Animal Systems Research Articles

Genetic variation in the brooding brittle-star: a global hybrid polyploid complex?

The widespread and abundant brooding brittle-star (Amphipholis squamata) is a simultaneous hermaphrodite with a complex mitochondrial phylogeography of multiple divergent overlapping mtDNA lineages, high levels of inbreeding or clonality and unusual sperm morphology. We use exon-capture and transcriptome data to show that the nuclear genome comprises multiple (greater than 3) divergent (π > 6%) expressed components occurring across samples characterized by highly divergent (greater than 20%) mitochondrial lineages, and encompassing several other genera, including diploid dioecious species. We report a massive sperm genome size in A. squamata, an order of magnitude larger than that present in other brittle-stars, and consistent with our SNP-based measure of greatly elevated ploidy. Similarity of these genetic signatures to well-known animal systems suggests that A. squamata (and related taxa) is a hybrid polyploid asexual complex of variable subgenome origins, ploidy and reproductive mode. We discuss enigmatic aspects of A. squamata biology in this light. This putative allopolyploid complex would be the first to be reported from the phylum Echinodermata.

Prospects of Electrocorticography in Neuropharmacological Studies in Small Laboratory Animals.

Electrophysiological methods of research are widely used in neurobiology. To assess the bioelectrical activity of the brain in small laboratory animals, electrocorticography (ECoG) is most often used, which allows the recording of signals directly from the cerebral cortex. To date, a number of methodological approaches to the manufacture and implantation of ECoG electrodes have been proposed, the complexity of which is determined by experimental tasks and logistical capabilities. Existing methods for analyzing bioelectrical signals are used to assess the functional state of the nervous system in test animals, as well as to identify correlates of pathological changes or pharmacological effects. The review presents current areas of applications of ECoG in neuropharmacological studies in small laboratory animals. Traditionally, this method is actively used to study the antiepileptic activity of new molecules. However, the possibility of using ECoG to assess the neuroprotective activity of drugs in models of traumatic, vascular, metabolic, or neurodegenerative CNS damage remains clearly underestimated. Despite the fact that ECoG has a number of disadvantages and methodological difficulties, the recorded data can be a useful addition to traditional molecular and behavioral research methods. An analysis of the works in recent years indicates a growing interest in the method as a tool for assessing the pharmacological activity of psychoactive drugs, especially in combination with classification and prediction algorithms.

Treatment of functional disorders of gastrointestinal tract of calves with complex phytomineral preparation

Significant economic losses due to diarrhoea morbidity in newborn young animals lead to a decrease in the profitability of the sector in the structure of agricultural production. Therefore, the purpose of this study was to create and investigate the therapeutic effect of application of complex phytomineral preparation using local components: opoka of Taskala deposit and decoction of camelthorn (Alhagi maurorum) for treatment of diarrhoea in calves. The therapeutic effect of the preparation was studied in the conditions of dairy farm in Kyzylorda region on 5-7-day-old calves. The effect of the preparation was studied in a comparative aspect with an analogous preparation of imported production and the results obtained from healthy animals. Clinical, haematological, and biochemical methods of investigation were employed. The findings obtained suggest a high therapeutic effect of the created preparation. Its application allowed reducing the duration of the treatment period by 5-7 days compared to the imported preparation and restoring the positive dynamics of average daily gain as early as on the 5th day after the start of treatment. Haematological studies indicate stimulation of the immune system in animals that was manifested by a 14% increase in the count of lymphocytes and a 37% increase in the proteins of the γ-globulin fraction compared to healthy animals. These indices suggest an increase in the activity of the humoral part of the immune system in animals after treatment with the preparation. Another positive therapeutic effect of the preparation was observed in the protective effect on the liver of diseased animals. Despite the present effect of both preparations, the decrease in the activity of markers characterising hepatocyte destruction was higher with the Kazakh preparation. Thus, the created preparation is characterised by a higher therapeutic effect in comparison with the imported preparation and allow recommending it for treatment and prevention of diarrhoea in young animals

Ribosomal protein L17 functions as an antimicrobial protein in amphioxus

Antimicrobial peptides (AMPs), characterized by their cationic nature and amphiphilic properties, play a pivotal role in inhibiting the biological activity of microbes. Currently, only a fraction of the antimicrobial potential within the ribosomal protein family has been explored, despite its extensive membership and resemblance to AMPs. Herein we demonstrated that amphioxus RPL17 (BjRPL17) exhibited not only upregulated expression upon bacterial stimulation but also possessed bactericidal capabilities against both Gram-negative and -positive bacteria through combined action mechanisms including interaction with cell surface molecules LPS, LTA, and PGN, disruption of cell membrane integrity, promotion of membrane depolarization, and induction of intracellular ROS production. Furthermore, a peptide derived from residues 127–141 of BjRPL17 (termed BjRPL17-1) showed antibacterial activity against Staphylococcus aureus and its methicillin-resistant strain via the same mechanism observed for the full-length protein. Additionally, the rpl17 gene was highly conserved in Metazoa, hinting it may play a universal role in the antibacterial defense system in different animals. Importantly, neither BjRPL17 nor peptide BjRPL17-1 exhibited toxicity towards mammalian cells thereby offering prospects for designing novel AMP agents based on these findings. Collectively, our results establish RPL17 as a novel member of AMPs with remarkable evolutionary conservation.

Effect of the organoselenium compound 974zh on the TLR2 and TLR4 gene expression in blood and spleen cells of experimental animals when co-administered with Yersinia pestis EV

One of the important directions for increasing the immunogenic properties of vaccine strains against highly dangerous infections is the search for adjuvants that not only stimulate the immunological effectiveness of vaccination, but can also provide a metabolic correction of the vaccination process. Organoselenium compounds have immunotropic properties and an antioxidant effect, and therefore, the study of the effect of the organoselenium compound 2,6-dipyridinium-9-selenabicyclo[3.3.1]nonane dibromide (974zh) on the activity of the TLR2 and TLR4 gene expression by macroorganism cells of experimental animals immunized with Yersinia pestis EV NIIEG vaccine strain, is a current area of research.The aim of the work. To assess the TLR2 and TLR4 gene expression by cells of the immune phagocyte system of experimental animals immunized with the Y. pestis EV vaccine strain against the background of immunomodulation with the organoselenium compound 974zh.Materials and methods. The study was carried out on 125 certified outbred white mice. Biological material (blood, spleen) was disinfected, and the spleen was homogenized. RNA isolation and reverse transcription were performed using commercial reagent kits. The expression level of the TLR2 and TLR4 genes was determined using real-time polymerase chain reaction with specific primers.Results. When assessing innate immunity using the example of blood and spleen cells of animal models, features of the TLR2 and TLR4 gene expression were revealed in response to the introduction of the Y. pestis EV vaccine strain against the background of immunomodulation with the 974zh. It was found that 974zh induces a statistically significant increase in TLR2 gene expression when co-administered with Y. pestis EV at a dose of both 104 CFU and 103 CFU.Conclusion. Y. pestis EV against the background of immunomodulation with 974zh, stimulates the expression of the TLR2 and TLR4 genes, which may indicate an increase in the immunogenic properties of the Y. pestis EV vaccine strain under the influence of this preparation.

Citrinin Provoke DNA Damage and Cell-Cycle Arrest Related to Chk2 and FANCD2 Checkpoint Proteins in Hepatocellular and Adenocarcinoma Cell Lines.

Citrinin (CIT), a polyketide mycotoxin produced by Penicillium, Aspergillus, and Monascus species, is a contaminant that has been found in various food commodities and was also detected in house dust. Several studies showed that CIT can impair the kidney, liver, heart, immune, and reproductive systems in animals by mechanisms so far not completely elucidated. In this study, we investigated the CIT mode of action on two human tumor cell lines, HepG2 (hepatocellular carcinoma) and A549 (lung adenocarcinoma). Cytotoxic concentrations were determined using an MTT proliferation assay. The genotoxic effect of sub-IC50 concentrations was investigated using the alkaline comet assay and the impact on the cell cycle using flow cytometry. Additionally, the CIT effect on the total amount and phosphorylation of two cell-cycle-checkpoint proteins, the serine/threonine kinase Chk2 and Fanconi anemia (FA) group D2 (FANCD2), was determined by the cell-based ELISA. The data were analyzed using GraphPad Prism statistical software. The CIT IC50 for HepG2 was 107.3 µM, and for A549, it was >250 µM. The results showed that sensitivity to CIT is cell-type dependent and that CIT in sub-IC50 and near IC50 induces significant DNA damage and cell-cycle arrest in the G2/M phase, which is related to the increase in total and phosphorylated Chk2 and FANCD2 checkpoint proteins in HepG2 and A549 cells.

The Integrative Life history of Maternal Effects.

Context-dependent allocation of resources drives trade-offs among fitness-related traits and other phenotypes to which those traits are linked. In addition, the amount and type of acquired resources can also affect the phenotypes of other organisms through indirect genetic effects, as exemplified by the maternal provisioning of offspring. Despite a large literature on maternal effects, we lack a comprehensive understanding of the extent to which mothers might affect the phenotypes of their offspring, as well as the various mechanisms by which they do so, particularly with regard to many functional traits that are key determinants of survival and reproduction. Our goals in this paper are to review the various approaches to measuring and understanding maternal effects, and to highlight some promising avenues for integration of maternal effects with some other key areas of evolutionary ecology. We focus especially on nutritional geometry; maternal age; and traits proximate to fitness such as whole-organism performance. Finally, we discuss the logistic and practical limits of quantifying these effects in many animal systems, and emphasize the value of integrative approaches in understanding the mechanisms underlying maternal influence on offspring phenotypes.

Through an animal's eye: the implications of diverse sensory systems in scientific experimentation.

'Accounting for the sensory abilities of animals is critical in experimental design.' No researcher would disagree with this statement, yet it is often the case that we inadvertently fall for anthropocentric biases and use ourselves as the reference point. This paper discusses the risks of adopting an anthropocentric view when working with non-human animals, and the unintended consequences this has on our experimental designs and results. To this aim, we provide general examples of anthropocentric bias from different fields of animal research, with a particular focus on animal cognition and behaviour, and lay out the potential consequences of adopting a human-based perspective. Knowledge of the sensory abilities, both in terms of similarities to humans and peculiarities of the investigated species, is crucial to ensure solid conclusions. A more careful consideration of the diverse sensory systems of animals would improve many scientific fields and enhance animal welfare in the laboratory.

Emamectin benzoate exposure induced carp kidney injury by triggering mitochondrial oxidative stress to accelerate ferroptosis and autophagy

Emamectin benzoate (EMB), commonly used as an insecticide in fishery production, inevitably leaves residual chemicals in aquatic environments. High-level EMB exposure can cause severe damage to multiple systems of marine animals, potentially through mechanisms involving severe mitochondrial damage and oxidative stress. However, it is not clear yet how EMB exposure at a certain level can cause damage to fish kidney tissue. In this study, we exposed carps to an aquatic environment containing 2.4 μg/L of EMB and cultured carp kidney cells in vitro, established a cell model exposed to EMB. Our findings revealed that EMB exposure resulted in severe kidney tissue damage in carp and compromised the viability of grass carp kidney cells (CIK cells). By RNA-seq analysis, EMB exposure led to significant differences in mitochondrial homeostasis, response to ROS, ferroptosis, and autophagy signals in carp kidney tissue. Mechanistically, EMB exposure induced mitochondrial oxidative stress by promoting the generation of mitochondrial superoxide and reducing the activity of antioxidant enzymes. Additionally, EMB exposure triggered loss of mitochondrial membrane potential, an imbalance in mitochondrial fusion/division homeostasis, and dysfunction in oxidative phosphorylation, ultimately impairing ATP synthesis. Notably, EMB exposure also accelerated excessive autophagy and ferroptosis of cells by contributing to the formation of lipid peroxides and autophagosomes, and the deposition of Fe2+. However, N-acetyl-L-cysteine (NAC) treatment alleviated the damage and death of CIK cells by inhibiting oxidative stress. Overall, our study demonstrated that EMB exposure induced mitochondrial oxidative stress, impaired mitochondrial homeostasis, and function, promoted autophagy and ferroptosis of kidney cells, and ultimately led to kidney tissue damage in carp. Our research enhanced the toxicological understanding on EMB exposure and provides a model reference for comparative medicine.

Investigation through Animal-Computer Interaction: A Proof-of-Concept Study for the Behavioural Experimentation of Colour Vision in Zoo-Housed Primates.

Zoos are an important repository of animals, which have a wide range of visual systems, providing excellent opportunities to investigate many comparative questions in sensory ecology. However, behavioural testing must be carried out in an animal welfare-friendly manner, which is practical for zoo staff. Here, we present a proof-of-concept study to facilitate behavioural research on the sensory ecology of captive primates. A system consisting of a tablet computer and an automated feeder connected wirelessly was developed and presented to captive primate species to evaluate interactions with and without previous training. A colour stimulus, analogous to the Ishihara test, was used to check the level of interaction with the device, supporting future studies on sensory ecology with zoo animals. Animals were able to use the system successfully and displayed signs of learning to discriminate between the visual stimuli presented. We identified no risk for small primates in their interactions with the experimental setup without the presence of keepers. The use of electronic devices should be approached with caution to prevent accidents, as a standard practice for environmental enrichment for larger animals (e.g., spider monkeys). In the long term, the system developed here will allow us to address complex comparative questions about the functions of different visual systems in captive animals (i.e., dichromatic, trichromatic, etc.).

Asymptomatic herpes simplex virus brain infection elicits cellular senescence phenotypes in the central nervous system of mice suffering multiple sclerosis-like disease

Experimental autoimmune encephalomyelitis (EAE) is a demyelinating disease affecting the central nervous system (CNS) in animals that parallels several clinical and molecular traits of multiple sclerosis in humans. Herpes simplex virus type 1 (HSV-1) infection mainly causes cold sores and eye diseases, yet eventually, it can also reach the CNS, leading to acute encephalitis. Notably, a significant proportion of healthy individuals are likely to have asymptomatic HSV-1 brain infection with chronic brain inflammation due to persistent latent infection in neurons. Because cellular senescence is suggested as a potential factor contributing to the development of various neurodegenerative disorders, including multiple sclerosis, and viral infections may induce a premature senescence state in the CNS, potentially increasing susceptibility to such disorders, here we examine the presence of senescence-related markers in the brains and spinal cords of mice with asymptomatic HSV-1 brain infection, EAE, and both conditions. Across all scenarios, we find a significant increases of senescence biomarkers in the CNS with some differences depending on the analyzed group. Notably, some senescence biomarkers are exclusively observed in mice with the combined conditions. These results indicate that asymptomatic HSV-1 brain infection and EAE associate with a significant expression of senescence biomarkers in the CNS.

Chlamydomonas

Tulin et al. introduce Chlamydomonas, a unicellular green alga commonly used as a microbial reference system for plants and animals.

Current status and challenges in disease surveillance and epidemiological investigation systems for companion animals in South Korea

Current status and challenges in disease surveillance and epidemiological investigation systems for companion animals in South Korea

State regulation of insurance in agriculture: global experience, analysis of the current situation, prospects

The work focuses on the role of insurance to support sustainable agricultural production in economically developed countries, the importance of public-private partnerships in accordance with the potential of agro-industrial complex. Purpose – mechanism of state regulation of insurance of agricultural risks is considered using the example of the world experience, taking into account the situation in Kazakhstan. Methods – comparison and scientific generalization, review of literature sources on the topic. The study is divided into two stages: analysis of statistical sources on agricultural insurance at the international level from 2020 to 2023; study of domestic practice. Results – advantages and disadvantages of various approaches to insurance of agricultural activities with state participation have been identified. Recommendations for improving the insurance system of agricultural crops and animals in the republic are substantiated, including precise definition of responsibilities and conditions of participation for insurance firms, increasing the level of subsidizing premiums, ensuring the availability of information for farmers and creating a legislative framework. The work also notes the need to adapt international experience to Kazakhstan’s national characteristics. Conclusions - state support for compensation of damage caused by natural disasters, the implementation of the proposed measures will significantly increase the efficiency of agricultural insurance in the country, ensure protection of the interests of farms, stable development of agricultural sector and minimize risks for agricultural producers. The legal mechanism of insurance in agro-industrial complex requires improvement. As a basis for regulation, it is necessary to establish constructive dialogue between stakeholders. To assess the condition of agricultural crops, the use of space monitoring data is of great importance. Agricultural insurance is a potential growth segment and factor in food security in Kazakhstan.

DNA Virome in Cardiac Tissue from Green Sea Turtles (Chelonia mydas) with Myocarditis.

As part of a sea turtle health monitoring program on the central east coast of Queensland, Australia, stranded and sick green sea turtles (Chelonia mydas) were subjected to necropsy and histopathology. A subset of these turtles had myocarditis of varying severity, which could not be attributed to parasitism by spirorchid flukes or bacterial infections. We, therefore, undertook an investigation to determine whether virus infections might be part of the pathogenesis. Deep sequencing revealed abundant DNA virus contigs in the heart tissue, of which CRESS and circoviruses appeared to be the most consistently present. Further analysis revealed the homology of some of the circoviruses to the beak and feather disease virus. While a causative link to myocarditis could not be established, the presence of these viruses may play a contributing role by affecting the immune system and overall health of animals exposed to pollutants, higher water temperatures, and decreasing nutrition.

Minocycline Inhibits Tick-Borne Encephalitis Virus and Protects Infected Cells via Multiple Pathways.

Tick-borne Encephalitis (TBE) is a zoonotic disease caused by the Tick-borne Encephalitis virus (TBEV), which affects the central nervous system of both humans and animals. Currently, there is no specific therapy for patients with TBE, with symptomatic treatment being the primary approach. In this study, the effects of minocycline (MIN), which is a kind of tetracycline antibiotic, on TBEV propagation and cellular protection in TBEV-infected cell lines were evaluated. Indirect immunofluorescence, virus titers, and RT-qPCR results showed that 48 h post-treatment with MIN, TBEV replication was significantly inhibited in a dose-dependent manner. In addition, the inhibitory effect of MIN on different TBEV multiplicities of infection (MOIs) in Vero cells was studied. Furthermore, the transcriptomic analysis and RT-qPCR results indicate that after incubation with MIN, the levels of TBEV and CALML4 were decreased, whereas the levels of calcium channel receptors, such as RYR2 and SNAP25, were significantly increased. MIN also regulated MAPK-ERK-related factors, including FGF2, PDGFRA, PLCB2, and p-ERK, and inhibited inflammatory responses. These data indicate that administering MIN to TBEV-infected cells can reduce the TBEV level, regulate calcium signaling pathway-associated proteins, and inhibit the MAPK-ERK signaling pathway and inflammatory responses. This research offers innovative strategies for the advancement of anti-TBEV therapy.

Comparative connectomics of the descending and ascending neurons of the Drosophila nervous system: stereotypy and sexual dimorphism.

In most complex nervous systems there is a clear anatomical separation between the nerve cord, which contains most of the final motor outputs necessary for behaviour, and the brain. In insects, the neck connective is both a physical and information bottleneck connecting the brain and the ventral nerve cord (VNC, spinal cord analogue) and comprises diverse populations of descending (DN), ascending (AN) and sensory ascending neurons, which are crucial for sensorimotor signalling and control. Integrating three separate EM datasets, we now provide a complete connectomic description of the ascending and descending neurons of the female nervous system of Drosophila and compare them with neurons of the male nerve cord. Proofread neuronal reconstructions have been matched across hemispheres, datasets and sexes. Crucially, we have also matched 51% of DN cell types to light level data defining specific driver lines as well as classifying all ascending populations. We use these results to reveal the general architecture, tracts, neuropil innervation and connectivity of neck connective neurons. We observe connected chains of descending and ascending neurons spanning the neck, which may subserve motor sequences. We provide a complete description of sexually dimorphic DN and AN populations, with detailed analysis of circuits implicated in sex-related behaviours, including female ovipositor extrusion (DNp13), male courtship (DNa12/aSP22) and song production (AN hemilineage 08B). Our work represents the first EM-level circuit analyses spanning the entire central nervous system of an adult animal.

Peculiarities of the blood system of animals under the conditions of consumption of drinking water containing chlorates

In modern conditions, providing the population with high-quality drinking water is becoming an increasingly urgent hygienic, scientific, technical, and social problem. Chlorine dioxide can be an alternative reagent for disinfecting drinking water. However, as with chlorine, dangerous by-products, particularly chlorites and chlorates are formed. The processes of development of morphofunctional changes in homeostasis remain insufficiently studied in this aspect. The objective is to determine the nature and peculiarities of changes in hematological indicators in the blood of animals due to the chronic impact of various concentrations of chlorates in drinking water to establish criteria for the hygienic assessment of their adverse effects on the body. The test animals were given drinking water with different concentrations of chlorates daily for 6 months: group 1 – control, group 2 – chlorates at a dose of 0.2 mg/dm³, group 3 – chlorates at a dose of 0.7 mg/dm³, and group 4 – chlorates at a dose of 1.2 mg/dm³. Evaluating the results of experimental studies, it should be noted that the most pronounced changes are observed in groups of animals that received chlorates in concentrations of 0.7 and 1.2 mg/dm³: a gradual decrease in the absolute number of leukocytes, the absolute and relative number of lymphocytes, relative to the indicators of the control group can be observed. In groups of animals that received chlorates in concentrations of 0.7 and 1.2 mg/dm³, a decrease in the absolute number of erythrocytes was observed during the 90 days of the experiment. It was established that the severity of the effects depended on the dose of chlorates and the time of exposure. Thus, the nature and features of changes in hematological indicators in the body of experimental animals during a chronic sanitary-toxicological experiment under the influence of chlorates in drinking water may indicate that chlorates affect the indicators of the blood system, being the indirect signs of disorders of other functional systems in the body, namely – slowing of redox reactions, hypoxic manifestations, reduction and weakening of the immune response and reactivity, etc.

Determination of the optimal concentration of ozone–containing mixture for safe use in clinical conditions

Objective. To study the effect of ozone during hyperbaric oxygenation with an ozone–containing mixture on the respiratory system and stress response organs of experimental animals, to select the optimal parameters of ozone generation and supply by the POS–1 apparatus to perform this procedure in a confined space without harming the health of the patient and medical staff. Materials and methods. Experimental studies were performed on 20 white laboratory rats divided into two groups. In group 1, 10 animals breathed the ozone–containing mixture for 7 days, each session lasted 15 minutes. In the 2nd group, 10 animals breathed the ozone–containing vapour–water mixture for 7 days, each session also lasted 15 minutes. All animals were kept in a semi–hermetic box with a volume of 0.042 m3 with an ozone supply of 500 mg/h, which exceeds the maximum permissible concentration (0.16 mg/m3 ) by a factor of 2. The pressure of the gas mixture in the box with the animals was equal to atmospheric pressure.The animals were withdrawn from the experiment on the 3rd, 7th and 14th day by injecting an excessive dose of 5.0% sodium thiopental solution. The trachea, lungs, liver, kidneys, spleen, heart, stomach, pancreas, and adrenal glands were autopsied and taken for histological examination. The ozone concentration in the room during the experiment was measured using a SAMI 100S ozone concentration meter. Results. Exposure to the ozone–containing mixture did not cause pathological changes in the organs and systems of the experimental animals, as well as a significant stress response, which proves the safety of the technique in confined spaces. The method of treatment of infectious and purulent complications proposed in previous studies does not harm the health of the patient and medical staff and can be recommended for use in clinical practice without additional measures for the utilisation of ozone produced by the POS–1 apparatus. Conclusions. The optimal parameters of ozone production and supply in the ozone–containing mixture for hyperbaric oxygenation in a confined space without harming the health of the patient and medical staff have been proposed. The ozone–containing steam–water mixture can be recommended for use in clinical practice in conditions of massive tissue infection as a factor of external physical influence with pronounced bactericidal and bacteriostatic properties. The concentration of residual ozone in the room after the therapeutic procedure does not exceed the permissible level and does not harm the health of the patient and medical staff.

Herbal Hair Serum – A Promising Nourishing Agent for the Hair Growth

With their promise of nourishing properties, improved scalp health, and stimulation of hair growth, herbal hair serums have become increasingly popular as natural substitutes for traditional hair care products. An overview of herbal hair serums is given in this abstract, with particular attention to their composition, effectiveness, safety, and possible advantages for hair health to the assertion of reduced hair loss and improved hair growth. Alopecia is a common issue among metropolitan dwellers who are subjected to stress, environmental issues, and other issues. Thus, we deduce from this review paper that alopecia may be effectively treated with numerous herbal remedies without any adverse effects. There are several allopathic medications kinds for treating hair loss, however they all have a lot of adverse effects. The foundation of any medical research is its herb collection. About 80% of residents suggested using herbal medications since they had fewer negative effects and more positive effects than the animal system. Our article shows that herbal hair serum contains a number of vital ingredients that are necessary to maintain the correct operation and self-defense. It also offers excellent protection from extreme heat. Many factors have contributed to the severe hair loss issues that the younger generations are now experiencing. Generally speaking, hair loss is not transient. Known to be the most significant organ in the mammalian system, the hair follicle has a variety of roles in self-defense, extreme temperature protection, and attractiveness and gender differentiation. The younger generations are facing severe hair loss issues as a result of several lifestyle changes, including worry, weariness, and overindulgence in junk food and hair colouring and styling products.t alopecia is the outcome. To encourage the production of new hair and prevention of hair loss, hair roots must be activated which is why many individuals experiencing hair loss are seeking different treatments. The anti-dandruff and hair care properties of Citrus sinensis are employed