COX-2 Is Downregulated in Human Stenotic Aortic Valves and Its Inhibition Promotes Dystrophic Calcification.

Calcific aortic valve disease (CAVD) is the result of maladaptive fibrocalcific processes leading to a progressive thickening and stiffening of aortic valve (AV) leaflets. CAVD is the most common cause of aortic stenosis (AS). At present, there is no effective pharmacotherapy in reducing CAVD progression; when CAVD becomes symptomatic it can only be treated with valve replacement. Inflammation has a key role in AV pathological remodeling; hence, anti-inflammatory therapy has been proposed as a strategy to prevent CAVD. Cyclooxygenase 2 (COX-2) is a key mediator of the inflammation and it is the target of widely used anti-inflammatory drugs. COX-2-inhibitor celecoxib was initially shown to reduce AV calcification in a murine model. However, in contrast to these findings, a recent retrospective clinical analysis found an association between AS and celecoxib use. In the present study, we investigated whether variations in COX-2 expression levels in human AVs may be linked to CAVD. We extracted total RNA from surgically explanted AVs from patients without CAVD or with CAVD. We found that COX-2 mRNA was higher in non-calcific AVs compared to calcific AVs (0.013 ± 0.002 vs. 0.006 ± 0.0004; p < 0.0001). Moreover, we isolated human aortic valve interstitial cells (AVICs) from AVs and found that COX-2 expression is decreased in AVICs from calcific valves compared to AVICs from non-calcific AVs. Furthermore, we observed that COX-2 inhibition with celecoxib induces AVICs trans-differentiation towards a myofibroblast phenotype, and increases the levels of TGF-ß-induced apoptosis, both processes able to promote the formation of calcific nodules. We conclude that reduced COX-2 expression is a characteristic of human AVICs prone to calcification and that COX-2 inhibition may promote aortic valve calcification. Our findings support the notion that celecoxib may facilitate CAVD progression.

Detection of selected pathogens in Apennine wolf (Canis lupus italicus) by a non-invasive GPS-based telemetry sampling of two packs from Majella National Park, Italy.

In this study, a multi-pathogens survey was conducted to verify the sanitary status of two Italian wolf packs of Majella National Park. Twenty fecal samples (10/pack) were collected using a sampling protocol, based on the combining data from radio-collared wolves with geographic information system (GIS) analysis, allowing to mark off the home range of packs and to recover group-specific and high-quality specimens. Virological screening against the most prevalent canine viruses (protoparvovirus, distemper virus, adenoviruses, and coronaviruses) was carried out by molecular methods, while parasites were detected by means of copromicroscopic and molecular analysis. Canine parvovirus type 2b (CPV-2b) is the most prevalent virus in both packs (7/20), followed by canine adenovirus type 2 (CAdV-2), while no sequences of canine distemper virus and coronaviruses were detected. The sequence analysis of the viruses demonstrated the domestic origin of the infection, highlighting the importance of vaccination of local dogs in order to reduce the risk of exposure of wildlife to these pathogens. Fourteen samples resulted positive for parasites. Capillaria aerophila (sin. Eucoleus aerophilus), Ancylostoma/Uncinaria, Trichuris vulpis eggs, Sarcocystis spp., Cystoisospora canis, and Angiostrongylus vasorum larvae were identified. Echinococcus granulosus sensu stricto (ovine genotype G1) and Giardia duodenalis(canid-specific Assemblage C) were also characterized, providing insights into the wolves' diet and their effects on environmental contamination. The sampling protocol applied in this study, based on a multidisciplinary approach, represents an innovative tool for the survey of Apennine wolf, able to integrate sanitary data with the ecological and demographic features of this population.

Whole Genome Sequencing of Escherichia coli and Enterococcus spp. in wildlife-livestock interface: a pilot study.

OBJECTIVES: This pilot study provides a multidisciplinary investigation to monitor livestock-wildlife interface. Ecological data, microbiological investigations, and whole genome sequencing were used to characterize eight bacterial isolates obtained from sympatric domestic and wild ruminants in Maiella National Park (Italy) in terms of genetic patterns of antimicrobial resistance. METHODS: Using selective culturing of fresh fecal samples of monitored and georeferenced populations of Apennine chamois, goats, red deer, and sheep, Escherichia coli, Enterococcus faecium, and Enterococcus faecalis isolates were isolated and subjected to minimum inhibitory concentration determination and whole genome sequencing. RESULTS: The analyzed isolates showed phenotypic and genotypic resistance to tetracycline and critically important antibiotics such as linezolid and carbapenems. Virulence genes related to biofilm regulation and Shiga toxins were also detected. Furthermore, serotypes related to nosocomial infections, harbouring plasmids recognized as important mobile resistance gene transmitters, were identified. CONCLUSIONS: This multidisciplinary pilot study represents a promising initial step to identify the environmental drivers and the transmission routes of antimicrobial resistance and virulence factors, providing new data on bacteria from rare and endangered species such as Apennine chamois.

Antibiotic Resistance in the Apennine Wolf (Canis lupus italicus): Implications for Wildlife and Human Health.

The Apennine wolf (Canis lupus italicus) is a subspecies of gray wolf that is widespread throughout Italy. Due to hunting and habitat loss, their population declined dramatically in the late 19th and early 20th centuries, but conservation efforts improved to restore the species to an estimated population of 3300 individuals. The presence of antibiotic-resistant bacteria in Apennine Wolf may pose a risk to its health and survival, as well as the health of other animals in its environment. In this study, we investigated the antibiotic resistance profiles of bacteria collected from Apennine wolves admitted to the Wildlife Research Center of Maiella National Park (Italy) in 2022. A total of 12 bacteria collected from four wolves were isolated and tested for susceptibility to antibiotics used in veterinary medicine and to critically important antibiotics for human health by means of the Vitek 2 system. All isolates were resistant to at least one antibiotic, and six bacteria were multidrug resistant to critically important antibiotics (third-generation cephalosporins, carbapenems and fluoroquinolones). The results of this pilot study have allowed for the characterization of resistant profiles in Escherichia coli, Enterococcus faecalis and other bacterial species not previously reported in Apennine wolves. Our findings provide important insights into antibiotic resistance in wildlife and its potential implications for the conservation of biodiversity and public health.

Antibiotic-Resistant Bacteria Dissemination in the Wildlife, Livestock, and Water of Maiella National Park, Italy.

Antimicrobial resistance (AMR) is a global health concern that has been linked to humans, animals, and the environment. The One Health approach highlights the connection between humans, animals, and the environment and suggests that a multidisciplinary approached be used in studies investigating AMR. The present study was carried out to identify and characterize the antimicrobial resistance profiles of bacteria isolated from wildlife and livestock feces as well as from surface water samples in Maiella National Park, Italy. Ecological and georeferenced data were used to select two sampling locations, one where wildlife was caught within livestock grazing areas (sympatric group) and one where wildlife was caught outside of livestock grazing areas (non-sympatric group). Ninety-nine bacterial isolates from 132 feces samples and seven isolates from five water samples were collected between October and December 2019. The specimens were examined for species identification, antibiotic susceptibility and molecular detection of antibiotic resistance. Forty isolates were identified as Escherichia coli, forty-eight as Enterococcus spp., eight as Streptococcus spp. and ten as other gram-negative bacteria. Phenotypic antibiotic resistance to at least one antimicrobial agent, including some antibiotics that play a critical role in human medicine, was detected in 36/106 (33.9%, 95% CI: 25-43) isolates and multidrug resistance was detected in 9/106 isolates (8.49%, 95% CI: 3.9-15.5). In addition, genes associated with antibiotic resistance were identified in 61/106 (57.55%, 95% CI: 47.5-67) isolates. The samples from sympatric areas were 2.11 (95% CI: 1.2-3.5) times more likely to contain resistant bacterial isolates than the samples from non-sympatric areas. These data suggest that drug resistant bacteria may be transmitted in areas where wildlife and livestock cohabitate. This emphasizes the need for further investigations focusing on the interactions between humans, wildlife, and the environment, the results of which can aid in the early detection of emerging AMR profiles and possible transmission routes.

Welfare and Clinical Assessment on Physical Captures Followed by Anesthesia in Apennine Chamois (Rupicapra pyrenaica ornata).

The Apennine chamois (Rupicapra pyrenaica ornata) is one of the rarest subspecies in Italy, listed in Annexes II and IV of the Habitats Directive and currently included as a vulnerable subspecies in the International Union for Conservation of Nature (IUCN) Red List. The Maiella National Park population has recently been defined as a source population for reintroduction into other parks. Since collective captures allow for better selection of target animals for the establishment of new colonies, the aim of this study is to evaluate the physiological conditions and animal welfare in free-ranging Apennine chamois after collective physical capture followed by chemical immobilization with medetomidine 0.054 mg ± 0.007, ketamine 2.14 mg ± 0.28, and acepromazine 0.043 mg ± 0.006. Twenty-one Apennine chamois (18 females and 3 males) were captured and translocated for conservation purposes. The effects of capture and anesthesia were evaluated using clinical variables, hematology, serum biochemistry, and venous blood gas analysis, the latter being used in the field for the first time in chamois capture. A risk of metabolic acidosis and capture myopathy was identified, although it did not compromise the survival of 19 chamois, which adapted to novel environments and founded new colonies, as verified through GPS radiocollars. The protocol applied in this study represents an innovative approach to assessing animal physiology and welfare in collective mountain ungulate captures, useful for improving management activities for conservation purposes.

Microbial community and antimicrobial resistance in fecal samples from wild and domestic ruminants in Maiella National Park, Italy.

This study aimed to provide new insights about antimicrobial resistance genes abundance and microbial communities of wild and domestic ruminants in wildlife-livestock interface. In total, 88 fecal samples were recovered from Apennine chamois, red deer, goat, cattle and sheep, and were collected in pools. The populations under study were selected based on ecological data useful to define sympatric and non-sympatric populations. Samples were screened for commonly used in farms under study or critically important antimicrobial resistance genes (aadA2, TetA, TetB, TetK, TetM, mcr-1). The microbial community composition was found to be different based on the species and land use of animals under study. Indeed, it was mostly characterized by phyla Firmicutes in bovine, Bacteroidota in chamois and Proteobacteria in red deer. Additionally, positive correlations between antibiotic resistance genes and microbial taxa (e.g., Tet genes correlated with Firmicutes and Patescibacteria) were described. Of the antimicrobials investigated, the abundance of mcr-1 gene suggests the importance of monitoring the wildlife in order to detect the emerging resistance genes contamination in environment. This study provides new data that highlight the importance of multidisciplinary and uncultured study in order to describe the spreading of antimicrobial resistance and related contamination in the environment.

Evidence of Linezolid Resistance and Virulence Factors in Enterococcus spp. Isolates from Wild and Domestic Ruminants, Italy.

The aim of this study was to evaluate the resistance patterns against selected critically and highly important antibiotics (quinupristin/dalfopristin, vancomycin, and linezolid) in 48 Enterococcus isolates obtained from wild (red deer and Apennine chamois) and domestic (cattle, sheep, and goats) ruminants living with varying degrees of sympatry in the protected area of Maiella National Park (central Italy). According to CLSI breakpoints, 9 out of 48 isolates (18.8%) showed resistance to at least one antibiotic. One Apennine chamois isolate was resistant to all tested antibiotics. The PCR screening of related resistance genes highlighted the occurrence of msrC or cfrD in seven Enterococcus resistant isolates. In addition, msrC and vanC genes were amplified in susceptible isolates. Specific sequences of virulence genes (gelE, ace, efa, asa1, and esp) related to pathogenic enterococci in humans were amplified in 21/48 isolates (43.75%), belonging mostly to wild animals (15/21; 71.42%). This is the first report of linezolid-resistant enterococci harboring virulence genes in Italian wildlife with special regard to the red deer and Apennine chamois species. The results allow us to evaluate the potential role of wild animals as indicators of antibiotic resistance in environments with different levels of anthropic pressure.

Resistance Patterns, mcr-4 and OXA-48 Genes, and Virulence Factors of Escherichia coli from Apennine Chamois Living in Sympatry with Domestic Species, Italy.

The aim of this study was to determine and characterize potential resistance mechanisms against selected Critically Important Antibiotics in Escherichia coli isolates collected from wild and domestic ruminants living in the Maiella National Park, in Central Italy. A total of 38 isolates were obtained from red deer, Apennine chamois, cattle, sheep, and goats grazing in lands with different levels of anthropic pressure. Antimicrobial susceptibility was determined by Minimal Inhibitory Concentration testing, showing phenotypic resistance to colistin, meropenem, or ceftazidime in 9 isolates along with one bacterial strain being resistant to three of the tested antibiotics. In addition, the biomolecular assays allowed the amplification of the genes conferring the colistin (mcr-4), the carbapenems (OXA-48), penicillins and cephalosporins (TEM, SHV, CMY-1, CMY-2) resistance. In order to describe the potential pathogenicity of isolates under study, virulence genes related to Shiga toxin-producing (STEC) and enteropathogenic (EPEC) pathovars were identified. This study is the first report of mcr-4 and OXA-48 genes in resistant E. coli harboring virulence genes in Italian wildlife, with special regard to Apennine chamois and red deer species. The multidisciplinary approach used in this study can improve the early detection of emerging antibiotic resistance determinants in human-animal-environment interfaces by means of wildlife monitoring.

Ectocommensal and ectoparasites in goldfish Carassius auratus (Linnaeus, 1758) in farmed in the State of São Paulo.

Concomitant infections by several parasitic genera are rare, very debilitating and often lethal to fish reared under commercial breeding conditions. Were describe a multiple and concurrent parasite infestation in cultured goldfish Carassius auratus with skin damage (nodules and/or ulceration). Fish with skin lesions underwent necropsy, and the skin and gills were scraped and examined. Histopathological examination with Hematoxylin-Eosin and Giemsa stain, and an ultrastructure study using transmission and scanning electron microscopy were conducted. In the skin, we identified multiple-parasite infestations by Gyrodactylidae, Epistylis sp., Trichodina sp., Ichthyophthirius multifiliis, Tetrahymena sp. and Ichthyobodo necator,associated with epithelial cell hyperplasia and epidermal sloughing. Although no gross lesions were observed, were identified a large number of parasites in the gills (Epistylis sp., Piscinoodinium sp., Ichthyophthirius multifiliis, Trichodina sp., Apiosoma sp., Hexamitasp. and cysts of a trematode digenean). The observed trematodes were not identified. The associated microscopic lesions were epithelial hypertrophic and hyperplasic and exhibited fusion of secondary lamellae and epithelial cell detachment. This is the first description of a protozoan Vorticella sp. parasitizing goldfish in Brazil. Multiple ectoparasitism by protozoa and Platyhelminthes, with or without apparent tissue damage, can be fatal for goldfish raised on farms with poor management.

Effect of aminoethylcysteine ketimine decarboxylated dimer, a natural sulfur compound present in human plasma, on tert-butyl hydroperoxide-induced oxidative stress in human monocytic U937 cells.

Aminoethylcysteine ketimine decarboxylated dimer (AECK-DD) is a natural sulphur compound present in human plasma and urine and in mammalian brain. Recently, it has been detected in many common dietary vegetables. The aim of the present study was to evaluate the ability of AECK-DD to affect cellular response of U937 human monocytic cells to tert-butyl hydroperoxide-induced oxidative stress. AECK-DD was incorporated into cells, as confirmed by GC-MS analyses, without any cytotoxic effect. A 24 h treatment with 50 and 250 microM AECK-DD resulted in the incorporation of 0.10 +/- 0.01 and 0.47 +/- 0.08ng AECK-DD x 10(6) cells, respectively. U937 cells pretreated with AECK-DD (in the range 4-100 microM) showed an increased resistance to tert-butyl hydroperoxide-induced necrotic death, as revealed by a higher percent of survival measured at all incubation times with respect to control cells. Moreover, the protective effect exhibited by AECK-DD is significantly stronger with respect to that obtained with other common antioxidants (N-acetyl cysteine and trolox) and comparable, although somewhat higher, to that of vitamin E. This effect seems to be due to the ability of AECK-DD to reduce glutathione depletion and to inhibit lipid peroxidation during tert-butyl hydroperoxide treatment. It can be concluded that AECK-DD protects cultured human monocytic cells against tert-butyl hydroperoxide-induced oxidative stress and subsequent cell death, likely through an antioxidant action inside the cell. Due to its presence in both human plasma and urine, AECK-DD may play a role in the modulation of oxidative processes in vivo.

Identification of aminoethylcysteine ketimine decarboxylated dimer, a natural antioxidant, in dietary vegetables.

Aminoethylcysteine ketimine decarboxylated dimer (simply named dimer) is a natural sulfur-containing tricyclic compound detected, until now, in human urine, bovine cerebellum, and human plasma. Recently, the antioxidant properties of this compound have been demonstrated. In this investigation, the presence of aminoethylcysteine ketimine decarboxylated dimer was identified in garlic, spinach, tomato, asparagus, aubergine, onion, pepper, and courgette. Identification of this compound in dietary vegetables was performed using gas chromatography, high-performance liquid chromatography, and gas chromatography-mass spectrometry. Results from GC analysis range in the order of 10(-4) micromol of dimer/g for all the tested vegetables. These results and the lack of a demonstrated biosynthetic pathway in humans might account for a dietary supply of this molecule.

First record of Tuxophorus caligodes (Siphonostomatoida, Tuxophoridae) in sea-farmed cobia, Rachycentron canadum, in Brazil.

The cobia is the only representative of the Rachycentridae family and, because of its zootechnical qualities, production of this fish has been implemented in several countries, such as the United States, Mexico and Brazil. Tuxophorus caligodes is a widespread parasite of marine fish worldwide. For the present report, 15 juvenile cobias were collected from net cages on a fish farm in Ilhabela, state of São Paulo, Brazil, in the winter of 2011. The fish were sacrificed by means of cerebral concussion, and then weighed (280 ± 70.5 g) and measured (27 ± 1.97 cm). After external examination under a stereomicroscope, ectoparasites present on the body surface were collected, fixed and processed for identification. Out of the 15 fish examined, two were parasitized with Tuxophorus caligodes, thus indicating a prevalence of 13.3%. This is the first report of Tuxophorus caligodes in cobias in Brazil.

Ectocommensal and ectoparasites in goldfish Carassius auratus (Linnaeus, 1758) in farmed in the State of São Paulo / Ectocomensais e ectoparasitas em kinguios Carassius auratus (Linnaeus, 1758) criados em fazenda no Estado de São Paulo

Concomitant infections by several parasitic genera are rare, very debilitating and often lethal to fish reared under commercial breeding conditions. Were describe a multiple and concurrent parasite infestation in cultured goldfish Carassius auratus with skin damage (nodules and/or ulceration). Fish with skin lesions underwent necropsy, and the skin and gills were scraped and examined. Histopathological examination with Hematoxylin-Eosin and Giemsa stain, and an ultrastructure study using transmission and scanning electron microscopy were conducted. In the skin, we identified multiple-parasite infestations by Gyrodactylidae, Epistylis sp., Trichodina sp., Ichthyophthirius multifiliis, Tetrahymena sp. and Ichthyobodo necator,associated with epithelial cell hyperplasia and epidermal sloughing. Although no gross lesions were observed, were identified a large number of parasites in the gills (Epistylis sp., Piscinoodinium sp., Ichthyophthirius multifiliis, Trichodina sp., Apiosoma sp., Hexamitasp. and cysts of a trematode digenean). The observed trematodes were not identified. The associated microscopic lesions were epithelial hypertrophic and hyperplasic and exhibited fusion of secondary lamellae and epithelial cell detachment. This is the first description of a protozoan Vorticella sp. parasitizing goldfish in Brazil. Multiple ectoparasitism by protozoa and Platyhelminthes, with or without apparent tissue damage, can be fatal for goldfish raised on farms with poor management.

A infestação concomitante por vários gêneros de parasitas é uma condição rara, debilitante e letal para os peixes e está geralmente associada aos problemas ambientais e de manejo. Com o objetivo de descrever os ectoparasitos presentes em kinguios Carassius auratus com lesões cutâneas, cultivados em fazenda comercial, foram analisados 30 exemplares de peixes por esfregaços de pele lesionada e de brânquias; exame histopatológico de tecidos corados pela técnica de Hematoxilina-Eosina e Giemsa e estudo ultraestrutural por microscopia eletrônica de transmissão e varredura de tecidos coletados após necropsia. Nas lesões cutâneas, foram observados os parasitos: Gyrodactylidae, Epistylis sp., Trichodina sp., Ichthyophthirius multifiliis, Tetrahymena sp. e Ichthyobodo necator, associados à hiperplasia epitelial, descamação epidérmica e presença de infiltrado inflamatório. A infestação das brânquias, mesmo sem lesão macroscópica, evidenciou Epistylissp., Piscinoodinium sp., I. multifiliis, Trichodina sp., Apiosoma sp., Hexamita sp. e cistos de trematódeo associados à hipertrofia e hiperplasia epitelial, fusão de lamelas secundárias, descamação epitelial e presença de infiltrado inflamatório. Os trematódeos observados não foram identificados. É a primeira descrição do protozoário Vorticella sp. parasitando kinguios no Brasil. O ectoparasitismo múltiplo por protozoários e platelmintos, com ou sem lesão aparente, pode ser fatal em peixes cultivados em condições ambientais precárias.

Is hypoxia a stressor to American bullfrog tadpoles? / A hipóxia é um estressor para girinos de rã-touro?

The aim of this study was to evaluate alterations to the physiological profile of cortisol in pro-metamorphose phase tadpoles of Lithobates catesbeianus exposed to hypoxia stressor in a capture experiment and in a crowding experiment. The capture study was performed by the treatments: stress due to individual capture with a hand net, stress due to batch capture with a hand net and stress due to capture by emptying. Three simultaneous replicates was done witch 12 animals were sampled (6 normoxia - immediately blood collection) and 6 hypoxia - blood collection after 15 min of air exposition) in two collection times with 5 days by intervals. The crowding study was performed by the treatments 1 tadpole L-1, 5 tadpoles L-1 and 10 tadpoles L-1. Three simultaneous replicates was done witch 8 animals (4 normoxia and 4 hypoxia) were sampled in the zero moment (ZM) - blood collection before the experiment, 6 animals/treatment (3 normoxia and 3 hypoxia) to 4 and 8 days and 18 animals/treatment (9 normoxia and 9 hypoxia) to 12 days. The average values to plasmatic cortisol varying from 1.7 to 5.1ng mL-1 (capture study) and 1.0 to 4.2ng mL-1 (crowding study). It concludes that the biomarker tested (cortisol) showed no alterations front of the stressor used. Alternatively, a larger response pattern to these stimuli may have been expressed in another level of an unmeasured hormone (corticosterone). And the bullfrog has great ability to adapt to different management compared to other aquatic organisms, which demonstrates the plasticity of these animals.

Avaliou-se o cortisol em girinos de rã-touro (Lithobates catesbeianus), no estágio de pró-metamorfose ao mecanismo estressor de hipóxia juntamente com captura (1º experimento) e adensamento (2º experimento). O experimento de captura foi composto pelos tratamentos: Captura individual com puçá, Captura em massa com puçá e Captura por escoamento, com 3 réplicas simultâneas onde 12 organismos foram amostrados (6 normoxia - coleta imediata de sangue e 6 hipoxia - coleta de sangue após 15 min de exposição ao ar) em 2 tempos de coleta com intervalo de 5 dias. O experimento de densidade foi composto pelos tratamentos: 1 girino L-1, 5 girinos L-1 e 10 girinos L-1, com 3 réplicas simultâneas onde 8 animais (4 normoxia e 4 hipoxia) foram amostrados no momento zero (MZ) - coleta de sangue anterior ao experimento, 6 animais/tratamento (3 normoxia e 3 hipoxia) para 4 e 8 dias e 18 animais/tratamento (9 normoxia e 9 hipoxia) para 12 dias. Os valores médios para o cortisol plasmático foram de 1,7 a 5,1ng mL-1 (Experimento de Captura); e 1,0 a 4,2ng mL-1 (Experimento de Densidade). Conclui-se que o marcador biologico de estresse utilizado (cortisol) não foi alterado pelo agente extressor. Alternativamente a resposta a este estímulo pode ser expressa em outro nível hormonal (corticosterona). E a rã-touro apresenta ótima capacidade de se adaptar aos diferentes manejos se comparados a outros organismos aquáticos, o que demonstra a plasticidade destes animais.

Synthesis and characterization of a dehydrogenation product arising from the oxidation of aminoethylcysteine ketimine decarboxylated dimer.

While investigating the antioxidant properties of aminoethylcysteine ketimine decarboxylated dimer (1) (a natural substance occurring in biological fluids such as human urine and plasma and in bovine cerebellum), a previously unreported oxidation product was obtained. This compound was identified and characterized through comparison with an authentic sample prepared via Pd-catalyzed dehydrogenation of 1. This molecule is an example of an alternative oxidation pathway involving 1.

Physiological response of American bullfrog tadpoles to stressor conditions of capture and hypoxia / Resposta fisiológica de girinos de rã-touro americana submetidos aos mecanismos estressores de captura e hipóxia

The American bullfrog (Rana catesbeiana), recently named Lithobates catesbeianus is currently farmed for commercial purposes throughout various regions of Brazil. Stressful situations such as problems of management, inadequate facilities and environmental changes with consequent reduction of immunity are common in intensive production. The assessments of these situations of stress allow us detect these problems decreasing the injuries causes by confinement. The main objective of this study was to use the biological markers of plasma cortisol and glucose level and hematological parameters to evaluate the response of bullfrog tadpoles submitted to stressed mechanisms of capture and hypoxia. The animals were subjected to three treatments: stress due to individual capture with a hand net; stress due to batch capture with a hand net; and stress due to capture by emptying. The results obtained demonstrated that there were no statistically significant differences in the parameters tested when comparing the treatments with and without exposure to air (normoxia and hypoxia). Based on these results we can conclude that the stressful stimuli tested were not adequate to alter the biomarker tested. For the cortisol, probably this should have occurred due to the synergistic action between this hormone and thyroxin, which induces metamorphosis in these animals.

A rã-touro americana (Rana catesbeiana) recentemente denominada Lithobates catesbeianus é cria da com propósito comercial em várias regiões do Brasil. Situações estressantes tais como problemas de manejo, criação inadequada e alterações ambientais com consequente redução da imunidade são comuns em produções intensivas. A avaliação destas situações de estresse permite-nos detectar estes probemas e diminuir as injurias causadas pelo confinamento. O principal objetivo deste estudo foi utilizar os marcadores biológicos de cortisol, glicemia e dados hematológicos para avaliar a resposta de girinos de rã-touro submetidos aos mecanismos estressores de captura e hipóxia. Os animais foram distribuídosem três tratamentos: estresse por captura individual com puçá; estresse por captura em massa com puçá e estresse por captura por escoamento. Os resultados obtidos demostraram não haver diferenças estatisticamente significativas entre os parametros avaliados quando comparou-se os grupos com e sem exposição ao ar (normoxia e hipoxia). Com base nestes resultados pode-se concluir que os estímulos estressores avaliados não foram adequados para alterar os valores plasmáticos dos marcadores biológicos testados. Para o cortisol, isto ocorreu provavelmente em virtude da ação sinérgica deste hormônio e a tiroxina, que induz a metamorfose nestes animais.

Methylene blue photosensitized oxidation of hypotaurine in the presence of azide generates reactive nitrogen species: formation of nitrotyrosine.

In our previous study on the hypotaurine (HTAU) oxidation by methylene blue (MB) photochemically generated singlet oxygen (1O2) we found that azide, usually used as 1O2 quencher, produced, instead, an evident enhancing effect on the oxidation rate [L. Pecci, M. Costa, G. Montefoschi, A. Antonucci, D. Cavallini, Biochem. Biophys. Res. Commun. 254 (1999) 661-665]. We show here that this effect is strongly dependent on pH, with a maximum at approximately pH 5.7. When the MB photochemical system containing HTAU and azide was performed in the presence of tyrosine, 3-nitrotyrosine was produced with maximum yield at pH 5.7, suggesting that azide, by the combined action of HTAU and singlet oxygen, generates nitrogen species which contribute to tyrosine nitration. In addition to HTAU, cysteine sulfinic acid, and sulfite were found to induce the formation of 3-nitrotyrosine. No detectable tyrosine nitration was observed using taurine, the oxidation product of HTAU, or thiol compounds such as cysteine and glutathione. It is shown that during the MB photooxidation of HTAU in the presence of azide, nitrite, and nitrate are produced. Evidences are presented, indicating that nitrite represents the nitrogen species involved in the production of 3-nitrotyrosine. A possible mechanism accounting for the enhancing effect of azide on the photochemical oxidation of HTAU and the production of nitrogen species is proposed.