Changes to Australia's climate continue to be increasingly relevant for human, animal and planetary health, and have far-reaching impacts on disease emergence and food security. There is a deficit of knowledge in Australia around how climatic conditions are affecting infectious animal diseases, beyond the more obvious spatial distribution changes of parasites and disease vectors. It is critically important for veterinarians and stakeholders to better understand how animal diseases will change with a changing climate. Disease predictions for our companion animals, livestock species and wildlife will allow for better patient care, prophylaxis, planning and disease mitigation. This scoping review sought to survey the literature about the impact of climate on non-vector-borne infectious diseases in Australia for all vertebrate animal species. An electronic search was conducted in four databases. A three-level screening process was conducted on the 489 records captured by two independent reviewers to yield 28 eligible primary research articles published between 1983 and 2023. Frogs and their fungal disease, chytridiomycosis, were the most frequent species and disease studied. Rainfall, temperature and season were the most frequent climatic variables studied. Eighteen out of 21 eligible articles found a statistically significant association between an animal disease and a climatic variable. Of these, eight articles found an association with temperature, while seven articles found an association each with rainfall and season. Study designs were predominantly either cross-sectional or longitudinal. Longitudinal study designs are critical for producing strong evidence of climatic associations and Australia would benefit from more studies of this kind. Overall, there is a need for more research into how climate will continue to drive non-vector-borne animal disease trends in Australia, particularly for our companion animals and our production species.

Ethanol is a neuroactive compound known to alter cognition, behavior, and physiology in a diverse range of vertebrate and invertebrate animals. It is also naturally present in many environments and might exert influence on various organisms, such as honeybees feeding on fermenting nectar, reflecting on the ecological networks the organism is involved in. In this study, we tested whether exposure of honeybees to low ethanol concentrations (1%) induces 'pessimistic' or 'optimistic' cognitive judgment bias (CJB), an information processing pattern thought to be related to affective processes. Bees were trained in a classical olfactory conditioning paradigm and then presented with conditioned stimuli and ambiguous odor mixtures to assess changes in decision-making under uncertainty. We additionally measured locomotor performance and ethanol absorption via spectrophotometric analysis of hemolymph. Ethanol levels peaked rapidly and remained elevated throughout the testing period. Despite this, ethanol had no detectable effect on choices in the CJB assay or locomotor behavior. These findings suggest that acute exposure to low-dose ethanol does not induce affect-like changes in honeybees. Our study represents the first test of pharmacologically induced judgment bias in an invertebrate and contributes to understanding the resilience of insect cognition to natural neuroactive compounds. The results highlight the limits and potential of CJB paradigms for probing affect-like states across taxa and support the honeybee as a valuable model in comparative cognition and the behavioral ecology of neuroactive substance exposure.

Ricin is considered a chemical and biological weapon. It is found in the seeds of a plant, Ricinus communis. There are different isoforms of ricin with different levels of toxicity, depending on the R. communis plant. There is no current prophylaxis and specific treatment for ricin poisoning is recent. Assessing potential countermeasures against this toxin still relies on testing on vertebrate animal models. Galleria mellonella larva, already used for the testing of bacteria, viruses, and fungi, represents an alternative model that is more ethical, inexpensive, and easier to use. In this study, we demonstrated for the first time the sensitivity of G. mellonella larvae to different cultivars of ricin. We observed mortality and a reduction in health index scoring over the days of testing. The health index scoring was based on the survival, the melanization, the mobility, and the capacity of larvae to produce a cocoon or not. Mortality was cultivar- and dose-dependent. Mortality of G. mellonella larvae was reduced when they were treated with a monoclonal antibody and concomitantly injected with ricin. Thus, G. mellonella represents a rapid and simple model of ricin poisoning, and, more particularly, a relevant model to test new therapeutics against ricin.

Physiology textbooks consistently describe how the repeating sarcomeric organization of vertebrate skeletal and cardiac muscle generates a striated appearance. Unfortunately, very few texts discuss the common functional role of this sarcomeric organization. Briefly, the repeating sarcomeres in striated muscles provide a mechanism to amplify the nanometer-scale steps of myosin cross bridges into the dynamic large-scale movements embodied by diverse animals; that is, the sarcomeres contracting in unison dramatically expand the speed of muscle shortening. This functional role was recognized by the proponents of the sliding filament theory in the mid-1950s. Striated muscles are found not only in vertebrate skeletal and cardiac muscles but are also used to power movement in animals as disparate as jellyfish, clams, swimming crabs, and flying insects. Historically, several different invertebrate muscles have been studied in great detail. Recent studies based on whole sequenced genomes and transcriptome sequencing are providing new evidence that striated muscles evolved at the dawn of animal life. Although there is a deep evolutionary divergence of striated and smooth muscle myosins, striated muscles have likely arisen multiple times through convergent evolution. Several well-studied examples of these striated muscles from diverse animals are discussed to illustrate their common functional roles. The central thesis presented here is that striations provide a structural mechanism to amplify the speed of muscle shortening. Comprehending this principle is foundational to the teaching of basic muscle structure and function relationships.NEW & NOTEWORTHY Although physiology educators recognize that skeletal and cardiac muscles are striated, few physiology texts explain the functional role of these striations. The repeated sarcomeres that form striations in these muscles provide a mechanism to amplify the speed of shortening. The striated muscles observed in vertebrate animals are but one example of a more general pattern. The current paper explores the evolutionary origins of striated muscles and presents examples of these muscles from diverse animals.

The rhizosphere, a critical soil layer around plant roots, is enriched with carbon from root exudates, influencing microbial communities that can either protect against or cause plant diseases. Bacteriophages significantly impact soil nutrient cycles and ecosystem processes through cell lysis and horizontal gene transfer. They play a vital role in the rhizosphere by affecting plant stress responses and climate adaptation. Bacteriophages exert a range of negative effects on Actinobacteria, impacting their ecological and physiological functions by diminishing Actinobacteria's roles in antibiotic production, soil health, and plant growth. Phage predation affects nutrient cycling by influencing nitrogen and carbon metabolism, with evidence showing that phages can alter microbial diversity and function, leading to changes in soil ammonium levels and carbon decomposition rates. In wastewater treatment, bacteriophages can improve process efficiency by targeting harmful bacteria, managing foam formation, and enhancing sludge reduction through enzymatic action. Additionally, bacteriophage dispersal mechanisms in the rhizosphere can be enhanced by rhizosphere-associated animals. Numerous invertebrate and vertebrate animals can significantly alter the rhizosphere environment by amplifying, mobilizing, and distributing both phages and bacterial hosts. Herein, three main mechanisms by which animals enhance the dispersal of bacteriophages in the rhizosphere are discussed. This review discusses bacteriophages' roles in soil ecosystems, highlighting their impact on nutrient cycling, plant health, and soil remediation, as well as animal-mediated phage dispersal mechanisms. Overall, while bacteriophages have potential biotechnological applications, their negative effects on microbial functions and nutrient cycling highlight the need for balanced use and further research.

Delineating the genetic and environmental instigators of congenital heart disease (CHD), affecting up to 1% of newborns, will improve preventative, diagnostic, and therapeutic efforts to mitigate disease outcomes. Although mutations in NAA15, which encodes the auxiliary subunit of N-alpha-acetyltransferase A (NatA), are associated with CHD in humans, vertebrate animal models of NAA15-related heart disease have yet to be described. Therefore, we isolated zebrafish strains carrying null mutations in paralogs naa15a and naa15b, which we co-localized to the early larval myocardium. Double knockout (DKO) naa15-deficient larvae exhibited diminutive, lowly contractile, and bradycardic ventricles composed of fewer and smaller cardiomyocytes (CMs) incapable of proliferation. On a subcellular level, mutant CMs exhibited moderately disorganized myofibrils. CM-specific re-expression of naa15a partially rescued the contractility and growth deficits, revealing an indispensable myocardial function. Ubiquitous mis-expression of human NAA15 achieved complete rescue, enabling functional testing of human NAA15 variants. Animals with a reduced dosage (RD) of naa15 survived to adulthood and shared phenotypes with DKO larvae, including smaller ventricular chambers and CMs, which exhibited mosaic myofibril disarray. Deep quantitative proteomic profiling of WT and naa15 RD adult hearts revealed differential expression of multiple protein classes, including several subunits of mitochondrial respiratory complex I, a component of the electron transport chain. Accordingly, we documented reduced mitochondrial content and function in the myocardium of naa15-deficient larvae. Taken together, our data reveal myocardial functional and structural abnormalities associated with mitochondrial dysfunction in a vertebrate animal model of NAA15-related heart disease.

This research was motivated by the need for science learning on animal classification materials that can improve conceptual understanding, creativity, and collaboration among elementary school students. The purpose of this study was to determine the effect of implementing project-based learning with a STEAM approach through educational games on the learning outcomes of third-grade students at SDN 42 Pekanbaru. This study employed a quantitative approach with a quasi-experimental method using a one-group pretest-posttest design involving 33 students. The implementation was conducted in two meetings, consisting of pretest administration, material delivery using the Cambridge module, group formation, implementation of an educational game project entitled Discover Animal Classification, and posttest administration. Data were collected through learning outcome tests and analyzed descriptively and inferentially. The results showed that project-based learning with a STEAM approach through educational games was able to improve students’ understanding in distinguishing vertebrate and invertebrate animals, classifying animals based on their diets, and identifying their locomotor organs. In addition, students became more active, enthusiastic, creative, and able to collaborate effectively in groups. Therefore, this learning model is effective in improving learning outcomes and developing 21st-century skills among elementary school students.

Enterocytozoon bieneusi is a widespread endoparasite, commonly found in the gastrointestinal tract of humans and various vertebrate and invertebrate animals. It is the most common cause of microsporidiosis in humans, particularly among immunosuppressed individuals. Given the limited epidemiological data in Portugal, this study aimed to estimate the occurrence of E. bieneusi, perform genetic characterization, and explore its circulation dynamics in small mammal populations of Trás-os-Montes region. A total of 294 faecal samples from rodents and shrews non-invasively collected were analysed using a nested polymerase chain reaction (PCR) assay targeting the internal transcribed spacer region and flanking ribosomal RNA coding sequences of E. bieneusi. The parasite DNA was detected in two rodent samples: wood mouse (Apodemus sylvaticus) and Lusitanian pine vole (Microtus lusitanicus), corresponding to an overall occurrence of 0.68%. Our findings represent the first report of E. bieneusi in Lusitanian pine vole and the first detection of this parasite in small mammals from Portugal. Both samples belonged to the zoonotic Group 1, with one identified as genotype C. These findings provide baseline data on the distribution and host range of E. bieneusi in Portuguese wildlife and may guide future surveillance efforts within a One Health framework.

Bimodal gene expression systems have played a major role in uncovering the function of genes, cells and organ systems during and after development. Employed initially in model systems such as flies and mice, advances in gene technology have vastly expanded the number of species in which these systems can be deployed. One of their limitations is the challenge of imposing temporal expression control. Here, we report the incorporation of temperature-sensitive intein modules with different temperature profiles into QF2, the transcription factor anchoring the Q-system widely used in Drosophila and introduced recently into several insect species and the zebrafish D. rerio. Intein removal from temperature-sensitive QF2_INTts activators in Drosophila larvae and adult flies raised under permissive conditions (18°C to 25°C) renders QF2 active and drives strong expression of QUAS reporters. In contrast, raising Drosophila at restrictive temperatures (23°C to 30°C) keeps QF2_INTts in a non-functional state unable to bind DNA and therefore, keeping QUAS reporters inactive. We further show that reporter expression can be turned on and off both during development and in flies by changing temperature conditions between restrictive to permissive. Finally, QF2_INTts animals carrying QUAS-Kir2.1 encoding the inward rectifying potassium channel raised under restrictive conditions are fully viable, while raised at or moved as adults to permissive temperature results in embryonic lethality or leads to paralysis. Thus, the intein-based Q-system renders the difficult to employ drug-dependent suppressor QS unnecessary, greatly facilitating temporal regulation of gene expression. Having several advantages over the GAL4 system, the Q system has gained broad acceptance in other insect species and the zebrafish D. rerio, and thus QF2_INTts drivers can be implemented in many other organisms, including disease-transmitting mosquitoes and poikilotherm vertebrate animals much more closely related to humans, to study gene and cell function at any time during or after development.

This article analyzes the effectiveness of integrating modern pedagogical technologies into the teaching of Zoology, specifically the "Vertebrate Animals" section, within higher education frameworks. In contrast to traditional rote learning, this study highlights the role of 3D modeling, virtual laboratories, and Problem-Based Learning (PBL) in reinforcing student knowledge. Research results demonstrate that an innovative approach not only enhances theoretical comprehension but also develops practical analytical skills and creative thinking.

Introduction. Chronic stress and paracetamol exposure are among the most relevant factors affecting neurochemical and molecular processes in the brain in modern humans. Excessive activation of inflammatory and oxidative pathways during chronic psychosocial stress can lead to a decrease in the expression of neuroprotective and antioxidant genes, exacerbating the neurotoxic effects of drugs such as paracetamol.Objective. To assess the effect of chronic unpredictable stress and oral administration of paracetamol, as well as their combined effect on the morphological state of the brain and on the expression of genes of the antioxidant system and neurotrophic factor in the hippocampus in rats.Materials and methods. The experiment was conducted on forty eight rats (males and females in equal numbers), divided into four groups: control, chronic stress, paracetamol, combined exposure to stress and paracetamol. A complex of various stressors was used to model chronic stress. Paracetamol was administered orally at a dose of 1000 mg/kg. The morphological state of the hippocampus was assessed histologically, gene expression - by quantitative real-time PCR. Statistical analysis included bootstrapping and correction for multiple comparisons using the Holm-Bonferroni method.Results. Macroscopic and histological examination of the rat brain showed the preservation of the normal anatomical structure and integrity of the hippocampal tissues in all groups. Statistical analysis of gene expression showed animals treated with paracetamol (groups III and IV) to show a significant decrease in the mRNA levels in the Sod1 and Bdnf genes in the hippocampus compared to the control and the group exposed to stress only (group II). The expression level of Sod1 and Bdnf was significantly lower in both males and females in these groups (p = 0.001). For the Hmox1 gene, a significant decrease in expression was found only in males in the paracetamol-treated group, while no changes were noted in females.Limitations. Laboratory animals of the same biological species were used for the experiment, and the toxicant was used in only one concentration.Conclusion. Thus, paracetamol, both in isolated use and in combination with chronic stress, has a more pronounced inhibitory effect on the expression of key genes of antioxidant protection and neurotrophic support than stress alone.Compliance with ethical standards. Date of the meeting of the bioethics commission of theUfa Research Institute of Occupational Medicine and Human Ecology dated 02/08/2024 No. 01–02. Throughout the study, the animals were kept in standard conditions with 12-hour artificial lighting during the daytime, a relatively constant level of humidity (30–70%) and an air temperature of 20–25 °C. All animal manipulations were carried out in strict compliance with the rules prescribed in the basic regulatory documents, including the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (Strasbourg, 1986) and the Helsinki Declaration on the Humane Treatment of Animals.Contributions: Gizatullina A.A. – concept and study design, material collection and processing, statistical analysis, manuscript writing; Valova Ya.V. – material collection and processing, statistical analysis; Mukhamadieva G.F. – editing; Karimov D.O. – concept and study design, statistical analysis; Karimov D.D. – material collection and processing; Ryabova Yu.V. – material collection and processing; Khusnutdinova N.Yu. – material collection and processing; Khmel A.O. – material collection and processing. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.Conflict of interest. The authors declare no conflict of interest.Funding. Industry research program of Rospotrebnadzor for 2021–2025 “Scientific substantiation of the national system for ensuring sanitary and epidemiological welfare, managing health risks and improving the quality of life of the population of Russia”, on the topic: 6.9.1.2 “Study of the impact of chemical production factors under conditions of chronic stress” No. NIOKTR I124021200153-3.Received: April 30, 2025 / Revised: July 9, 2025 / Accepted: October 15, 2025 / Published: April 17, 2026

Prototheca are unicellular, non-pigmented, yeast-like microalgae widely distributed in natural and anthropogenic environments. While typically saprophytic, these organisms are the only plant species known to have emerged as opportunistic pathogens for vertebrate animals including humans. The algae have been recognised as ubiquitous inhabitants of domestic and municipal sewage, considered as point sources of Prototheca contamination in natural aquatic systems. These observations, however, have been based on data from a very limited number of historical studies. The purpose of this work was to readdress the question of wastewater as a habitat for Prototheca, using a new and refined experimental methodology, and in light of the recently updated taxonomy and nomenclature of the Prototheca genus. Over a three-month period, 96 samples were collected from three municipal wastewater treatment plants, at four different stages of the treatment process. The algae were recovered from 74 samples, which translates into an overall prevalence of 78.1%. The highest Prototheca population was found in raw sewage, and decreased progressively with each purification stage. The Prototheca cell count in the effluent was 100-fold lower than in the untreated sewage, resulting in an algal removal efficiency of up to 99.9%. The occurrence of Prototheca in treated wastewater suggests that wastewater treatment plants may act as a pathway for the release of these algae into receiving natural environments. In both influent and effluent, species distribution was dominated by Prototheca species previously reported as pathogenic, with P. wickerhamii detected most frequently. The study concludes by advocating for regular monitoring of pre- and post-treatment wastewater for Prototheca species and emphasising the need for guideline limits on their acceptable levels at both treatment stages.

Larvae of the greater wax moth Galleria mellonella have become an increasingly important in vivo model for biomedical research, providing a practical, ethical, and biologically relevant alternative to vertebrate organisms. Its suitability as a model system lies in its low maintenance cost, ease of handling, and ability to survive at both ambient and mammalian body temperatures. Most importantly, G. mellonella exhibits an innate immune system with functional and structural parallels to that of mammals, allowing meaningful insights into infection dynamics and immune responses. This review summarizes the biological and immunological foundations that underpin the use of G. mellonella in experimental research and examines its expanding range of applications. The model has been successfully employed to study microbial pathogenicity, antimicrobial efficacy, host-pathogen interactions, and toxicological responses. In recent years, its use has extended to emerging fields such as nanomedicine, immunomodulation, and environmental biotechnology, reflecting its growing translational value. The adoption of G. mellonella also aligns with current ethical principles in science, particularly the 3Rs framework (replacement, refinement, and reduction), by minimizing the use of vertebrate animals while maintaining robust experimental outcomes. However, certain challenges persist, including the lack of adaptive immunity and the need for methodological standardization to enhance data reproducibility and comparability across laboratories. Collectively, the growing body of evidence supports G. mellonella as a reliable and versatile experimental model that bridges the gap between invertebrate and mammalian systems. Continued methodological refinement and integration with molecular and omics approaches are expected to further consolidate its role in translational and preclinical research. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC.

Potentials of newly selected chicken genotypes for dual-purpose under organic conditions.

Dopamine is one of the best-known neurotransmitters found in most animals. Among the major roles that this mediator plays in many organisms is the regulation of motor skills, behavior, and feeding. Its localization in the central (CNS) and peripheral nervous systems (PNS) has been well studied mainly in vertebrate animals, but remains poorly understood in invertebrates such as mollusks of the class Bivalvia. Nevertheless, bivalves are of particular interest for their nervous system that has undergone a number of simplifications due to the sedentary lifestyle. As shown in the review, the key functions of dopamine have been retained in this group, with, however, a shift towards regulation of effector organs and physiological processes. The interaction between serotonin and dopamine, which regulates the degree of motor activity, nutrition, and locomotion, also deserves special consideration. There still remains a vast number of unresolved issues concerning the effects that dopamine exerts in the bivalve CNS, its role in the regulation of larval development and behavior of adults. This review summarizes the major known aspects of dopamine, including its localization and role in the life history of bivalves.

Primary Ciliary Dyskinesia (PCD) is a genetically heterogeneous disorder leading to destructive airway disease with severe bronchiectasis and chronic lung failure in adulthood. Pathogenic variants in CCDC40 are associated with more severe reduction of lung function compared to most other PCD types. Currently, no therapies correcting the underlying disease mechanism are available. Here we investigate the efficacy of lipidoid nanoparticle-formulated mRNA encoding human CCDC40 (LNP-CCDC40-mRNA) as a corrective measure for structural and functional defects in vitro (human cells) and in vivo (zebrafish). Human nasal respiratory epithelial cells cultured at air-liquid-interface from five CCDC40-deficient individuals and a newly generated vertebrate animal model (ccdc40-/- zebrafish) were treated with LNP-CCDC40-mRNA. CCDC40-deficient cells were analyzed by high-speed video microscopy and immunofluorescence microscopy. ccdc40-/- zebrafish olfactory pit cilia were analyzed by high-speed video microscopy and fluid flow assays. Topical application of exogenous LNP-CCDC40-mRNA to CCDC40-deficient cells results in endogenous CCDC40 expression (10-74% of ciliated cells), enabling axonemal integration of CCDC40-associated proteins (CCDC39, GAS8/DRC4, DNALI1). Consistently, ciliary beat frequencies were significantly increased in treated CCDC40-deficient cells and comparable to healthy control cells. Further, we showed improved ciliary transport of fluorescent particles. Injection or topical application of human LNP-CCDC40-mRNA to ccdc40-/- zebrafish significantly increased ciliary motility and established directional flow in olfactory pits. We provide structural and functional evidence in vitro and in vivo for the biological efficacy of LNP-CCDC40-mRNA in CCDC40-deficient respiratory cells and zebrafish. Based on our results, an in vivo human study (Phase 1 trial) is planned in individuals with pathogenic variants in CCDC40.

Mapping behavior of individual vertebrate animals across lifespan could provide an unprecedented view into the lifelong process of aging. We created a platform for high-resolution continuous behavioral tracking of the African killifish across natural lifespan from adolescence to death. We found that animals follow distinct individual aging trajectories. The behaviors of long-lived animals differed markedly from those of short-lived animals, even relatively early in life, and were linked to organ-specific transcriptomic shifts. Machine-learning models accurately inferred age and even forecasted an individual's future lifespan, given only behavior at a young age. Finally, we found that animals progressed through adulthood in a sequence of stable and stereotyped behavioral stages with abrupt transitions, revealing precise structure for an architecture of aging.

Chloride contamination in drinking water resulting from natural sources, industrial effluents, and disinfection processes, poses significant risks to aquatic life, particularly during early developmental stages making the evaluation of its ecological impact essential. Although chloride is essential in small amounts, elevated concentrations can adversely affect survival, growth, and physiological processes in aquatic organisms. This study aimed to determine the threshold toxicity of chloride in water and assess acute lethal effects on zebrafish (Danio rerio) embryos. Fertilized embryos at 2 hours post-fertilization (hpf) were exposed to five chloride concentrations (2,4,6,8 and, 10 mgL-1), with tap water and distilled water serving as controls. Twenty embryos per each and duplicated. Toxicological endpoints including mortality, hatching rate, heart rate, and morphological abnormalities, were assessed until 96 hpf. The results demonstrate a clear dose-dependent increase in mortality, with the highest rate of 68.75±0.58% was observed at 10 mgL-1and the lowest rate of 25±0.58% was observed at 2 mgL-1. The hatching rate declined gradually at higher concentrations, showing 50±0.00% at 2 mgL-1, and further decreased to 12.5±1.2% at 10 mgL-1, and heart rate was increased progressively, reaching 192±2 bpm at 10 mgL-1compared to 140±4 bpm in controls. Embryos in tap water exhibited 12.5±0.58 % mortality, 75±1.74% hatching, and a heart rate of 150±2 bpm at 96 hpf. Morphological abnormalities, including spinal curvature at 2 mgL-1 and yolk sac oedema at 8 mgL-1, were observed with an occurrence rate of 20% of the larvae. The calculated LC50 value of chloride is 5.6 mgL-1. These findings emphasize the need to reconsider water quality guidelines to protect vulnerable early developmental stages of aquatic life. Overall, this research demonstrates that even relatively low chloride concentrations can induce significant developmental and morphological effects in zebrafish embryos, supporting their application in environmental risk assessment and guiding safer drinking water management. Keywords: Chloride, Hatching rate, Morphological abnormalities, Mortality rate, Zebrafish

Botulinum neurotoxins: A distinct subclass of SNARE-Cleaving endopeptidases.

The transmission of bacterial, viral, fungal, or parasitic agents from vertebrate animals to humans constitutes a group of diseases called zoonoses. The frequency of these zoonoses is increasing, and new zoonoses are emerging due to travel, factory farming, biodiversity loss, climate change, and deforestation. These diseases have a significant impact on public health, both human and animal, as well as on food, environmental management and the economy. This second article reviews the viral, fungal, and parasitic zoonoses with cutaneous manifestations, known as dermato-zoonoses.