New insights on the use of bill sheath as a biomonitoring tool for mercury in two kingfisher species: A comparison with different tissues.

Bird species have been widely used as suitable bioindicators of environmental mercury (Hg). However, there is still some debate about the most suitable tissue to indicate Hg body burden in birds. For a long time, blood and feathers have proved to be relevant to monitor Hg at different time scales, and recently, bill sheath has been suggested as a potential tissue to this end. In the present study, we evaluated THg in muscle, liver, feathers, claws, and bill sheath in two waterbird species (i.e. the ringed and the Amazon kingfishers) from the Teles Pires, Juruena and Paraguay rivers. Considering all species and sites, feathers (5.47 ± 2.15 µg/g) and bill sheath (3.39 ± 1.37 µg/g) had mean THg concentrations about 2-, 3- and 10-times higher than claws, liver and muscle, respectively. When bird species were segregated, the ringed kingfisher showed THg values 1.8 times higher than the Amazon kingfisher in all tissues. Moreover, results showed that the Amazon kingfisher from the Juruena and Teles Pires rivers was clearly separated from the Paraguay River (control site), and was associated with higher THg values in the claws and feathers. Results obtained for the THg concentrations in bill sheath, muscle and liver tissues of the Amazon kingfisher using multivariate analysis of canonical variates (CVA) showed a pattern of segregation between the sampling areas, being the highest THg values in Teles Pires River samples. The largest bill sheath vector in the CVA suggests that this tissue is a key variable in the segregation of the samples. Overall, feathers may be useful for effects monitoring or spatial patterns, whereas bill sheath, which are more invasive, may be advantejous for temporal trends and retrospective studies of Hg pollution.

Sporothrix spp. Biofilms Impact in the Zoonotic Transmission Route: Feline Claws Associated Biofilms, Itraconazole Tolerance, and Potential Repurposing for Miltefosine.

Sporotrichosis is the most prevalent subcutaneous mycosis globally, and it is typically caused by direct inoculation of the soil saprophytic fungus Sporothrix spp. into the patients' skin. However, sporotrichosis has an important zoonotic transmission route between cats and humans in hot-spot endemic areas such as Brazil. Antifungal itraconazole is the first-line treatment; however, it is frequently associated with recurrence after withdrawal, mainly on cats. Biofilms are important resistance structures related to the environmental persistence of most microorganisms. In the present work, we evaluated Sporothrix yeasts' ability to form biofilms in an ex vivo model of infected claws of cats. Using scanning electron microscopy, we demonstrated the presence of fungal biofilms in the claws of cats diagnosed with sporotrichosis confirmed by isolation of Sporothrix spp. in culture. We present here evidence of antibiofilm activity of miltefosine and suggest its use off-label as an antifungal as a putative alternative to itraconazole against Sporothrix biofilms. Claw contamination could sustain infections through a continuous inoculation cycle between open lesions and cat claws. Our results further support the off-label use of miltefosine as a promising alternative, especially for mycosis refractory to conventional treatment.

Effect of trimming of overgrown and deformed claws in goats on morphometric measurements.

Reduced welfare and productivity of dairy goats have often been associated with poor claw health, especially conditions such as claw overgrowth and deformations. It is known that periodic claw trimmings have prophylactic and therapeutic effects on these problems, and this study aimed to evaluate if the additional use of an angle grinder to finish trimming overgrown and deformed goat claws, after the usual trimming using hoof shears, could provide further changes in these claws. For this, twelve Saanen goats (57.29 ± 11.15 kg of body weight, 3.08 ± 1.78 years old) were selected by presence of severe claw overgrowth, and absence of claw alterations of other nature. Their claws were trimmed in two steps, first using hoof shears and then using an angle grinder. Morphometric, baropodometric, and conformational aspects of all claws were assessed before claw trimming and after each trimming step. To analyse the effects of the trimming steps in each claw, the Tukey's test was used on parametric data, with 5% probability, and descriptive statistics were used on non-parametric data. Although this is a small pilot study, results suggest that using an angle grinder after the use of hoof shears, could further reduce heel length and sole width of claws, as well as reduce the number of deformed claws. The incorporation of the second trimming tool, could also further increase the frequency with which the point of maximum pressure was found in the toes, rather then in the heels of the claws as seen in deformed claws.

Biofilm formation on cat claws by Sporothrix species: An ex vivo model.

This work aimed to evaluate the ability of Sporothrix species to attach and form biofilm on the surface of cat claws as an ex vivo model. A total of 14 strains (5 Sporothrix brasiliensis, 3 Sporothrix schenckii s. str., 3 Sporothrix globosa and 3 Sporothrix mexicana) were used. The biofilms were incubated for periods of 01, 03, 07, 10 and fifteenth 15 days. Their metabolic activities were evaluated by the XTT reduction assay and the morphology and structure were investigated by scanning electron microscopy (SEM). The analysis of the SEM images revealed that all the species can form biofilms on cat claws. The metabolic activity in the ex vivo biofilms was similar to that found in in vitro biofilms when incubated for the same period. This is the first report of an ex vivo biofilm model involving cat claws. The ability to form biofilms on cat claws can increase the viable period of the fungus and consequently the number of possibly infected animals and people.

Imprint and spreading techniques for the isolation and identification of subungueal fungi in claws of healthy cats / Técnicas de imprint e espalhamento para isolamento e identificação de fungos subungueais em garras de gatos saudáveis

The aim of this study was to compare imprint and spreading techniques for the isolation and identification of colonies of pathogenic and non-pathogenic fungus in the claws of semidomiciliated cats. We evaluated 150 felines, subdivided into three groups. In the first and second groups, the animals were submitted to the imprint technique in Petri dishes containing Selective Mycobiotic Agar: 50 animals underwent antisepsis of the claws of the thoracic limbs and 50 underwent antisepsis of the claws of only one of the thoracic limbs. The third group (50 animals) was submitted to the spreading technique, whose material was collected by rubbing a sterile swab moistened with brain-heart infusion broth, in the claws of the forelimbs, where an aliquot of the material was transferred to Petri dishes containing Selective Mycobiotic Agar. The material was stored at 25ºC for 30 days. The readings were performed on days 05, 07, 15, and 30 post incubation. Using the imprint technique performed under the conditions of this experiment, we were not able to isolate and identify the colonies because since day 05, they were overlapped, making isolation and subsequent identification impossible. From the spreading technique, Mucor sp. (54,34%), Rhodotorula sp. (28,26%), Fusarium sp. (21,73%), Aspergillus sp. (21,73%), Trichoderma sp. (19,56%), Penicillium sp. (19,56%), Cladosporium sp. (10,86%), Rhizo

O objetivo deste estudo foi comparar as técnicas de imprint e espalhamento para o isolamento e identificação de colônias de fungos patogênicos e não patogênicos em garras de gatos semidomiciliados. Para tanto, foram avaliados 150 gatos, subdivididos em três grupos de 50 animais. No primeiro e segundo grupos, os gatos foram submetidos à técnica de imprint em placas de Petri contendo Agar Micobiótico Seletivo. No primeiro grupo, os gatos foram submetidos à antissepsia com eta-nol 70% das garras dos membros torácicos e no segundo grupo os animais foram submetidos à antissepsia com etanol 70% das garras de apenas um dos membros torácicos. O terceiro grupo foi submetido à técnica de espalhamento, cujo material foi coletado esfregando-se um swab estéril umedecido em caldo infusão cérebro-coração nas garras dos membros anteriores, a par-tir do qual uma alíquota do material foi transferida para placas de Petri contendo Ágar Micobiótico Seletivo. O material foi armazenado a 25 ° C por 30 dias. As leituras foram realizadas nos dias 5, 7, 15 e 30 após a incubação. Utilizando a técnica de imprint realizada nas condições deste experimento, não fomos capazes de isolar e identificar as colônias, uma vez que desde o dia 5 elas estavam sobrepostas. A partir da técnica de espalhamento, Mucor sp. (54,34%), Rhodotorula sp. (28,26%), Fusarium sp. (21,73%), Aspergillus sp. (21,73%), Trichoderma sp. (19,56%), Penicillium sp. (19,56%), Cladosporium sp. (10,86%), Rhizopussp. (8,68%), Acremonium sp. (6,5%), Exophialia sp. (6,5%), Paecilomyces sp. (4,34%), Trichosporon sp. (4,34%) e Geotrichumsp. (2,17%) foram isolados. Concluiu-se que a técnica de espalhamento mostrou-se útil no isolamento de colônias de fungos em garras felinas, e os animais não apresentam sintomas, o que sinaliza a importância deles como possíveis fontes de exposição para os tutores. Os gatos foram negativos para Sporothrix sp. pelas técnicas de imprint e espalhamento.

Imprint and spreading techniques for the isolation and identification of subungueal fungi in claws of healthy cats / Técnicas de imprint e espalhamento para isolamento e identificação de fungos subungueais em garras de gatos saudáveis

The aim of this study was to compare imprint and spreading techniques for the isolation and identification of colonies of pathogenic and non-pathogenic fungus in the claws of semidomiciliated cats. We evaluated 150 felines, subdivided into three groups. In the first and second groups, the animals were submitted to the imprint technique in Petri dishes containing Selective Mycobiotic Agar: 50 animals underwent antisepsis of the claws of the thoracic limbs and 50 underwent antisepsis of the claws of only one of the thoracic limbs. The third group (50 animals) was submitted to the spreading technique, whose material was collected by rubbing a sterile swab moistened with brain-heart infusion broth, in the claws of the forelimbs, where an aliquot of the material was transferred to Petri dishes containing Selective Mycobiotic Agar. The material was stored at 25ºC for 30 days. The readings were performed on days 05, 07, 15, and 30 post incubation. Using the imprint technique performed under the conditions of this experiment, we were not able to isolate and identify the colonies because since day 05, they were overlapped, making isolation and subsequent identification impossible. From the spreading technique, Mucor sp. (54,34%), Rhodotorula sp. (28,26%), Fusarium sp. (21,73%), Aspergillus sp. (21,73%), Trichoderma sp. (19,56%), Penicillium sp. (19,56%), Cladosporium sp. (10,86%), Rhizo(AU)

O objetivo deste estudo foi comparar as técnicas de imprint e espalhamento para o isolamento e identificação de colônias de fungos patogênicos e não patogênicos em garras de gatos semidomiciliados. Para tanto, foram avaliados 150 gatos, subdivididos em três grupos de 50 animais. No primeiro e segundo grupos, os gatos foram submetidos à técnica de imprint em placas de Petri contendo Agar Micobiótico Seletivo. No primeiro grupo, os gatos foram submetidos à antissepsia com eta-nol 70% das garras dos membros torácicos e no segundo grupo os animais foram submetidos à antissepsia com etanol 70% das garras de apenas um dos membros torácicos. O terceiro grupo foi submetido à técnica de espalhamento, cujo material foi coletado esfregando-se um swab estéril umedecido em caldo infusão cérebro-coração nas garras dos membros anteriores, a par-tir do qual uma alíquota do material foi transferida para placas de Petri contendo Ágar Micobiótico Seletivo. O material foi armazenado a 25 ° C por 30 dias. As leituras foram realizadas nos dias 5, 7, 15 e 30 após a incubação. Utilizando a técnica de imprint realizada nas condições deste experimento, não fomos capazes de isolar e identificar as colônias, uma vez que desde o dia 5 elas estavam sobrepostas. A partir da técnica de espalhamento, Mucor sp. (54,34%), Rhodotorula sp. (28,26%), Fusarium sp. (21,73%), Aspergillus sp. (21,73%), Trichoderma sp. (19,56%), Penicillium sp. (19,56%), Cladosporium sp. (10,86%), Rhizopussp. (8,68%), Acremonium sp. (6,5%), Exophialia sp. (6,5%), Paecilomyces sp. (4,34%), Trichosporon sp. (4,34%) e Geotrichumsp. (2,17%) foram isolados. Concluiu-se que a técnica de espalhamento mostrou-se útil no isolamento de colônias de fungos em garras felinas, e os animais não apresentam sintomas, o que sinaliza a importância deles como possíveis fontes de exposição para os tutores. Os gatos foram negativos para Sporothrix sp. pelas técnicas de imprint e espalhamento.(AU)

Residue Depletion of Florfenicol and Florfenicol Amine in Broiler Chicken Claws and a Comparison of Their Concentrations in Edible Tissues Using LC⁻MS/MS.

Antimicrobial residues might persist in products and by-products destined for human or animal consumption. Studies exploring the depletion behavior of florfenicol residues in broiler chicken claws are scarce, even though claws can enter the food chain directly or indirectly. Hence, this study intended to assess the concentrations of florfenicol (FF) and florfenicol amine (FFA)-its active metabolite-in chicken claws from birds that were treated with a therapeutic dose of florfenicol. Furthermore, concentrations of these analytes in this matrix were compared with their concentrations in edible tissues at each sampling point. A group of 70 broiler chickens were raised under controlled conditions and used to assess residue depletion. Sampling points were on days 5, 10, 20, 25, 30, 35, and 40 after ceasing treatment, thus extending beyond the withdrawal period established for muscle tissue (30 days). Analytes were extracted using HPLC-grade water and acetone, and dichloromethane was used for the clean-up stage. Liquid chromatography coupled to mass spectroscopy detection (LC⁻MS/MS) was used to detect and quantify the analytes. The analytical methodology developed in this study was validated in-house and based on the recommendations described in the Commission Decision 2002/657/EC from the European Union. Analyte concentrations were calculated by linear regression analysis of calibration curves that were fortified using an internal standard of chloramphenicol-d5 (CAF-d5). The depletion time of FF and FFA was set at 74 days in claws, based on a 95% confidence level and using the limit of detection (LOD) as the cut-off point. Our findings show that FF and FFA can be found in chicken claws at higher concentrations than in muscle and liver samples at each sampling point.

Residue depletion of oxytetracycline (OTC) and 4-epi-oxytetracycline (4-epi-OTC) in broiler chicken's claws by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Antibiotics are widely used in poultry production for the treatment of bacterial diseases. However, residues may remain in products and by-products destined for human consumption or animal feeding. The claws of chickens, which are a by-product of the poultry industry, can directly or indirectly enter the food chain as meals destined to feed other productive animals. Thus, it becomes necessary to determine and quantify antimicrobial residues present in this matrix. The objective of the study was to assess the depletion of oxytetracycline (OTC) and its metabolite 4-epi-OTC in broiler chicken's claws. Claws of 32 broilers treated with a therapeutic dosage of 10% OTC during 7 days were analysed. Samples were taken at days 3, 9, 15 and 19 post-treatment. As for the control group, eight broiler chickens were raised under the same conditions. Extraction was carried out through EDTA-McIlvaine buffer, and clean-up employed a SPE C-18 Sep-Pak®. Instrumental analysis was performed through LC-MS/MS. The concentrations of both analytes were determined in claw samples until day 19 post-treatment. Average concentrations were within the LOD (20 µg kg-1) and LOQ (22 µg kg-1) for OTC and 84 µg kg-1 for 4-epi-OTC. Withdrawal times (WDTs) of 39 days for OTC and 54 days for 4-epi-OTC were established in claws based on 95% confidence. These findings demonstrate that claws can be a source of antimicrobial residue entry into the food chain, since the results showed that OTC and its metabolite can be found in chicken's claws for long periods, even exceeding the average lifespan of a broiler chicken.

Non-lethal sampling for mercury evaluation in crocodilians.

Mercury (Hg) is a ubiquitous environmental contaminant that poses potential threats to ecosystems due to its toxicity to humans and wildlife. The development of non-lethal sampling techniques is a critical step for evaluation of Hg in threatened species in tropical floodplain environments, where most of Hg found is the result of land use and gold mining activities, and more methylation sites are available. We evaluated the spatial and seasonal effectiveness of caudal scutes and claws to estimate Hg bioaccumulation in crocodilians (Caiman yacare), in the scarcely documented Pantanal. Hence, we investigated the potential for Hg bioaccumulation in top predators according to its proximity to mining sites, and in water bodies with different hydrological characteristics and connectivity with the main river during two phases of the flood pulse (dry and flood). The highest Hg concentrations were detected in caimans captured close to mining activities, in claws (2176 ng g(-1) ww) and caudal scutes (388 ng g(-1) ww). THg concentration in claws was related to the flood season and its mean concentration was thirteen fold higher than Hg concentration in scutes during whole year. Both tissues were found to be effective as non-lethal sampling techniques for measuring Hg bioaccumulation in reptiles over time. Nevertheless, claw tissue seems to have a more consistent result, since its constitutional chemical characteristics makes it a better indicator of spatial patterns that influence on Hg exposure.

Oniquite lupóide isolada em cão: relato de caso / Isolated lupoid onychitis in a dog: a case report

A Oniquite Lupóide (OL) é uma síndrome que afeta várias unhas e leitos ungueais, conduzindo a onicomadesia, onicodistrofia e onicalgia, sendo a presença de onicorrexis e onicosquizia variáveis. Acredita-se que sua etiologia seja idiopática, porém sua fisiopatologia pode estar relacionada a distúrbio auto-imunes, farmacodérmicos ou a hipersensibilidade a trofoalérgenos. Várias raças já foram descritas como predispostas, sendo cães jovens, de meia idade e adultos mormente acometidos, sem aparente predisposição sexual. O presente estudo descreve um caso de OL isolada em uma cadela da raça Yorkshire Terrier, de 12 anos de idade, a qual apresentava um excessivo crescimento da unha do quarto dígito do membro pélvico esquerdo, onicalgia e claudicação com evolução de seis meses

Lupoid Onychitis is a syndrome that affects several nails and nail beds, leading to onychomadesis, onychodystrophy and onicalgia, and the presence of onychorrhexis and onychoschizia are variables. It is believed that the etiology is idiopathic, but its pathophysiology may be correlated with autoimmune disorders, pharmacodermias or hypersensibility to drugs or food allergens. Several races have been described, with young, adult and middle-aged dog’s particularly affected, with no apparent sex predisposition. This study describes a case of isolated Lupoid Onychitis in a dog Yorkshire Terrier, 12 years of age, who presented an excessive growth of the nail of the fourth digit of the left pelvic limb, pain and lameness with the evolution of six months

Oniquite lupóide isolada em cão: relato de caso / Isolated lupoid onychitis in a dog: a case report

A Oniquite Lupóide (OL) é uma síndrome que afeta várias unhas e leitos ungueais, conduzindo a onicomadesia, onicodistrofia e onicalgia, sendo a presença de onicorrexis e onicosquizia variáveis. Acredita-se que sua etiologia seja idiopática, porém sua fisiopatologia pode estar relacionada a distúrbio auto-imunes, farmacodérmicos ou a hipersensibilidade a trofoalérgenos. Várias raças já foram descritas como predispostas, sendo cães jovens, de meia idade e adultos mormente acometidos, sem aparente predisposição sexual. O presente estudo descreve um caso de OL isolada em uma cadela da raça Yorkshire Terrier, de 12 anos de idade, a qual apresentava um excessivo crescimento da unha do quarto dígito do membro pélvico esquerdo, onicalgia e claudicação com evolução de seis meses(AU)

Lupoid Onychitis is a syndrome that affects several nails and nail beds, leading to onychomadesis, onychodystrophy and onicalgia, and the presence of onychorrhexis and onychoschizia are variables. It is believed that the etiology is idiopathic, but its pathophysiology may be correlated with autoimmune disorders, pharmacodermias or hypersensibility to drugs or food allergens. Several races have been described, with young, adult and middle-aged dogs particularly affected, with no apparent sex predisposition. This study describes a case of isolated Lupoid Onychitis in a dog Yorkshire Terrier, 12 years of age, who presented an excessive growth of the nail of the fourth digit of the left pelvic limb, pain and lameness with the evolution of six months(AU)