students of FKIP Jambi University. The research method used is quantitative descriptive with data collection techniques in the form of observation sheets, interviews and documentation on the implementation of practicum activities carried out by 116 Biology Education Study Programme students. The results illustrate that in the implementation of animal physiology practicum activities, the science process skills of Biology Education students class of 2022 still have a less skilled category including classification skills at 51%, prediction at 52%, interpretation at 52% and applying concepts at 54% this is due to the lack of understanding of the concept of student theory taught by lecturers so that in practicum activities less mascimal in applying new theories, while the skills of planning experiments, using tools and materials, asking questions, observation, hypothesising, and communication are skilled categories. In conclusion, based on these results, the average student's science process skills are still categorised as less skilled, there are four skills with a percentage below 63% and an average skilled category of six skills with a percentage above 69%.

Bilirubin reductase shows host-specific associations in animal large intestines.

Animal gastrointestinal tracts contain diverse metabolites, including various host-derived compounds that gut-associated microbes interact with. Here, we explore the diversity and evolution of bilirubin reductase, a bacterial enzyme that metabolizes the host-derived tetrapyrrole bilirubin, performing a key role in the animal heme degradation pathway. Through an analysis of the bilirubin reductase phylogeny and predicted structures, we found that the enzyme family can be divided into three distinct clades with different structural features. Using these clade definitions, we analyzed metagenomic sequencing data from multiple animal species, finding that bilirubin reductase is significantly enriched in the large intestines of animals and that the clades exhibit differences in distribution among animals. Combined with phylogenetic signal analysis, we find that the bilirubin reductase clades exhibit significant associations with specific animals and animal physiological traits like gastrointestinal anatomy and diet. These patterns demonstrate that bilirubin reductase is specifically adapted to the anoxic lower gut environment of animals and that its evolutionary history is complex, involving adaptation to a diverse collection of animals harboring bilirubin-reducing microbes. The findings suggest that bilirubin reductase evolution has been shaped by the host environment, providing a new perspective on heme metabolism in animals and highlighting the importance of the microbiome in animal physiology and evolution.

Central mechanisms of muscle tone regulation: implications for pain and performance

Muscle tone represents a foundational property of the motor system with the potential to impact musculoskeletal pain and motor performance. Muscle tone is involuntary, dynamically adaptive, interconnected across the body, sensitive to postural demands, and distinct from voluntary control. Research has historically focused on pathological tone, peripheral regulation, and contributions from passive tissues, without consideration of the neural regulation of active tone and its consequences, particularly for neurologically healthy individuals. Indeed, simplistic models based on the stretch reflex, which neglect the central regulation of tone, are still perpetuated today. Recent advances regarding tone are dispersed across different literatures, including animal physiology, pain science, motor control, neurology, and child development. This paper brings together diverse areas of research to construct a conceptual model of the neuroscience underlying active muscle tone. It highlights how multiple tonic drive networks tune the excitability of complex spinal feedback circuits in concert with various sources of sensory feedback and in relation to postural demands, gravity, and arousal levels. The paper also reveals how tonic muscle activity and excitability are disrupted in people with musculoskeletal pain and how tone disorders can lead to marked pain and motor impairment. The paper presents evidence that integrative somatic methods address the central regulation of tone and discusses potential mechanisms and implications for tone rehabilitation to improve pain and performance.

Supporting rotational grazing systems with virtual fencing: paddock transitions, beef heifer performance, and stress response.

Animal welfare is integral to sustainable livestock production, and pasture access for cattle is known to enhance welfare. Despite positive welfare impacts, high labour requirements hinder the adoption of sustainable grazing practices such as rotational stocking management. Virtual fencing (VF) is an innovative technology for simplified, less laborious grazing management and remote animal monitoring, potentially facilitating the expansion of sustainable livestock production. VF uses Global Navigation Satellite System technology, wireless communication, and stimuli (auditory and electrical) to manage livestock movements and contain animals without physical barriers. Training animals to associate the auditory cue with the subsequent aversive stimulus enables effective livestock containment without physical barriers. While previous studies have largely dispelled concerns about adverse effects on cattle behaviour associated with the use of VF collars, there is limited knowledge regarding the impacts on animal physiology, particularly in rotational stocking systems. Addressing this knowledge gap, this study investigated differences in diet digestibility, livestock performance, and stress response of beef heifers on pastures using a VF compared to a physical electric fence. The study was conducted over 8weeks, subdivided into two grazing cycles, with 32 heifers in four groups. Each experimental pasture was subdivided into four paddocks. The study monitored the interaction with the VF by analysing the temporal development of the ratio of auditory and electrical cues (success ratio and confidence ratio) emitted by the collars. Additionally, the grassland herbage quality, BW gain, and concentrations of faecal cortisol metabolites (FCMs) were assessed, as well as the time required for animals to cross into a new paddock. VF success ratios increased in the second grazing cycle, reflecting enhanced adaptation over time. Similarly, the reduction in time taken to cross into new paddocks in the VF groups indicated that animals learned to interact with the VF and rely on the auditory cues for directing movements. The absence of a significant effect of the fencing system on FCMs suggested that stress was unrelated to the VF technology. Further, animal performance was not affected as indicated by similar BW gains under both fencing systems. This study also attempts to establish a benchmark threshold for successful responses to the auditory cues, allowing comparative evaluation of VF systems. Overall, under rotational grazing, VF did not adversely impact animal welfare or performance compared to physical fencing, opening avenues for further exploration of VF technology in diverse grazing conditions.

Predicting population-level impacts of projected climate heating on a temperate freshwater fish.

Climate heating has the potential to drive changes in ecosystems at multiple levels of biological organization. Temperature directly affects the inherent physiology of plants and animals, resulting in changes in rates of photosynthesis and respiration, and trophic interactions. Predicting temperature-dependent changes in physiological and trophic processes, however, is challenging because environmental conditions and ecosystem structure vary across biogeographical regions of the globe. To realistically predict the effects of projected climate heating on wildlife populations, mechanistic tools are required to incorporate the inherent physiological effects of temperature changes, as well as the associated effects on food availability within and across comparable ecosystems. Here we applied an agent-based bioenergetics model to explore the combined effects of projected temperature increases for 2100 (1.4, 2.7, and 4.4°C), and associated changes in prey availability, on three-spined stickleback (Gasterosteus aculeatus) populations representing latitudes 50, 55, and 60°N. Our results showed a decline in population density after a simulated 1.4°C temperature increase at 50°N. In all other modeled scenarios there was an increase (inflation) in population density and biomass (per unit area) with climate heating, and this inflation increased with increasing latitude. We conclude that agent-based bioenergetics models are valuable tools in discerning the impacts of climate change on wild fish populations, which play important roles in aquatic food webs.

The role of NMDA receptors in fish stress response: Assessments based on physiology of the caudal neurosecretory system and defensive behavior.

Stress strongly influences the physiology and behavior of animals, and leads into a pathological condition and disease. NMDA receptors (NMDARs) play a crucial role in the modulation of neural activity. To understand the role of NMDARs in fish stress response, we used NMDARs agonist aspartate to test the functional role of its input on the Dahlgren cell population in the caudal neurosecretory system (CNSS) of the olive flounder. In addition, the effect of the NMDARs antagonist D-AP5 on the expression of genes of the main secretory products of the CNSS after stress was investigated by using qPCR technology and the effect of the NMDARs antagonist D-AP5 on post-stress behavior was explored by behavioral methods. Ex vivo electrophysiological experiments showed that the NMDARs agonist aspartate enhanced the firing frequency of Dahlgren cells. Additionally, aspartate treatment increased the incidence of cells exhibiting bursting firing pattern, this result is corroborated by the observed upregulation in the expression of ion channels and major hormone genes in the CNSS. Furthermore, the excitatory influence of aspartate was effectively counteracted by NMDARs antagonist D-AP5. Interestingly, NMDARs antagonist D-AP5 treatment also significantly decreased the plasma cortisol levels and the expression of CRH, UI, and UII in CNSS after acute stress. Treatment with D-AP5 effectively attenuated the stress response, as evidenced by alterations in respiratory metabolism, sand-burying behavior, swimming distance, simulated capture, and escape response. In conclusion, modulation of Dahlgren cell excitability in the CNSS by NMDARs contributes to the regulation of the stress response, NMDARs antagonist D-AP5 can effectively suppress stress response in flounder by regulating the stress hormone expression and secretion. CLINICAL TRIAL REGISTRATION: Project code SHOU-DW-2022-032.

Acoustic-based models to assess herd-level calves' emotional state: A machine learning approach

Animal bioacoustics is an important tool for monitoring different aspects of the physiology, behavior and well-being of animals remotely, non-invasively and continuously. Studies using this science are growing mainly due to the development of machine learning. This work aims to investigate the use of machine-learning classifiers to determine whether calves’ vocalization audio data can be used to assess their welfare condition regarding feeding. Firstly, we collected several calves’ vocalization audio data before and after feeding the animals at different day times and ages. Then, eleven time-domain, frequency-domain and sound quality-based metrics were extracted from these audio data and used as features for the classifiers. These features were used to determine whether vocalization audio data belonged to before or after feeding classes. Moreover, the most relevant ones were identified using the Random Forest algorithm. Finally, seven machine-learning classifiers were trained and tested, considering the entire set of features and a subset containing the most relevant features. The k-nearest neighbor classifier trained with the subset of the most relevant features obtained a 98.37% accuracy. Both frequency-domain and sound quality features played important roles in this classification. The main implications of this study are the development of a methodological proposal to study acoustics using machine learning and the fact that vocalization is a biomarker of animal welfare.

Editorial: Diving deeper with metabolomics into animal physiology

Editorial: Diving deeper with metabolomics into animal physiology

Authentic-Inquiry-Project about Nyale on Science Teachers Critical and Creative Thinking Skills

The aim of this study was to determine the effect of authentic inquiry project practicum on Nyale in improving critical and creative thinking skills of prospective science teacher students. The method used was experimental research method with one group pretest postest. The research subjects involved science education students of one of the universities in Sumba in the 2023/2024 academic year who contracted animal anatomy and physiology courses. The instrument used in this research was a critical and creative thinking skills test. The results of the Wilcoxon test showed that in experimental classes 1 and 2 there was an effect of practicum on the critical and creative thinking skills of prospective science teacher students as evidenced by the Asymp. Sig. (2-tailed) 0.000 &lt; than 0.05. In addition, the N-gain test results showed an increase in critical thinking skills before and after practicum in experimental classes 1 and 2 with moderate categories. While the N-gain test of creative thinking skills showed an increase in the creative thinking skills of prospective science teachers in the moderate category for experimental class 1 and in the low category for experimental class 2. From the results obtained, it can be concluded that the authentic inquiry project practicum on Nyale is effective in improving the critical and creative thinking skills of prospective science teachers.