Dermatophytosis In Animals Research Articles

Studies on the in vitro mechanism and in vivo therapeutic effect of the antimicrobial peptide ACP5 against Trichophyton mentagrophytes

Trichophyton mentagrophytes is a zoophilic dermatophyte that can cause dermatophytosis in humans and animals. Antimicrobial peptides (AMPs) are considered as a promising agent to overcome the drug-resistance of T. mentagrophytes. Our findings suggest that cationic antimicrobial peptide (ACP5) not only possesses stronger activity against T. mentagrophytes than fluconazole, but also shows lower toxicity to L929 mouse fibroblast cells than terbinafine. Notably, its resistance development rate after resistance induction was lower than terbinafine. The present study aimed to evaluate the fungicidal mechanism of ACP5 in vitro and its potential to treat dermatophyte infections in vivo. ACP5 at 1 ×MIC completely inhibited T. mentagrophytes spore germination in vitro. ACP5 severely disrupts the mycelial morphology, leading to mycelial rupture. Mechanistically, ACP5 induces excessive ROS production, damaging the integrity of the cell membrane and decreasing the mitochondrial membrane potential, causing irreversible damage in T. mentagrophytes. Furthermore, 1% ACP5 showed similar efficacy to the commercially available drug 1% terbinafine in a guinea pig dermatophytosis model, and the complete eradication of T. mentagrophytes from the skin by ACP5 was verified by tissue section observation. These results indicate that ACP5 is a promising candidate for the development of new agent to combat dermatophyte resistance.

РАСПРОСТРАНЕНИЕ И СИМПТОМАТИКА ДЕРМАТОМИКОЗОВ ЖИВОТНЫХ В КРАСНОДАРСКОМ КРАЕ

Epizootological examinations of livestock enterprises in the Krasnodar Territory and veterinary clinics in Krasnodar have shown that skin diseases are often registered in agricultural and domestic animals. The incidence of skin diseases in domestic animals is significantly higher than in agricultural animals. In farm animals, ringworm is recorded in the range of 2-5%, in domestic carnivorous animals, dermatophytosis is found in the range of 50-60%, most often in unvaccinated animals. Dermatomycosis in farm animals revealed a deep form of 48.0%, an atypical 37.0% and a superficial form of the disease of 16.0%. The economic damage is significant, resulting from a decrease in productivity, the cost of diagnostic, therapeutic drugs and necessary vaccines. Ringworm is manifested in farm animals by inflammation of the skin on the trunk, head, neck and distal part of the limbs. Dermatophytosis in carnivorous animals is characterized by inflammatory processes on the head, eyes, ears, neck, trunk and upper extremities. The degree of damage depends on the type of pathogen, dose, and place of introduction. Microsporia is more common from 5 to 7% than trichophytia 3.5 to 4.5%, with age the incidence in carnivores decreases, after 8 years dermatomycosis is less common, trichophytia 2.1%, and microsporia 1.9%. Dermatomycosis in farm animals revealed a deep form (48.0%), atypical (37.0%) and superficial form of the disease (16.0%). Dermatophytosis mainly affects young carnivores, the peak incidence is manifested at the age of 8 to 18 months. The maximum number of infected animals with dermatophytes is observed in the month of October at 30.5%.

The use of propolis in ointment ingredient for the treatment of dermatophytosis infection

Dermatophytosis in animals is very important since it is zoonotic. Propolis is a promising natural compound against pathogenic fungi since there is an increasing need to discover less expensive, more natural, and more accessible treatments. The purpose of this study was to evaluate propolis as an anti-dermatophyte and topical drug preparation against Trichophyton mentagrophytes. Propolis was tested for its antifungal properties at various concentrations using the diffusion method. The ointment was applied to the rabbit with dermatophytosis after being prepared by combining the ointment base with 10% propolis. The result demonstrated that propolis was able to inhibit the growth of Trichophyton mentagrophytes which was indicated by a clear zone around the disc. Propolis ointment helped lessen inflammation more than Miconazole 2% at severe lesions, according to in vivo trials on rabbits, while the two ointment’s healing times were nearly identical. According to the findings, propolis is a promising ointment ingredient for the treatment of dermatophytosis.

Metabolic and morphostructural plasticity of environmental and animal strains of Nannizzia gypsea.

Nannizzia gypsea is a geophylic agent of human and animal dermatophytosis. This study compares the metabolic and morphostructural plasticity of N. gypsea strains isolated from moss, sand, and a dog. The in vitro metabolic plasticity included the detection of extracellular enzymes, thermotolerance, resistance to oxidative stress, and assessment of fungal growth. Structural plasticity studies included cell surface hydrophobicity, electronegativity, and size of macroconidia. Virulence was assessed on Tenebrio mollitor model. The strains showed low thermotolerance, susceptibility to oxidative stress, and were producers of keratinase, lipase and catalase. N. gypsea strains were unable to produce hemolysin, esterase, and phospholipase although they were able to grow with different carbon sources. The electronegative properties of the surface did not vary between the strains under study. The knowledge about N. gypsea metabolic and morphostructural plasticity could be crucial for the development of therapeutic strategies and control of dermatophytosis.

Comparative assessment of regulated methods and PCR in the diagnosis of trichophytosis in veterinary mycology.

There is an increase in the incidence of human and animal infectious skin diseases of fungal etiology in the world. The main source of infecting the population has become agricultural and stray animals. The objective of this study was to examine the morphophysiological and microbiological characteristics of pathogenic fungi belonging to the species Trichophyton verrucosum. This species is known to cause diseases in both humans and livestock in Kazakhstan. In addition, the study aimed to assess the feasibility of using the polymerase chain reaction (PCR) method for detecting T. verrucosum. This assessment was conducted in comparison to the outcomes of conventional laboratory diagnostic tests commonly employed for trichophytosis. The research focused on analyzing 141 samples of pathological material obtained from calves in Almaty, Turkestan, and Kyzylorda regions. These calves exhibited clinical symptoms of skin disease. The study aimed to identify the causative agent using various techniques, including microscopic examination, microbiological methods involving the isolation of pure cultures, and PCR. The detection of the causative agent of dermatophytosis using conventional methods was relatively low, 86% for the microscopic method, and 79% for the microbiological method with the isolation of the culture of the pathogen. Extraction and detection of the genetic material of the causative agent of the disease for PCR was carried out according to the method developed by the authors. The effectiveness of the PCR method was 97.9%, which is significantly higher (p < 0.05) compared with the diagnostic effectiveness of conventional methods. The PCR method using specific primers identified the causative agent in 98% of cases, which significantly (p < 0.05) exceeded the results obtained using conventional diagnostic methods. Accordingly, the PCR method had better sensitivity and specificity indicators. The conducted study recommends the method of PCR diagnosis of dermatophytosis for fast and reliable confirmation of the diagnosis of dermatophytosis in humans and animals in Kazakhstan.

Descriptive epidemiology of dermatophytosis in rodents.

Dermatophytosis is a zoonotic disease caused by a group of keratinophilic fungi called dermatophytes. Since the epidemiology of diseases revolves over time, this research studies the incidence of dermatophytosis among rodents referred to mycology laboratory during 2019-2021. A total of 163 rodents including rabbits, guinea pigs, and hamsters suspecting having dermatophytosis were sampled by scraping lesions. Direct microscopic examination, culture, and polymerase chain reaction were done for diagnosis of dermatophytosis and identification of the etiologic agent. The results of this study showed that 37.4% of rodents were involved with dermatophytosis, among which 41.13% of rabbits, 25% of guinea pigs, and 26.3% of hamsters were included. Microsporum canis (52.7%) was the most isolated agent. Incidence of dermatophytosis was higher in female in rabbits while in hamsters and guinea pigs male were mostly infected. Rodents less than 6 months were more susceptible for dermatophytosis except for hamsters in which 6-12 months animals had a higher prevalence. In conclusion, it is significant to update our knowledge about the epidemiology of dermatophytosis in rodents and other animals every few years to define valid preventive strategies. Moreover, since dermatophytes are contagious and zoonotic, it is also a priority to apply preventing methods for dermatophytosis and treat infected rodents with appropriate antifungal agents to decrease the risk of infection.

Effects of Coleus amboinicus L. Essential Oil and Ethanolic Extracts on Planktonic Cells and Biofilm Formation of Microsporum canis Isolated from Feline Dermatophytosis.

Microsporum canis is an important zoonotic fungus that causes dermatophytosis in domestic animals and their owners. Domestic cats are the primary reservoir for M. canis. Antifungal drugs frequently produce adverse effects on the host animal, increasing the demand for novel alternative treatments derived from nature. We evaluated the antifungal activity of Coleus amboinicus essential oil (CEO) and ethanolic extracts (CEE) against M. canis in planktonic and biofilm growth. Twelve clinical isolates of M. canis were identified in feline dermatophyte samples. Using GC-MS, 18 compounds were identified in CEO, with carvacrol being the major constituent. HPLC analysis of CEE revealed that it contained rosmarinic acid, apigenin, and caffeic acid. The planktonic growth of all M. canis isolates was inhibited by C. amboinicus extracts. The minimum inhibitory concentration at which ≥50% of the isolates were inhibited (MIC50) was 128 µg/mL (32-256 µg/mL) for both CEO and CEE. The MIC90 values of CEO and CEE were 128 and 256 µg/mL, respectively. CEO at MIC (128 µg/mL) and 2× MIC (256 µg/mL) significantly inhibited the biofilm formation of weak, moderate, and strong biofilm-producing M. canis. CEE at 2× MIC (256 µg/mL) significantly inhibited the biofilm formation of all isolates. Overall, C. amboinicus extracts inhibited planktonic growth and exhibited a significant antibiofilm effect against M. canis. Thus, C. amboinicus is a potential source of natural antifungal compounds.

Study of the effectiveness of a new antifungal agent for the treatment of animal dermatophytosis

Relevance. Small domestic animals often suffer from dermatophytosis chronically, while remaining active carriers. In clinical medicine, there is a certain arsenal of drugs with antifungal activity, both ofsystemic and local action, the use of which often causes side effects. In this regard, the search, development and implementation in public health and veterinary practice of highly active and low-toxic drugs that can specifically affect pathogenic fungi without adverse effects on the body is relevant. On the basis of screening microbiological studies, a thiadiazole derivative was selected, which in preliminary experiments showed antifungal activity.Methods. The study was conducted on the basis of the St. Petersburg State Chemical Pharmaceutical University for 21 days. 19 cats in the analyzes of which a fungus Microsporum canis was detectedwere used in the experiment. An ointment based on a thiadiazole derivative was rubbed into the affected area of the skin once a day in an amount of 0.01–0.05. The therapeutic efficacy of ointments was evaluated for 21 days according to the following parameters: specific glow of lesions in the rays of a Wood's lamp in the presence of a pathogen; results ofseeding and microscopic examination of the material; coat recovery time; local irritant action.Results. When animals were treated with 3% thiadiazole ointment, on the 10–12th day the inflammation regressed, and abundant small-lamellar peeling was observed on the skin in the foci. By 12–15th day the surface of the foci was cleared, and the skin acquired a pale pink color. The restoration of the coat was observed at the end of the application of the ointment — on 21st day. The specific glow of regrown hairs in the rays of the Wood's lamp ceased. The results of seeding and microscopy of regrown hairs became negative. Negative reactions when applying the ointment to the skin of animals were not observed. The results of the study indicate high efficacy and good tolerability of 3% thiadiazole ointment.

S5.4c Isolation of keratinophilic fungi of the genus Microsporum from the soils of Moscow, Russia

5.4 Free oral paper session, September 22, 2022, 3:00 PM - 4:30 PMIt is reported about the discovery of zoophilic and geophilic dermatophytes in the soils of city parks and adjacent territories of Moscow, Russia. Microscopic fungi of the Microsporum genus were found in 27/50 samples (54%): Microsporum gypseum (42%) and Microsporum canis (12%), their number was higher in the soils taken from the adjacent areas where domestic animals are walked. This increases the risk of infection of animals upon contact with soil.ObjectivesThe research was conducted to determine the frequency of occurrence of potential pathogens of animal dermatophytosis in those areas of the soil cover where pets are walking. Soil samples were taken from several areas: the adjacent territories along the streets of Vishnevsky and Kostyakov, designated as areas 1 and 2, the park ‘Dubki’, Lianozovsky, Timiryazevsky, designated as 3, 4, and 5, respectively.Areas 1 and 2 are narrow patches of soil adjacent to residential buildings and sidewalks where pets are most often walked. Areas 3 and 4 are areas of soil adjacent to park paths, where animal walking is less common. Area 5 is a relatively little-visited place in the forest park, where animals are rarely walked.MethodsBefore sampling, vegetation cover and rubbish were removed from the soil area, then ∼ 100 g of soil was taken from the surface using aseptic rules in sterile plastic containers. A total of 10 soil samples were taken from each plot. The soil was dried at room temperature, sieved to remove rubbish, and plant parts, and placed in sterile Petri dishes.Isolation of fungi was carried out by the ‘bait’ method: samples of sterile pet hair about 1 cm long were placed on the surface of the soil, the soil was moistened with sterile distilled water with the addition of a selective additive for the isolation of dermatophytes (Dermasel, Oxoid). Petri dishes were periodically inspected for fungal growth on the surface of the wool and moistened if necessary.When the growth of microscopic fungi was detected on the surface of the wool, they were sterilely transferred to Petri dishes with wort agar medium (M129, HiMedia) to obtain a pure culture and identification. Only microscopic fungi of the genus Microsporum were taken into account.ResultsAs a result of the research, a total of 27 isolates of microscopic fungi, 21 of Microsporum gypseum and 6 of Microsporum canis were isolated from the soils of Moscow: 7 isolates at the area 1 (6 of M. gypseum, 1 of M. canis), 8 isolates at the area 2 (5 of M. gypseum, 3 of M. canis), 5 isolates at areas 3, and 5 (4 of M. gypseum, 1 of M. canis), 2 isolates of M. gypseum at the area 5. The results are illustrated in Figure 1.ConclusionSoils located in adjacent areas are more polluted by the hair of pets that walk there. Therefore, these soils also contain keratinophilic microscopic fungi, which includes the pathogens of animal dermatophytosis. In such places, animals are susceptible to infection with fungal infections directly from the soil.

The Use of Pythium oligandrum in the Biological Control of Roundworm Infection in Dogs and Cats.

Pythium oligandrum is an oomycete used in veterinary medicine to treat dermatophytosis in animals. The ovicidal potential against various types of parasite eggs has not been investigated. Ascarids are very common in young animals and the resistance of eggs in the external environment is very high. A commercial product containing P. oligandrum was used in the present study. Its ovicidal action against Toxocara spp. eggs was investigated. Thus, two categories of media (soil and sandstone) were used, on which the ascarid eggs were dispersed. The commercial product was prepared as a solution and was applied according to the manufacturer. The treatment scheme indicated in case of animals with dermatophytosis was used. Thus, the external natural conditions were recreated in the laboratory, in order to establish whether there is any applicability for this oomycete in the biological control of some parasitic diseases. The results indicated an ovicidal action of Pythium oligandrum, supporting the prospects of its use in the decontamination methods of various surfaces or environments where ascarid eggs from carnivores are found.

An ancient haplotype containing antimicrobial peptide gene variants is associated with severe fungal skin disease in Persian cats.

Dermatophytosis, also known as ringworm, is a contagious fungal skin disease affecting humans and animals worldwide. Persian cats exhibit severe forms of the disease more commonly than other breeds of cat, including other long-haired breeds. Certain types of severe dermatophytosis in humans are reportedly caused by monogenic inborn errors of immunity. The goal of this study was to identify genetic variants in Persian cats contributing to the phenotype of severe dermatophytosis. Whole-genome sequencing of case and control Persian cats followed by a genome-wide association study identified a highly divergent, disease-associated haplotype on chromosome F1 containing the S100 family of genes. S100 calcium binding protein A9 (S100A9), which encodes a subunit of the antimicrobial heterodimer known as calprotectin, contained 13 nonsynonymous variants between cases and controls. Evolutionary analysis of S100A9 haplotypes comparing cases, controls, and wild felids suggested the divergent disease-associated haplotype was likely introgressed into the domestic cat lineage and maintained via balancing selection. We demonstrated marked upregulation of calprotectin expression in the feline epidermis during dermatophytosis, suggesting involvement in disease pathogenesis. Given this divergent allele has been maintained in domestic cat and wildcat populations, this haplotype may have beneficial effects against other pathogens. The pathogen specificity of this altered protein should be investigated before attempting to reduce the allele frequency in the Persian cat breed. Further work is needed to clarify if severe Persian dermatophytosis is a monogenic disease or if hidden disease-susceptibility loci remain to be discovered. Consideration should be given to engineering antimicrobial peptides such as calprotectin for topical treatment of dermatophytosis in humans and animals.

RNA-Seq Analysis of the Effect of Zinc Deficiency on Microsporum canis, ZafA Gene Is Important for Growth and Pathogenicity.

Microsporum canis, a common pathogenic skin fungus, can cause dermatophytosis in humans and animals. Zinc is an important trace element and plays an important role in the growth and metabolism of fungi. Currently, the effects of zinc deficiency on growth, gene expression, and metabolic pathway have not been clarified in M. canis. Therefore, M. canis was cultured under zinc restriction, and RNA-Seq was conducted in this study. The growth of M. canis was severely inhibited, and many genes showed significant upregulation and downregulation in M. canis with zinc deficiency. Zinc deficiency could negatively affect the gene expression and biological metabolic pathway in M. canis. The zinc-responsiveness transcriptional activator (ZafA) gene was significantly upregulated and shared homology with Zap1. Thus, the ZafA gene might be the main transcription factor regulating M. canis zinc homeostasis. The ZafA gene knockout strain, ZafA-hph, was constructed via Agrobacterium tumefaciens-mediated transformation (ATMT) in M. canis for the first time to assess its function. In vitro growth ability, hair biodegradation ability, virulence test, and zinc absorption capacity in ZafA-hph and wild-type M. canis strains were compared. Results showed that the ZafA gene plays an important role in zinc absorption, expression of zinc transporter genes, and growth and pathogenicity in M. canis and can be used as a new drug target. Cutting off the zinc absorption pathway can be used as a way to prevent and control infection in M. canis.

Mycotic infections in animals in India: An update

This review traces the early records of mycotic infections in India, and presents an update of animal mycoses reported from several parts of India. The types of mycoses covered are the dermatophytosis (ringworm) in domestic animals due to well-known species of zoophilic dermatophytes, viz. Trichophyton simii, T. mentagrophytes, T. verrucosum, Microsporum canis and M. nanum, and the geophilic dermatophyte, M. gypseum, Aspergillus spp, Cryptococcus species and other yeast like fungi, histoplasma and blastomyces. The brief clinical and demographic features of infections in different animals are described. A particularly noteworthy finding in literature search is the report of clinical infections in one dog and two cows by an anthropophilic dermatophyte, T. rubrum from Belgachia, Kolkata, West Bengal in 1954. Veterinary scientists are urged to investigate the possible occurrence of infections in animals due to other pathogenic fungi including the dimorphic ones like Histoplasma and Blastomyces.

In vitro virulence evaluation of clinical and environmental isolates of dermatophyte fungi

Dermatophytes are keratinophilic fungi and the causative agent of dermatophytosis in animals and people. In the pathogenesis of this disease, enzymes such as DNase, gelatinase, lipase, keratinase, elastase, and collagenase are highlighted. This work aimed to verify the production of these enzymes by clinical and environmental isolates of dermatophytes. Environmental strains were obtained by the Vanbreuseghem technique (1952), using soil samples from different Brazilian locations. The clinical samples were obtained from animal hair and crust sent to the Veterinary Microbiological Diagnostic Service/UFRRJ. The enzymatic evaluation of the dermatophytes was made by spectrophotometer absorbance readings (collagenase, elastase, and keratinase), degradation halo formation in Petri dishes (DNase and lipase) and tube liquefaction (gelatinase). The clinical isolates were Microsporum canis (11), Nannizzia gypsea (7), N. nana (2), Trichophyton mentagrophytes (4) and Trichophyton sp. (6). The environmental isolates were N. gypsea (25), N. nana (1) and Trichophyton sp. (4). There was no statistically significant difference in keratinase, elastase, lipase and gelatinase production between the clinical and environmental isolates groups. There was a statistically significant difference in collagenase and DNase production. It is concluded that both clinical and soil samples are capable of producing enzymes related to dermatophyte infection.

In vitro antifungal susceptibility patterns of Trichophyton benhamiae complex isolates from diverse origin.

Species from the Trichophyton benhamiae complex are mostly zoophilic dermatophytes which cause inflammatory dermatophytosis in animals and humans worldwide. This study was purposed to (a) to identify 169 reference and clinical dermatophyte strains from the Tbenhamiae complex species by molecular method and adhering to the newest taxonomy in the complex (b) to evaluate the in vitro antifungal susceptibility profile of these strains against eight common and new antifungal agents that may be used for the treatment of dermatophytosis. All isolates, mainly originated from Europe but also from Iran, Japan and USA, were subjected to ITS-rDNA sequencing. The in vitro antifungal susceptibility profiles of eight common and new antifungal drugs against the isolates were determined by CLSI M38-A2 protocol and according to microdilution method. Based on the ITS-rDNA sequencing, Tbenhamiae was the dominant species (n=102), followed by Teuropaeum (n=29), Terinacei (n=23), Tjaponicum (n=10), Trichophyton sp (n=4) and Teriotrephon (n=1). MIC ranges across all isolates were as follows: luliconazole: 0.0002-0.002µg/ml, terbinafine: 0.008-0.125µg/ml, efinaconazole: 0.008-0.125µg/ml, ciclopirox olamine: 0.03-0.5µg/ml, itraconazole: 0.06-2µg/ml, griseofulvin: 0.25-4µg/ml, amorolfine hydrochloride: 0.125-4µg/ml and tavaborole: 1-16µg/ml. Luliconazole, efinaconazole and terbinafine were the most potent antifungals against Tbenhamiae complex isolates, regardless of the geographic locations where strains were isolated. These data might help dermatologists to develop effective therapies for successful treatment of infections due to Tbenhamiae complex species.

Ringworm in dogs

Dermatophytosis is a common self limiting zoonotic fungal disease which coused by infection of certain types of dermatophyte (1,2).that fungi of 3 fundamental genera specifically, Microsporum, Trichophyton and epidermophyton. We wont wait its self limiting cause it Causes economic losses. That disease spread fastly amoung all existing dogs and may be owners. And take an extended time in treatment. Dermatophytosis is generally called as ringworm. And that is visibly portrayed by multi-focal alopecia and hulls on the skin with a selected formation (3,4). This infection has gained specific attention publicly health, because it distributed all over the world. After touching the contaminated items or specimens, similar to hair, soil or outside on the epidermal layer of diseased animals, infection of dermatophytosis in human happen –(5). We have noted a big increase in annual cases od dermatophytosis in all species humans and animals particularly dogs&cats (6). Scientists noted that dermatophytosis's cases are more predominant in little and female cat than male cat. In Indonesia, much researches has been done on dogs shows that : in Yogyakarta, Indonesia (34%) of dogs were positive of dermatophytosis (7). While that number ranges from 20%to30% in Europe (8). The results of different investigations recognized that main species causing dermatophytosis in small animals is microsporum-canis in 81.8%to97% of total cases (1,2,6). The occurrence of zoonotic dermatophytes, in dogs&cats and human compared with different species, m. Canis was the most awesome (60%) (9). We can modulate the immune-response by giving significant level of cortisol throughout along period.

Новые виды возбудителей в этиологии дерматофитозов животных-компаньонов в Московском регионе

Dermatophytic fungi Nannizzia persicolor, Nannizzia fulva, Trichophyton benhamiae and Microsporum ferrugineum were isolated from pets (cats, dogs, guinea pig) in Moscow region. The species identification was confirmed by ITS sequencing. Their macro- and micromorphological agent of dermatophytosis in companion animals. A considerable phenotypic polymorphism among M canis isolates was revealed, thus obstructing species identification. A wide prevalence of dermatophytosis in companion animals (46%) was shown, with a predominance of the zooanthropophilic species M. canis (95%). This is the first confirmed cases of pet dermatophytosis caused by N. persicolor, N. fulva, T. benhamiae and M. ferrugineum in Russian Federation.

Transcriptome sequencing revealed the inhibitory mechanism of ketoconazole on clinical Microsporum canis.

BackgroundMicrosporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans.ObjectivesIn clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood. The antifungal mechanism of KTZ needs to be studied in detail.MethodsIn this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis. The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto.ResultsAt half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated (p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ.ConclusionsThe results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.

Assessment of the subtilisin gene profile in Trichophyton verrucosum isolated from human and animal dermatophytoses in two-stage multiplex PCR.

Keratin is a fibrous and recalcitrant structural protein and the third most abundant polymer in nature after cellulose and chitin. Subtilisin-like proteases (SUB) are a group of serine endoproteases, coded by seven genes (SUB1-7), which decompose keratin structures and have been isolated from dermatophytes. Herein, we identified the SUB genes in 30 clinical isolates of Trichophyton verrucosum obtained from human and animal dermatophytosis as well as asymptomatic animal carriers. We designed and proposed a two-stage multiplex PCR technique to detect all seven genes encoding serine proteases in dermatophytes. The analysis revealed the presence SUB1 and SUB2 amplicons in all strains regardless of the host. In the group of isolates obtained from humans, all seven subtilisin genes were shown in 40% of the strains. In T. verrucosum from asymptomatic animals, none of the isolates showed the presence of all seven subtilisin genes, and only 30% had six genes. In turn, 10% of the isolates from symptomatic animals demonstrated all seven subtilisins amplicons. In conclusion, the severity of infection and ability of T. verrucosum to cause dermatophytosis in humans may not be related to specific genes but their accumulation and synergistic effects of their products. Dermatophytes are pathogenic filamentous fungi with capacity to attack keratinized structures such as skin, hair and nails, causing cutaneous superficial infections. Indeed, a biological characteristic of dermatophytes is their ability to invade keratin-rich tissues by producing enzymes. Various degrees of inflammatory responses can be induced exactly by the enzymes. Subtilisin-like proteases are endoproteases, which decompose keratin structures. Our study identifies SUB genes in clinical isolates of T. verrucosum obtained from human and animal dermatophytosis as well as asymptomatic animal carriers.

Prevalence of dermatophytosis in animals and antifungal susceptibility testing of isolated Trichophyton and Microsporum species.

Dermatophytosis is a frequently encountered dermatological problem in domestic and pet animals which negatively affects animal health and production. This study investigated the prevalence of dermatophytosis in different animals and carried antifungal susceptibility testing in the northern Indian state of Uttarakhand. A total of 157 clinical samples were collected from the animals irrespective of sex and age during a time period of 3years from December 2016 to December 2019. Direct microscopy, fungal culture, isolation, identification, and antifungal susceptibility testing were carried out. Direct microscopy and in vitro culture were equally efficient in diagnosis of dermatophytosis. The prevalence of dermatophytes ranged between 45 and 53.3% and Trichophyton mentagrophytes was the most abundant species (38%) followed by Microsporum canis (21.5%). All the species observed were isolated from goats and dogs except T. tonsurans and M. nanum, respectively. All the isolated dermatophytes were urease test positive except T. rubrum and T. verrucosum. The hair perforation test was negative for M. audouinii, along with T. rubrum and T. verrucosum, whereas only M. canis was positive for rice grain test. Only T. mentagrophytes, T. rubrum, T. verrucosum, and T. tonsurans were able to show optimum growth at 37°C. Maximum inhibition of fungal growth was exhibited by clotrimazole and least by fluconazole.