The antemortem diagnosis of aspergillosis in birds remains a complex challenge. A variety of diagnostic methods are currently available, including direct detection of Aspergillus components, antibody-based assays, and nonspecific markers such as serum protein electrophoresis (SPE), but their diagnostic performances remain limited. The objective of this study was to assess and compare the performances of several diagnostic approaches, specifically, galactomannan index measurement, beta-D-glucan assay, 3-hydroxybutyrate quantification, SPE, and mannoprotein detection tests. A secondary objective was to develop a predictive model of aspergillosis incorporating optimal test thresholds identified in the first phase, combined with clinical signs. A total of 101 serum and 31 plasma samples were collected from 118 aquatic bird of various species in France. Birds were classified into three categories: control individuals (n = 88), suspected aspergillosis cases (n = 7), and confirmed cases (n = 23). While individually considered tests displayed limitations in specificity, predictive modeling revealed that elevated levels of 3-hydroxybutyrate above 0.52 mmol/L (Se = 96%, Sp = 51%) combined with beta-globulins above 6.90 g/L (Se = 78%, Sp = 51%), in conjunction with concurrent respiratory clinical signs, were significantly associated with the occurrence of aspergillosis. A multivariable logistic model combining these variables achieved excellent diagnostic performance, with AUCs up to 0.98 and sensitivity above 98%. These findings suggest that these parameters, particularly when considered alongside clinical signs, may serve as more reliable indicators for detecting aspergillosis in birds.

Sarcoptes scabiei is a permanent ectoparasite causing scabies in humans and sarcoptic mange in animals, resulting in significant health and economic impacts worldwide. Current treatments exhibit limited efficacy against eggs and molting mites, leading to treatment failures. Terpenes from essential oils, including carvacrol, eugenol, and geraniol, have previously shown notable acaricidal and ovicidal activity. In this study, molting Sarcoptes mites were exposed to 0.3

Abstract Background Documented changes in spatial and seasonal tick distribution highlight the importance of continuous surveillance. This paper documents results of a year-round sampling campaign in France, including identification of ticks and associated pathogens recovered from dogs and cats, as part of the European project “Protect Our Future Too.” Methods Ticks were collected from dogs and cats presented to 35 veterinary practices from 27 administrative French departments between April 2021 and July 2022. DNA extracted from each tick sample was amplified by polymerase chain reaction (PCR) for simultaneous detection of 18 types of protozoan or bacterial microorganisms. Results Among 777 collected ticks, 6 species were morphologically identified in descending prevalence order as: Ixodes ricinus (58.3%), Dermacentor reticulatus (24.2%, mainly on dogs), I. hexagonus (7.2%, mainly on cats), Rhipicephalus sanguineus sensu lato (3.6%, mainly on dogs), I. canisuga (one tick collected from a cat), and Haemaphysalis punctata (one tick from a dog). Geographical distribution varied by tick species: I. ricinus and D. reticulatus were more frequent in northeast France, whereas R. sanguineus (s.l.) was predominant in southeast France. Ticks were collected throughout the study period but peaked in spring and early summer for I. ricinus and late winter and spring for D. reticulatus. The ticks R. sanguineus (s.l.) were collected only during summer. In total, 71.0% of the ticks were positive for DNA of at least one microorganism. Anaplasma bacteria were most frequent (up to 75.3% in I. ricinus) followed by Rickettsia (up to 49.5% in D. reticulatus). Piroplasm DNA (Babesia/Theileria/Cytauxzoon spp.) was found in 6.4% of I. ricinus, 5.3% of D. reticulatus, and 3.6% of both I. hexagonus and R. sanguineus (s.l). Borrelia burgdorferi sensu lato DNA was detected in 10.2% of I. ricinus. Mycoplasma haemominutum/haematoparvum DNA was detected in 21.8% of D. reticulatus, whereas M. turicensis DNA was detected in 14.3% of I. hexagonus. Conclusions Results of this study show that ticks are a year-round risk for dogs and cats in France, and tick-borne pathogens are present as mono- or coinfections at high frequencies. Tick control recommendations for veterinarians and dog and cat owners should incorporate these risks. Graphical abstract

Drug-resistant fungal disease must be addressed in the 2026 update to the Global Action Plan on Antimicrobial Resistance

Aspergillosis is a respiratory disease in birds, particularly falcons, often diagnosed late due to nonspecific signs and limited performance of current diagnostic tools. This study aimed to identify novel plasma biomarkers for aspergillosis comparing plasma samples from 15 healthy falcons and 15 affected falcons (9 with early and 6 with advanced aspergillosis). First, the presence of the causative agent, Aspergillus fumigatus, was ruled out through a proteotyping strategy. Proteomic profiling using data-independent acquisition mass spectrometry further identified 861 avian proteins. In advanced infection, 50 host proteins were significantly modulated in terms of abundance, including acute-phase proteins such as the haptoglobin isoform X2, alpha-1-acid glycoprotein, and serum amyloid A-like protein. Several immunoglobulin isoforms and functionally uncharacterized proteins were also differentially abundant. In contrast, no significant changes were observed between healthy and early affected birds. Comparison between early and advanced cases revealed nine proteins with significant abundance shifts, suggesting that they could be interesting disease progression markers. These findings highlight specific host protein responses in advanced aspergillosis animals and support the potential of targeted monitoring of plasma biomarkers for earlier, non-invasive diagnosis in falcons. Further validation on a larger cohort is warranted to assess clinical relevance and diagnostic performance in veterinary medicine. Significance Aspergillosis remains one of the most significant infectious diseases affecting birds, where delayed diagnosis frequently results in fatal outcomes. Current noninvasive diagnostic is still challenging due to non-specific clinical signs and limited sensitivity and specificity of current diagnostic tests. This study presents a comprehensive proteomic characterization of falcon plasma during aspergillosis, taking advantage of recent advances in high-resolution data-independent acquisition mass spectrometry and the availability of annotated falcon genomes. Comparison of healthy, early, and advanced clinical stages revealed 50 significantly dysregulated proteins, including key acute-phase proteins such as haptoglobin, serum amyloid A, alpha-1-acid glycoprotein, ceruloplasmin, and immunoglobulin light chains. These alterations reflect systemic inflammatory and immune responses to infection and highlight promising biomarkers for early, noninvasive diagnosis. This work contributes to the development of protein-based diagnostic assays, thereby improving clinical management and welfare of captive and wild birds, while also expanding proteomic resources for non-model avian species.

L’angiostrongylose est une maladie grave et potentiellement mortelle chez le chien. Le cycle évolutif du parasite responsable, Angiostrongylus vasorum , inclut des gastéropodes qui jouent le rôle d’hôtes intermédiaires. L’angiostrongylose est une maladie émergente en Europe. Les raisons de son expansion sont, en partie, liées à la dynamique des populations de renards. L’angiostrongylose est largement répandue en Europe et en France. En France, sa séroprévalence est estimée à 1,14 %. Les enquêtes auprès des vétérinaires praticiens montrent que la maladie est présente dans des foyers sur tout le territoire national et que sa prévalence semble augmenter. L’analyse des facteurs de risque permet d’identifier les animaux à risque et de mieux cibler la prévention de cette maladie. Le jeune âge et l’ingestion de limaces ou d’escargots augmentent le risque d’infestation. Un plus grand nombre de cas sont observés en automne.

This study reports the infection, diagnosis and treatment of the nematode Parafilaria bovicola in a horse (Equus caballus) in France. A soft, fluctuated and non-painful subcutaneous nodule was observed under the right eye. Ultrasound revealed a mobile verminous element in a fluid cyst. A nematode was surgically extracted from the cyst and further identified as Parafilaria bovicola by sequencing of mitochondrial cytochrome c oxidase subunit 1 gene. This first report indicates that horses can be accidental hosts of P. bovicola in enzootic areas.

Background Scabies, caused by the mite Sarcoptes scabiei, affects more than 200 million people at any one time and can be complicated by severe secondary bacterial disease. The same mite causes sarcoptic mange in companion animals, livestock and wildlife. Population-scale nuclear genomic data spanning diverse hosts and geographical regions remain limited. Methods We generated whole-genome sequence data for 106 S. scabiei samples and incorporated 19 publicly available datasets from humans, companion animals, livestock and wildlife. After quality filtering, 83 of 125 sample-level datasets were retained. We assessed ancestry, phylogenetic relationships, nucleotide diversity, genetic differentiation, gene-level variation and candidate selection signals. Results We identified 1,568,031 filter-passing nuclear variants, including 698,136 single-nucleotide polymorphisms; sample-level heterozygosity estimates ranged from 0.27% to 11%. ADMIXTURE, principal component and phylogenetic analyses supported seven ancestry clusters broadly corresponding to host-associated groups, with pairwise fixation-index estimates ranging from 0.22 to 0.92. Multi-metric analysis identified 1,600 candidate selection blocks – eight hard-sweep, 486 soft-sweep and 1,106 ancient-adaptation blocks – and six conserved low-diversity blocks. Variable genes showed nominal enrichment for DNA repair, cell adhesion and signalling, whereas invariable genes showed nominal enrichment for ribosomal function, oxidative phosphorylation and mitochondrial processes. Conclusions This expanded multi-host analysis reinforces and extends evidence that host association is a prominent correlate of genome-wide population structure in S. scabiei. The findings provide a population-genomic resource for future host-stratified One Health surveillance and investigations of transmission, adaptation and acaricide resistance in scabies and sarcoptic mange.

Veterinary infections caused by anthropophilic dermatophytes are extremely rare and, until now, have mainly involved Trichophyton rubrum. The present investigation documents 22 cases of positive culture for the anthropophilic dermatophyte species T. tonsurans in companion animals identified in France from 2021 to 2025. Trichophyton tonsurans was isolated from 13 cats and 9 dogs. Eleven animals showed symptomatic infection, whereas the others were asymptomatic carriers. Contamination most likely followed owner-to-animal transmission, suggesting a new route of spread for T. tonsurans, an increasingly reported species. Patients infected with T. tonsurans should therefore be informed of the risk of transmission to their companion animals.

The yeast Malassezia pachydermatis frequently causes otitis externa and dermatitis in dogs and cats. This often requires prolonged antifungal therapy, raising concerns about reduced susceptibility. Antifungal susceptibility testing (AFST) for Malassezia spp. remains challenging because many protocols require complex lipid supplementation and are difficult to implement in routine veterinary settings. Here, we compared three culture media for AFST of M. pachydermatis: (i) a complex, lipid-enriched RPMI formulation (RPMI1); (ii) a simplified RPMI formulation supplemented with Tween 40 and Tween 80 (RPMI2); and (iii) a Sabouraud dextrose medium supplemented with Tween 80 (SD). Forty-nine canine clinical isolates were tested against ketoconazole, itraconazole, posaconazole and terbinafine using a colorimetric broth microdilution approach and a gradient diffusion method on solid media (EzyMIC®). RPMI1 broth was unstable after freezing, resulting in weak or ambiguous resazurin readings and frequent data exclusions, and the solid formulation of RPMI1 was unsuitable for reliable gradient diffusion interpretation. By contrast, RPMI2 medium provided consistent minimal inhibitory concentration (MIC) determinations for azoles in both solid and liquid media, with comparable MIC values. Conversely, terbinafine MICs were frequently elevated in RPMI-based liquid media, often exceeding the upper testing range, whereas lower and more readily interpretable MIC values were obtained on SD agar, suggesting medium-dependent effects on terbinafine assessment. The results obtained with SD media on azoles were more heterogeneous. Overall, RPMI2 offered a practical and reproducible option for azole susceptibility testing of M. pachydermatis, while terbinafine results may require cautious interpretation depending on the testing medium.

The Trichophyton mentagrophytes complex includes pathogens of major relevance in both human and veterinary medicine. Despite growing knowledge of the rDNA Internal Transcribed Spacer (ITS) region diversity within the complex, data on circulating genotypes in animals remain scarce. We analyzed 348 T. mentagrophytes isolates obtained from companion animals in France in 2020 to describe ITS genotype distribution and assess the presence of T. indotineae and T. mentagrophytes ITS-genotype VII. Eight ITS genotypes were identified, with genotype III* largely predominant (62.4%), followed by genotypes III (22.4%) and II* (8.6%). No significant difference in genotype distribution was observed between cats and dogs. Genotype XXIV was exclusively detected in the three chinchilla isolates, while genotype XXVI-Uhrlass was identified in 17 cases. Three previously undescribed ITS genotypes were also recorded. Sequencing of tef1-α and tubb in a subset of isolates showed limited diversity within each ITS genotype, except for genotype III*, which exhibited several sequence variants. Mating-type analysis revealed both idiomorphs in the collection, although a single idiomorph was detected within each ITS genotype except genotype III*. Genotypes III and XXVI exhibited a restricted geographical distribution. Neither T. indotineae, T. mentagrophytes ITS-genotype VII, nor T. interdigitale ITS genotypes were identified. Our results highlight restricted genetic diversity among companion animal-derived isolates in France and support reduced genetic mixing among ITS genotypes, consistent with ongoing speciation processes within T. mentagrophytes species complex.

BACKGROUND:Erythemato-ceruminous otitis externa (ECOE) is commonly associated with Malassezia spp. and microbial imbalance. HYPOTHESIS/OBJECTIVE:To assess the clinical performance of an ear cleaner in dogs suffering from ECOE associated with Malassezia spp. overgrowth and to measure its impact on the microbiota. ANIMALS:Thirty privately owned dogs suffering from mild-to-moderate ECOE associated with Malassezia spp. overgrowth. MATERIALS AND METHODS:Pruritus score (PS), clinical score (0-3 Otitis Index Score [OTIS3]: based on secretions, erythema, hyperplasia and ulceration) and cytological score (CS) were assessed on Day (D)0, D7, and D14. Sterile ear swabs were used on D0 and D14 to perform fungal culture DNA extraction, and PCR amplification of the 18 ITS and 16S rRNA gene was carried out to evaluate the composition and changes of the otic microbiome. Ears were cleaned with a cerumenolytic ear cleanser daily, every other day, or twice a week depending on the secretion score. RESULTS:Fifty-seven ears (30 dogs) completed the study. PS, OTIS-3, and CS were significantly decreased at D7 and D14 (-45%, -43%, -60% and -77%, -66%, -76%, respectively; p < 0.0001 Wilcoxon test). Fungal culture was positive (Malassezia spp.) in only 34 ears at D0 and was negative in all ears at D14. Mycobiota composition was significantly different before and after treatment with an increase of fungal diversity (Shannon index; p < 0.003 Wilcoxon test) at D14 compared to D0. There was no change in bacterial composition. CONCLUSIONS AND CLINICAL RELEVANCE:In case of ECOE associated with Malassezia spp. overgrowth, the use of the tested ear cleaner showed a positive impact on the fungal dysbiosis and Malassezia spp. overgrowth.

Cutaneous leishmaniasis presents a significant challenge to public health due to its diverse clinical manifestations, resistance development, and treatment-related adverse effects. Here, we examined the efficacy of ivermectin, moxidectin (MOX), afoxolaner, and permethrin against all stages of Leishmania tropica and THP-1 cells. We also assessed the potential for resistance acquisition after 15 rounds of artificial selection. To elucidate the mode of action of MOX, we employed RNA sequencing, molecular dynamics simulation, and chloride flux assays. Additionally, we evaluated the therapeutic index of MOX using the Galleria mellonella infection model. MOX demonstrated the highest selectivity index against leishmaniasis (promastigotes: 0.58 μM; amastigotes: 0.96 μM; host cells: 60.29 μM). Moreover, MOX exhibited the lowest resistance acquisition in both promastigotes and intracellular amastigotes after 15 rounds of artificial selection, with resistance ratios of 17.23 and 4.59, respectively. Post-exposure to MOX, differential gene expression profiles showed both stage-specific and stage-unspecific enrichment of gene families involved in crucial biological processes. Moreover, molecular dynamics simulations revealed a potential neutralizing effect of MOX on the chloride channel of L. tropica. Specifically, MOX binds to the selectivity filter, potentially disrupting the osmotic equilibrium and thereby killing the parasite. The in vivo introduction of MOX significantly inhibited the growth of L. tropica in G. mellonella larvae, resulting in decreased rates of mortality and melanization. These findings indicate that MOX is a promising candidate for the treatment of L. tropica infections, warranting further investigation and potential consideration for clinical use.

Le nématode Aelurostrongylus abstrusus est l’agent responsable de l’aélurostrongylose, une parasitose respiratoire chez le chat. Le cas présenté est un chaton d’environ six mois, amené pour une dyspnée expiratoire grave et des sifflements à l’auscultation. La méthode de Baermann et une PCR ont permis de confirmer que l’agent responsable était A. abstrusus . Un traitement antiparasitaire à base de lactones macrocycliques a permis la guérison clinique et parasitaire. L’aélurostrongylose est présente sur tout le territoire français. Les enquêtes basées sur la recherche de larves L1 dans les matières fécales montrent une prévalence entre 0,7 % et 6,1 % de la population étudiée en France. Une enquête récente, basée sur un questionnaire, indique qu’un vétérinaire répondant sur cinq a déjà vu ou suspecté au moins un cas d’aélurostrongylose entre 2023 et 2024. En Europe, la séroprévalence varie de 9 % à 21,4 %. Le cycle parasitaire comprend le chat comme hôte définitif, les gastéropodes comme hôtes intermédiaires et les rongeurs comme hôtes paraténiques. Les jeunes chats qui chassent semblent particulièrement à risque d’aélurostrongylose. En France, cinq spécialités disposent d’une AMM pour le traitement ou la prévention de l’aélurostrongylose féline. Ces spécialités comportent de la moxidectine, de l’émodepside ou de l’éprinomectine.

BACKGROUND:The resistance of Aspergillus flavus to the azole antifungal drugs is an emerging problem. Mutations in the molecular targets of the azole antifungals - CYP 51 A, B and C - are possible mechanisms of resistance, but data to confirm this hypothesis are scarce. In addition, the behaviour of resistant strains in vitro and in vivo is not yet understood. OBJECTIVES:This study had 3 objectives. The first was to compare the sequences of CYP51 A, B and C in resistant and susceptible strains of A. flavus. The second was to look for the existence of a fitness cost associated with resistance. The third was to evaluate the activity of voriconazole and posaconazole on resistant strains in the Galleria mellonella model. METHODS:The CYP51 A, B and C sequences of seven resistant strains with those of four susceptible strains are compared. Fitness costs were assessed by growing the strains in RPMI medium and testing their virulence in G. mellonella larvae. In addition, G. mellonella larvae infected with strains of A. flavus were treated with voriconazole and posaconazole. RESULTS:In the CYP51A sequences, we found the A91T, C708T and A1296T nucleotide substitutions only in the resistant strains. The resistant strains showed a fitness cost with reduced in vitro growth and reduced virulence in G. mellonella. In vivo resistance to posaconazole is confirmed in a strain with the highest MIC for this antifungal agent. CONCLUSIONS:These results allow to conclude that some substitutions in CYP51 genes, in particular CYP51A, contribute to resistance to azole drugs in A. flavus. The study of the relationship between drug dosage and treatment duration with resistance and the reduction of fitness costs in resistant strains is a major perspective of this study. This work could help to establish recommendations for the treatment of infections with resistant strains of A. flavus.

Aspergillosis is a major health problem in captive penguins due to the inhalation and the development of airborne spores of opportunistic environmental molds of the genus Aspergillus. Diagnosis is often delayed and treatments, based on the use of azole antifungals, are not fully effective. This study assesses the risk of exposure to Aspergillus sp. and determines the environmental reservoirs in the direct environment of a colony of Humboldt penguins (Spheniscus humboldti) in a zoological park in Paris, and the risk of contamination with resistant isolates. Every 15 days between February and May 2022, environmental samples (air and subtract from the nests, pond water, pigeon and penguin droppings) were carried out in the penguin enclosure as well as clinical samples (one-time non-invasive sampling on chicks), and screened for Aspergillus sp. conidia. From 191 environmental samples, 264 strains of Aspergillus including 221 strains of A. fumigatus were isolated, mostly from ambient air, in the nests, and pond water. No "at risk" areas in the penguin environment have been highlighted, nor an increased risk because of the proximity with urban wild birds. However, the load of airborne Aspergillus in the nests increased significantly with outdoor temperature. Of the 221 strains isolated, we identified only one azole-resistant strain, displaying the TR34/L98H mutation in the cyp51A gene. This low prevalence of resistant strains may probably be partly explained by the urban location of the zoological park, surrounded by kilometers of urban areas without agricultural activities.

BACKGROUND:Despite changes in the epidemiology of dermatophyte infections, the incidence of fungal infections associated with Trichophyton species still remains high among dogs and cats. The objective of the present study was to isolate and characterize dermatophytes from dogs and cats in Iran. METHOD:From December 2022 to May 2023, skin and hair samples were collected from symptomatic and asymptomatic cats and dogs in Mazandaran, a northern province of Iran. The samples were then inoculated into Mycosel™ Agar. Dermatophyte isolates were identified by sequencing the internal transcribed spacer region. Antifungal susceptibility tests were conducted using the Clinical and Laboratory Standards Institute (CLSI-M38-A3). RESULT:Of the 250 samples collected (from 200 dogs and 50 cats), 20 (from 19 dogs and one cat) (8.0 %) were positive for dermatophyte growth. Based on sequence and phylogenetic analysis, all isolates belonged to T. mentagrophytes II*. Of these positive samples, 14 (70.0 %), 3 (15.0 %), 2 (10.0 %), and 1 (2.0 %) were isolated from asymptomatic stray dogs, symptomatic stray dogs, symptomatic domestic dogs, and symptomatic cats, respectively. Luliconazole and terbinafine displayed potent activity against all T. mentagrophytes isolates, with Minimum inhibitory concentration (MIC) values of 0.016 µg/ml. Miconazole and griseofulvin demonstrated higher MIC (1 and 8 µg/ml). CONCLUSION:The present study indicated that T. mentagrophytes II* asymptomatic carriage is frequent in stray dogs in Iran. The potential risk to public health needs to be evaluated However, T. mentagrophytes genotype VIII, considered as an endemic and emerging human pathogenic clone in several countries, was not detected during the present survey.

Aspergillosis is a common and deadly bird disease. Diagnosing this disease is difficult before bird death. A review of 13 studies on various birds showed that current tests have inconsistent accuracy. No test reliably diagnoses aspergillosis in birds.