Evaluation of histo-toxicity of nimesulide in black kites (Milvus migrans): a pharmacodynamic study / Avaliação da histotoxicidade da nimesulida em milhafre-preto (Milvus migrans): um estudo farmacodinâmico

The present experimental work was conducted to elucidate the toxicity of nimesulide at three different doses in black kites (Milvus migrans). M. migrans is one of the most common raptors near human habitations. The goal of the current investigation was to determine whether nimesulide is similarly hazardous to raptors as was diclofenac sodium and to investigate the acute oral toxicity of nimesulide in these birds. For this study, eight adult male black kites (M. migrans) were randomly divided into four groups. M. migrans in the control group (n = 02) were not treated with nimesulide. The other three groups were given nimesulide doses. The birds in the first (n = 02) were declared the control group. The second (n = 02), third (n = 02), and fourth groups were administered nimesulide at a low, medium, and high dose of 2, 4, and 6 mg/kg live body weight of bird/day, respectively, for 10 days. Nimesulide-addled birds became listless and despondent, then anorexic. The birds were standing there with their eyes closed and showing no signs of life. There was an increase in saliva production, a slowing of breathing, and dilated pupils. No clinical signs were observed in the control group. No mortality was seen in the control or treated groups. The control group did not show lesions of gout, but black kites intoxicated with nimesulide at 2, 4, and 6 mg/kg live body weight of bird/day showed inflammation, apoptosis, hemorrhage, necrosis, and leukocytic infiltration tissues of the liver, kidney, and heart of black kites (M. migrans) treated with different concentrations of nimesulide. The treated groups also showed apoptosis of myofibrils and hyperplasia. The hypertrophy, atrophy, fibrosis, necrosis of skeletal muscles and hemorrhage were prominent in the muscles of black kites (M. migrans) intoxicated with nimesulide. All observed histological alterations got worse in a dose-related way. There was no significant difference in AST, ALT, ALP, serum uric acid, but a significant difference was observed in the values of serum urea (p = 0.001) and serum creatinine (p = 0.019).

O presente trabalho experimental foi conduzido para elucidar a toxicidade da Nimesulida em três doses diferentes em milhafres (Milvus migrans). M. migrans é uma das aves de rapina mais comuns perto de habitações humanas. O objetivo da presente investigação foi determinar se a Nimesulida é igualmente perigosa para as aves de rapina como foi o diclofenaco sódico e investigar a toxicidade oral aguda do fármaco nessas aves. Para este estudo, 8 milhafres machos adultos (M. migrans) foram aleatoriamente divididos em 4 grupos. M. migrans no grupo controle (n = 2) não foram tratados com Nimesulida. Os outros 3 grupos receberam doses do fármaco. As aves do primeiro grupo (n = 2) foram declaradas o grupo controle. O segundo (n = 2), terceiro (n = 2) e quarto grupos receberam Nimesulida nas doses baixa, média e alta de 2, 4 e 6 mg/kg de peso corporal vivo da ave/dia, respectivamente, por 10 dias. Aves confusas com Nimesulida tornaram-se apáticas e desanimadas, depois anoréxicas. Os pássaros estavam parados com os olhos fechados e sem sinais de vida. Houve um aumento na produção de saliva, lentidão na respiração e pupilas dilatadas. Nenhum sinal clínico foi observado no grupo controle. Nenhuma mortalidade foi observada nos grupos de controle ou tratados. O grupo controle não apresentou lesões de gota, mas os milhafres intoxicados com Nimesulida nas doses de 2, 4 e 6 mg/kg peso vivo da ave/dia apresentaram inflamação, apoptose, hemorragia, necrose e infiltração leucocitária nos tecidos do fígado, rim e coração de milhafre-preto tratados com diferentes concentrações de Nimesulida. Os grupos tratados também apresentaram apoptose de miofibrilas e hiperplasia. A hipertrofia, atrofia, fibrose, necrose da musculatura esquelética e hemorragia foram proeminentes nos músculos de milhafres negros intoxicados com o fármaco. Todas as alterações histológicas observadas pioraram de forma dose-dependente. Não houve diferença significativa em AST, ALT, ALP, ácido úrico sérico, no entanto, foi observada diferença significativa nos valores de ureia sérica (p = 0,001) e creatinina sérica (p = 0,019).

Polineuropatia motora em cão adulto / Motor polyneuropathy in an adult dog

Background: Nonambulatory flaccid tetraparesis can be the result of diseases of the peripheral nervous system and it is characterized by generalized lower motor neuron (LMN) signs, as weakness, tetraparesis/tetraplegia, decreased muscle tone and reflexes. The term polyneuropathy is used for dysfunction of multiple peripheral nerves. In Brazil, there are several etiologies for polyneuropathy in dogs, such as acute idiopathic polyradiculoneuritis, botulism and myasthenia gravis. Toxoplasma gondii is an uncommon cause of LMN diseases in dogs. The aim of this report was to describe a case of flaccid tetraplegia toxoplasmosis in an adult dog with a Toxoplasma gondii serology with a markedly elevated IgG titer of 1:4096. Case: A 4-year-old intact mongrel male dog, weighing 19.6 kg, was referred to the Veterinary Medical Teaching Hospital of the Universidade Estadual de Londrina (UEL) with a 5-day history of weakness that progressed to tetraparesis. Physical examination revealed no significant changes other than the dull and unkempt coat. Neurologic examination revealed severe tetraparesis that was worse in the pelvic limbs, with decreased muscle tone in all four limbs. Postural reactions and the interdigital reflex were absent in all four limbs, as was the patellar reflex, but pain perception was present. There were no clinical signs of dysfunction on examination of the cranial nerves. Laboratory tests were performed, and creatine kinase was elevated (819 U/L). Blood was drawn to look for antibodies to Toxoplasma gondii and Neospora caninum class IgG using the indirect immunofluorescence technique. The antibody titer for Toxoplasma gondii (IgG) was 1:4096. A chest radiograph was performed to look for megaesophagus, and a pulmonary pattern suggestive of mild diffuse pneumonia was observed. Treatment was performed with sulfamethoxazole and trimethoprim, and the dog's condition improved slightly. Discussion: Based on lower motor neuron findings, the neurologic lesion was localized in the nerve roots, peripheral nerves, neuromuscular junctions, or muscles. The most important diseases in the list of differential diagnoses were immune-mediated or infectious polyradiculoneuritis (toxoplasmosis, neosporosis), myasthenia gravis, toxic polyneuropathy (botulism, chronic organophosphate poisoning), and paraneoplastic polyneuropathy. Among these differential diagnoses, polyradiculoneuritis is one of the most common. It is an idiopathic inflammatory disease. Exposure to raccoon saliva (in the U.S.), vaccination, or infection have been proposed as precipitating causes, but the triggers of this disease remain unknown. Serology for neosporosis was negative, while IgG titers for toxoplasmosis were 1:4096. In a previous study, dogs with acute polyradiculoneuritis were more likely to have T. gondii IgG serum antibody titers than dogs without neurologic signs. Infection with the protozoa T. gondii and N. caninum can cause intense polyradiculoneuritis in dogs accompanied by myositis, especially in puppies. One treatment trial was based on the administration of sulfonamide-trimethoprim with pyrimethamine, whose efficacy in the treatment of toxoplasmosis in dogs has also been reported in the literature. Neurologic deficits improved slightly, and there is a possibility that certain signs may not disappear completely because of the permanent damage caused by inflammation of the nervous system, as observed in the present case. The case had the limitation that it was not possible to perform other laboratory tests to demonstrate histopathologically the presence of Toxoplasma gondii organisms in muscles or nerves. Recovery of normal function is less likely in protozoan polyradiculoneuritis than in noninfectious polyradiculoneuritis. Thus, in the present case, the main suspicion was polyradiculoneuritis secondary to toxoplasmosis. Although it is a rare condition, it is important to consider toxoplasmosis in dogs with LMN-type tetraparesis or tetraplegia.

Preliminary analysis of reproductive, behavioral and physiological characteristics of military working dogs

The use of dogs in military work environments has always aroused great interest in the general population and determining the stress levels they go through is extremely important to maintain their welfare. The aim of this research was to evaluate if the work shifts in military working dogs leads to stress conditions and if this working influences on the reproductive performance and life quality. The study was conducted at the Military Police Kennel located at Campinas, Sao Paulo, Brazil. Eight male dogs of four different breeds (German Shepherd, Belgian Malinois Shepherd, Doberman, and Rottweiler) were evaluated during two different shifts: Working Shifts: animals working 12 hours a day with 2 hour-interval; and Control Shifts: animals that were on their day off (36 hours). Saliva samples were collected for cortisol analysis at the control and working shifts. The study was carried out over 60 days and analyzed behavior, physiology, and reproduction quality. Saliva samples, behavior observation of stereotyping, resting and moving activities and semen analysis were collected by digital stimulation (for combined second and third fractions). The salivary cortisol levels during the control and working shifts were between 0.361-0.438 and 0.312-0.592 µg/dL, respectively; the highest values were found at the end of working shifts. The animals were resting during most of the observation period, but few showed stereotypic behaviors. The testicular consistency was firm and semen parameters were within the normal values in German Shepherd, Belgian Malinois Shepherd, and Doberman dogs. However, Rottweiler dogs had a higher rate of sperm abnormalities, higher salivary cortisol levels, and more stereotypic behaviors. Nevertheless this work highlights the importance of further research relating reproduction and cortisol levels in military dogs.(AU)

Hyponatremia in sheep in Northeastern Brazil / Hiponatremia em ovinos no Nordeste do Brasil

Mineral deficiencies are important causes of productive losses in ruminant farming, especially in semiarid regions. This study aimed to determine Na, K, and Cl serum concentrations, plus the salivary concentrations of Na and K in sheep raised in a semiarid region of Northeastern Brazil, and to report the epidemiology, clinical and the pathological aspects of an outbreak of hyponatremia in sheep. For this, serum and salivary samples of 55 sheep were collected in 6 different farms in the region. Na, K, and Cl concentrations were determined by colorimetric test, using commercial kits in a semi-automatic biochemical analyzer. The mean serum concentrations ranged from 131.5±13.1mEq/L to 172.4±9.3mEq/L for Na, 6.28±1.3mEq/L to 13.9±1.4mEq/L for K, and 91.6±54.8mEq/L to 113.6±1.5mEq/L for Cl. The mean salivary concentrations ranged from 119.4±9.5mEq/L to 161.8±15.2mEq/L for Na, 10.1±3.1mEq/L to 22.3±2.3mEq/L for K, and the ratio Na:K ranged from 6.0±1.3mEq/L to 11.8±4.1mEq/L. These results revealed that most sheep raised in the farms in this region are marginally depleted in sodium. In one farm, sheep had severe clinical signs of Na deficiency consisting of haggard appearance, craving for salt, polydipsia, polyuria, dry cough, inappetence, pica, weakness, difficulty to rise and move, severe emaciation, and dehydration. On autopsy, the ruminal content was generally impacted, giving to the content an aspect of clay balls. Four sheep presented hypertrophy of the adrenal glands. Histopathologically, the width of the zona glomerulosa was expanded, there was loss of the architecture and loss of adrenocortical cell cords. The epithelial cells in the zona glomerulosa and fasciculata were pyramidal, and contained hypereosinophilic cytoplasm and hyperchromatic nuclei. These data demonstrate the occurrence of Na deficiency in different degrees in sheep farms in the State of Pernambuco and highlight the importance of providing NaCl to avoid outbreaks of hyponatremia.

Deficiências minerais são importantes causas de perdas produtivas em criações de ruminantes, sobretudo em regiões semiáridas. Este trabalho teve como objetivo determinar as concentrações séricas de Na, K e Cl, além das concentrações salivares de Na e K em ovinos criados em uma região semiárida do Nordeste do Brasil e relatar a epidemiologia e os aspectos clínicos e patológicos de um surto de hiponatremia severa em ovinos. Para isso, amostras de soro e saliva de 55 ovinos foram coletadas em 6 diferentes fazendas da região. As concentrações de Na, K e Cl foram determinadas por teste colorimétrico, utilizando kits comerciais em analisador bioquímico semiautomático. As concentrações séricas médias variaram de 131,5±13,1mEq/L a 172,4±9,3mEq/L para Na, 6,28±1,3mEq/L a 13,9±1,4mEq/L para K e 91,6±54,8mEq/L a 113,6±1,5mEq/L para Cl. As concentrações salivares médias variaram de 119,4±9,5mEq/L a 161,8±15,2mEq/L para Na, 10,1±3,1mEq/L a 22,3±2,3mEq/L para K, e a razão Na: K variou de 6,0±1,3mEq/L a 11,8±4,1mEq/L. Esses resultados revelaram que a maioria dos ovinos criados nas fazendas desta região são marginalmente sódio deficientes. Em uma fazenda, os ovinos apresentavam um quadro clínico grave de deficiência de Na. Os principais sinais clínicos consistiam em apatia, avidez por sal, polidipsia, poliúria, tosse seca, inapetência, pica, fraqueza, dificuldade de se levantar e se locomover, emagrecimento e desidratação. À necropsia, o conteúdo ruminal estava geralmente compactado, dando ao conteúdo um aspecto de bolas de argila. Quatro ovelhas apresentaram hipertrofia das glândulas adrenais. Histopatologicamente, a zona glomerulosa apresentava-se espessa, com perda da arquitetura e desarranjo dos cordões celulares. As células epiteliais da zona glomerulosa e fasciculada eram piramidais, apresentavam citoplasma hipereosinofílico e continham núcleos hipercromáticos. Esses dados demonstram a ocorrência de deficiência de Na em diferentes graus em criações de ovinos no Estado de Pernambuco e evidencia a importância do fornecimento de NaCl para a prevenção de surtos de hiponatremia.

Vector saliva controlled inflammatory response of the host may represent the Achilles heel during pathogen transmission

Infection with vector-borne pathogens starts with the inoculation of these pathogens during blood feeding. In endemic regions, the population is regularly bitten by naive vectors, implicating a permanent stimulation of the immune system by the vector saliva itself (pre-immune context). Comparatively, the number of bites received by exposed individuals from non-infected vectors is much higher than the bites from infected ones. Therefore, vector saliva and the immunological response in the skin may play an important role, so far underestimated, in the establishment of anti-pathogen immunity in endemic areas. Hence, the parasite biology and the disease pathogenesis in "saliva-primed" and "saliva-unprimed" individuals must be different. This integrated view on how the pathogen evolves within the host together with vector salivary components, which are known to be endowed with a variety of pharmacological and immunological properties, must remain the focus of any investigational study dealing with vector-borne diseases. Considering this three-way partnership, the host skin (immune system), the pathogen, and the vector saliva, the approach that consists in the validation of vector saliva as a source of molecular entities with anti-disease vaccine potential has been recently a subject of active and fruitful investigation. As an example, the vaccination with maxadilan, a potent vasodilator peptide extracted from the saliva of the sand fly Lutzomyia longipalpis, was able to protect against infection with various leishmanial parasites. More interestingly, a universal mosquito saliva vaccine that may potentially protect against a range of mosquito-borne infections including malaria, dengue, Zika, chikungunya and yellow fever. In this review, we highlight the key role played by the immunobiology of vector saliva in shaping the outcome of vector-borne diseases and discuss the value of studying diseases in the light of intimate cross talk among the pathogen, the vector saliva, and the host immune mechanisms.(AU)

Vector saliva controlled inflammatory response of the host may represent the Achilles heel during pathogen transmission

Infection with vector-borne pathogens starts with the inoculation of these pathogens during blood feeding. In endemic regions, the population is regularly bitten by naive vectors, implicating a permanent stimulation of the immune system by the vector saliva itself (pre-immune context). Comparatively, the number of bites received by exposed individuals from non-infected vectors is much higher than the bites from infected ones. Therefore, vector saliva and the immunological response in the skin may play an important role, so far underestimated, in the establishment of anti-pathogen immunity in endemic areas. Hence, the parasite biology and the disease pathogenesis in "saliva-primed" and "saliva-unprimed" individuals must be different. This integrated view on how the pathogen evolves within the host together with vector salivary components, which are known to be endowed with a variety of pharmacological and immunological properties, must remain the focus of any investigational study dealing with vector-borne diseases. Considering this three-way partnership, the host skin (immune system), the pathogen, and the vector saliva, the approach that consists in the validation of vector saliva as a source of molecular entities with anti-disease vaccine potential has been recently a subject of active and fruitful investigation. As an example, the vaccination with maxadilan, a potent vasodilator peptide extracted from the saliva of the sand fly Lutzomyia longipalpis, was able to protect against infection with various leishmanial parasites. More interestingly, a universal mosquito saliva vaccine that may potentially protect against a range of mosquito-borne infections including malaria, dengue, Zika, chikungunya and yellow fever. In this review, we highlight the key role played by the immunobiology of vector saliva in shaping the outcome of vector-borne diseases and discuss the value of studying diseases in the light of intimate cross talk among the pathogen, the vector saliva, and the host immune mechanisms.(AU)

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.(AU)

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.(AU)

Monitoramento hormonal não invasivo em aves uma revisão / Non-invasive hormonal monitoring in birds a review

Para espécies silvestres, a coleta de amostras para monitorar o estresse e a função reprodutiva sempre foi um desafio. Coletas de sangue em animais silvestres são difíceis de serem realizadas, pois sempre exige a contenção física do animal, seguida ou não de sedação ou anestesia, que acaba dificultando ou impossibilitando coletas muito frequentes, tanto pelo estresse que causa ao animal como pelos cuidados clínicos que se dever ter no uso dessas drogas. As técnicas de dosagens hormonais não invasivas vieram como uma boa opção, realizando a análise destes hormônios ou seus metabólitos. Estes são excretados nas fezes, urina, excretas, pele, pelos ou saliva, permitindo o acompanhamento por longos períodos, pois em muitos casos, não exigem manipular o animal. As amostras mais utilizadas são fezes e urina e, no caso das aves, excretas. As técnicas mais utilizadas são os ensaios imunológicos, utilizado isótopos radioativos (radioimunoensaio) ou enzimas (enzimoimunoensaio). Nesta revisão, são apresentadas as principais vantagens da dosagem não invasiva, focando principalmente seu uso em aves, utilizando excretas e abordando também as principais técnicas de dosagem.(AU)

For wild species, sampling for monitoring stress and reproductive function has always been a challenge. Blood collections in wild animals are difficult to perform because they often require physical restraint of the animal, which causes stress, or at least sedation or anesthesia, which makes it difficult or impossible for frequent collections. Noninvasive hormone monitoring techniques came as a good option, performing analysis of these hormones or their metabolites. These are excreted in feces, urine, excreta, hair or saliva, allowing the collection for long periods, since in many cases they do not require to manipulate the animal. The most commonly used samples are feces and urine, and in the case of birds, excreta. The most commonly used techniques are immunological assays, radioactive isotopes (radioimmunoassay) or enzymes (enzymeimmunoassay). In this review, the main advantages of noninvasive hormonal monitoring are presented, focusing more on their use in birds, using excreta and also addressing the main dosage techniques.(AU)

Monitoramento hormonal não invasivo em aves – uma revisão / Non-invasive hormonal monitoring in birds – a review

Para espécies silvestres, a coleta de amostras para monitorar o estresse e a função reprodutiva sempre foi um desafio. Coletas de sangue em animais silvestres são difíceis de serem realizadas, pois sempre exige a contenção física do animal, seguida ou não de sedação ou anestesia, que acaba dificultando ou impossibilitando coletas muito frequentes, tanto pelo estresse que causa ao animal como pelos cuidados clínicos que se dever ter no uso dessas drogas. As técnicas de dosagens hormonais não invasivas vieram como uma boa opção, realizando a análise destes hormônios ou seus metabólitos. Estes são excretados nas fezes, urina, excretas, pele, pelos ou saliva, permitindo o acompanhamento por longos períodos, pois em muitos casos, não exigem manipular o animal. As amostras mais utilizadas são fezes e urina e, no caso das aves, excretas. As técnicas mais utilizadas são os ensaios imunológicos, utilizado isótopos radioativos (radioimunoensaio) ou enzimas (enzimoimunoensaio). Nesta revisão, são apresentadas as principais vantagens da dosagem não invasiva, focando principalmente seu uso em aves, utilizando excretas e abordando também as principais técnicas de dosagem.

For wild species, sampling for monitoring stress and reproductive function has always been a challenge. Blood collections in wild animals are difficult to perform because they often require physical restraint of the animal, which causes stress, or at least sedation or anesthesia, which makes it difficult or impossible for frequent collections. Noninvasive hormone monitoring techniques came as a good option, performing analysis of these hormones or their metabolites. These are excreted in feces, urine, excreta, hair or saliva, allowing the collection for long periods, since in many cases they do not require to manipulate the animal. The most commonly used samples are feces and urine, and in the case of birds, excreta. The most commonly used techniques are immunological assays, radioactive isotopes (radioimmunoassay) or enzymes (enzymeimmunoassay). In this review, the main advantages of noninvasive hormonal monitoring are presented, focusing more on their use in birds, using excreta and also addressing the main dosage techniques.

Salmonelose em bezerros pelo sorovar Dublin no estado do Paraná, Brasil - aspectos clínico-patológicos / Salmonellosis in calves by serovar Dublin in Paraná State, Brazil - clinicopathological aspects

Background: Calf diarrhea remains one of the main diseases affecting the cattle industry. Persistence of this significant problem is associated with the complexity of factors that may be involved (infectious, environmental). An accurate diagnosis is essential for confirming the cause and helping clinicians and cattle producers to apply appropriate strategies in a timely manner. This report describes the histological changes according to the degree of salmonellosis severity, which is a contagious infectious disease caused by Salmonella enterica subsp. enterica, a Gram-negative bacterium, in two beef calves in northern Paraná State, southern Brazil. Cases: Two 90-day-old crossbred Angus and Nellore calves from a beef farm in northern Paraná State were referred to a Veterinary Hospital. Animal 1- developed acute clinical signs (enteritis, ataxia, and muscle rigidity) and died the day after the signs began. Gross findings included heavy and non-collapsed lungs, pulmonary oedema, hepatomegaly, enteritis, and severe diffuse typhlitis. Microscopic analysis revealed severe diffuse necrotic enteritis, typhlitis, severe diffuse interstitial pneumonia, moderate centrilobular hepatic necrosis, mild multifocal nephritis, and severe spleen and lymph node necrosis. Paratyphoid nodules were evidenced on the liver, spleen, and mesenteric lymph nodes. Animal 2- presented apathy, green fibrinous diarrhea, and died three days after the onset of clinical signs. The macroscopic examination showed moderate diffuse enteritis and hepatosplenomegaly. At the microscopic examination, many paratyphoid nodules were observed on the liver, spleen, kidneys, and Peyer's patches, which were associated with intralesional and intravascular bacilli. Additional findings included severe diffuse fibrinous necrotic enteritis with intralesional bacilli, diffuse interstitial pneumonia, multifocal splenic necrosis, lymphoid depletion, and moderate multifocal to coalescent lymphocytic nephritis. Histological Gram staining was performed on selected samples, revealing intralesional Gram-negative bacilli in the liver and intestine. Thus, liver and intestine fragments were used for the microbiological examination. Microbiological culture, isolation and biochemical tests identified the genus Salmonella spp. Then, the colonies were subjected to serological tests for serovar identification, according to ISO/TR 6579-3, which determined the serovar Dublin. After identifying the disease etiological agent, the outbreak was controlled by appropriate antibiotic therapy combined with the correction of sanitary measures. Discussion: Enteritis is a frequent disease in calves, posing a diagnostic challenge in identifying the etiological agent. In the present case, the histological, microbiological, and serological results confirmed a disseminated Salmonella spp. infection. The microscopic findings, such as interstitial pneumonia, fibrinous necrotic enteritis markedly in ileum and paratyphoid nodules in various organs, are the most common aspects of the disease. However, fibrinous cholecystitis, which is considered pathognomonic for salmonellosis, was not observed in this study. Therefore, the absence of such a lesion should not exclude the disease in sick animals. A microscopic injury score was used to determine lesion severity by assigning values from 1 to 4, wherein: 1 = no apparent lesions, 2 = mild lesions, 3 = moderate lesions, and 4 = severe lesions. Both calves were scored as 4. Multiple predisposing factors for the condition were identified in this farm such as different age animals in the same paddock and no specific paddock for sick animals, given that the infectious agent remains in feces, saliva, and nasal discharge. The serovar Dublin induces several clinical signs such as septicemic, respiratory, and enteric manifestations, making a clinical diagnosis a challenge

Saliva crystallization in sheep subjected to estrus induction and synchronization protocols

Background: The crystallization of bodily fluids, primarily saliva, has been the subject of study in many species and is asimple alternative to detect estrus because it demands neither a significant financial investment nor qualified professionalsto execute the examination. Fern pattern crystallization has been described in the cervical and nasal mucus, saliva andtear secretion, and in colostrum. Changes in salivary crystallization during the reproduction cycle are related to differenthormonal concentrations in this period. Thus, the present study has evaluated the patterns of saliva crystallization in sheepsubjected to estrus induction and synchronization protocols.Materials, Methods & Results: The sample consisted of 11 crossbreed Corriedale sheep, which were evaluated during twoexperimental periods (spring and autumn), and that underwent induction and synchronizing estrus protocols. In a randomphase of the estrus cycle (day 0), each sheep was implanted with an intravaginal device (Primer®), impregnated with 0.36g of progesterone for seven days. This device was inserted according to manufacturer’s instructions of the manufacturerand with the assistance of a specific applicator. On the day of device removal (day 7), the animals received 0.0375 mg ofD-Cloprostenol (Prolise®) and 10 mg of Folltropin® extracted from the swine pituitary (NIH-FSH-P1 of Folltropin-V) byintramuscular administration. The saliva was collected at six points during the experimental periods: day 1 (3 days beforeplacement of the implant); day 4 (day of insertion of the implant); day 9 (5 days after the insertion of the implant); day11 (day of removal of the implant and application of hormones); day 12 (24 h after removal of the implant [presumableestrus]); and day 13 (48 h after removal of the implant). Smears containing 10 µL of saliva were observed under an opticallight microscope...

Saliva crystallization in sheep subjected to estrus induction and synchronization protocols

Background: The crystallization of bodily fluids, primarily saliva, has been the subject of study in many species and is asimple alternative to detect estrus because it demands neither a significant financial investment nor qualified professionalsto execute the examination. Fern pattern crystallization has been described in the cervical and nasal mucus, saliva andtear secretion, and in colostrum. Changes in salivary crystallization during the reproduction cycle are related to differenthormonal concentrations in this period. Thus, the present study has evaluated the patterns of saliva crystallization in sheepsubjected to estrus induction and synchronization protocols.Materials, Methods & Results: The sample consisted of 11 crossbreed Corriedale sheep, which were evaluated during twoexperimental periods (spring and autumn), and that underwent induction and synchronizing estrus protocols. In a randomphase of the estrus cycle (day 0), each sheep was implanted with an intravaginal device (Primer®), impregnated with 0.36g of progesterone for seven days. This device was inserted according to manufacturers instructions of the manufacturerand with the assistance of a specific applicator. On the day of device removal (day 7), the animals received 0.0375 mg ofD-Cloprostenol (Prolise®) and 10 mg of Folltropin® extracted from the swine pituitary (NIH-FSH-P1 of Folltropin-V) byintramuscular administration. The saliva was collected at six points during the experimental periods: day 1 (3 days beforeplacement of the implant); day 4 (day of insertion of the implant); day 9 (5 days after the insertion of the implant); day11 (day of removal of the implant and application of hormones); day 12 (24 h after removal of the implant [presumableestrus]); and day 13 (48 h after removal of the implant). Smears containing 10 µL of saliva were observed under an opticallight microscope...(AU)

Alterações laboratoriais e post-mortem de um felino com doença renal crônica positivo para o vírus da imunodeficiência felina / Laboratory changes and post-death of a feline with positive chronic kidney disease for the feline immunodeficiency virus

A Imunodeficiência Viral Felina (FIV) é uma patologia infecciosa, na qual o vírus está presente na saliva de gatos infectados, o que mostra transmissão direta por ferimentos causados por mordidas, além disso, pode ocorrer transmissão vertical. A doença renal crônica (DRC)é uma das patologias mais comumente observadas em gatos infectados com FIV. O objetivo do trabalho foi descrever um relato de caso clínico de um felino com DRC portador do FIV, enfatizando suas alterações laboratoriais e post-mortem. Foi atendido no Hospital Veterinário Adrisse, Fortaleza-CE, um felino, sem raça definida, de aproximadamente sete anos. Segundo a tutora, o felino havia fugido de casa, e quando retornou, apresentava vômitos esporádicos, diarreia e anorexia há quatro dias. No exame físico, apresentava quadro de ataxia, apatia, grau de desidratação em torno de 8%, dor abdominal, mioclonias, êmese, hipotermia, nistagmo bilateral, mucosas levemente ictéricas, desnutrição, urina concentrada, úlceras na boca e hálito urêmico. Foram solicitados como exames complementares: hemograma e dosagens bioquímicas (alanina aminotransferase, creatinina, aspartatoaminotransferase), sódio, potássio, fósforo, fosfatase alcalina, cálcio, ureia, sumário de urina, ultrassonografia abdominal e snap® teste FIV/FeLV. O diagnóstico foi conclusivo através do snap FIV/FelV e a DRC se confirmou pela ultrassonografia abdominal e necropsia, através de alterações compatíveis com os sintomas e achados laboratoriais. Pode-se concluir que em felinos acometidos por FIV, são imprescindíveis a avaliação clínica e o diagnóstico de complicações renais, visto que há uma ação viral no desencadeamento de lesões renais, os quais podem ser identificados por meio de exames laboratoriais, ultrassonográficos e necroscópicos.

Feline Viral Immunodeficiency (FIV) is an infectious pathology, in which the virus is present in the saliva of infected cats, which shows direct transmission through injuries caused by bites, in addition, vertical transmission can occur. Chronic kidney disease (CKD) is one of the pathologies most commonly seen in cats infected with IVF. The objective of the study was to describe a clinical case report of a feline with CKD with IVF, emphasizing his laboratory and post-mortem changes. At the Veterinary Hospital Adrisse, Fortaleza-CE, a feline, of mixed breed, of approximately seven years was attended. According to the tutor, the cat had run away from home, and when he returned, he had sporadic vomiting, diarrhea and anorexia for four days. On physical examination, he presented ataxia, apathy, degree of dehydration around 8%, abdominal pain, myoclonus, emesis, hypothermia, bilateral nystagmus, slightly jaundiced mucous membranes, malnutrition, concentrated urine, mouth ulcers and uremic breath. Complementary tests were requested: blood count and biochemical measurements (alanine aminotransferase, creatinine, aspartate aminotransferase), sodium, potassium, phosphorus, alkaline phosphatase, calcium, urea, urine summary, abdominal ultrasound and snap® FIV/FeLV test. The diagnosis was conclusive through the FIV/FelV snap and CKD was confirmed by abdominal ultrasound and necropsy, through alterations compatible with the symptoms and laboratory findings. It can be concluded that in cats affected by FIV, clinical evaluation and diagnosis of kidney complications are essential, since there is a viral action in the triggering of kidney injuries, which can be identified through laboratory, ultrasound and necroscopic exams.

Alterações laboratoriais e post-mortem de um felino com doença renal crônica positivo para o vírus da imunodeficiência felina / Laboratory changes and post-death of a feline with positive chronic kidney disease for the feline immunodeficiency virus

A Imunodeficiência Viral Felina (FIV) é uma patologia infecciosa, na qual o vírus está presente na saliva de gatos infectados, o que mostra transmissão direta por ferimentos causados por mordidas, além disso, pode ocorrer transmissão vertical. A doença renal crônica (DRC)é uma das patologias mais comumente observadas em gatos infectados com FIV. O objetivo do trabalho foi descrever um relato de caso clínico de um felino com DRC portador do FIV, enfatizando suas alterações laboratoriais e post-mortem. Foi atendido no Hospital Veterinário Adrisse, Fortaleza-CE, um felino, sem raça definida, de aproximadamente sete anos. Segundo a tutora, o felino havia fugido de casa, e quando retornou, apresentava vômitos esporádicos, diarreia e anorexia há quatro dias. No exame físico, apresentava quadro de ataxia, apatia, grau de desidratação em torno de 8%, dor abdominal, mioclonias, êmese, hipotermia, nistagmo bilateral, mucosas levemente ictéricas, desnutrição, urina concentrada, úlceras na boca e hálito urêmico. Foram solicitados como exames complementares: hemograma e dosagens bioquímicas (alanina aminotransferase, creatinina, aspartatoaminotransferase), sódio, potássio, fósforo, fosfatase alcalina, cálcio, ureia, sumário de urina, ultrassonografia abdominal e snap® teste FIV/FeLV. O diagnóstico foi conclusivo através do snap FIV/FelV e a DRC se confirmou pela ultrassonografia abdominal e necropsia, através de alterações compatíveis com os sintomas e achados laboratoriais. Pode-se concluir que em felinos acometidos por FIV, são imprescindíveis a avaliação clínica e o diagnóstico de complicações renais, visto que há uma ação viral no desencadeamento de lesões renais, os quais podem ser identificados por meio de exames laboratoriais, ultrassonográficos e necroscópicos.(AU)

Feline Viral Immunodeficiency (FIV) is an infectious pathology, in which the virus is present in the saliva of infected cats, which shows direct transmission through injuries caused by bites, in addition, vertical transmission can occur. Chronic kidney disease (CKD) is one of the pathologies most commonly seen in cats infected with IVF. The objective of the study was to describe a clinical case report of a feline with CKD with IVF, emphasizing his laboratory and post-mortem changes. At the Veterinary Hospital Adrisse, Fortaleza-CE, a feline, of mixed breed, of approximately seven years was attended. According to the tutor, the cat had run away from home, and when he returned, he had sporadic vomiting, diarrhea and anorexia for four days. On physical examination, he presented ataxia, apathy, degree of dehydration around 8%, abdominal pain, myoclonus, emesis, hypothermia, bilateral nystagmus, slightly jaundiced mucous membranes, malnutrition, concentrated urine, mouth ulcers and uremic breath. Complementary tests were requested: blood count and biochemical measurements (alanine aminotransferase, creatinine, aspartate aminotransferase), sodium, potassium, phosphorus, alkaline phosphatase, calcium, urea, urine summary, abdominal ultrasound and snap® FIV/FeLV test. The diagnosis was conclusive through the FIV/FelV snap and CKD was confirmed by abdominal ultrasound and necropsy, through alterations compatible with the symptoms and laboratory findings. It can be concluded that in cats affected by FIV, clinical evaluation and diagnosis of kidney complications are essential, since there is a viral action in the triggering of kidney injuries, which can be identified through laboratory, ultrasound and necroscopic exams.(AU)

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...(AU)

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest / Efeito antitumoral da saliva do carrapato Amblyomma sculptum Berlese em células de neuroblastoma envolve desconstrução do citoesqueleto e parada do ciclo celular

The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (m) was slightly, but significantly (p 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and m were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.(AU)

As propriedades antitumorais de extratos de glândulas salivares de carrapatos ou proteínas recombinantes foram relatadas recentemente, mas pouco se sabe sobre as propriedades antitumorais dos componentes secretados da saliva. O objetivo deste estudo foi investigar o efeito in vitro da saliva bruta do carrapato duro Amblyomma sculptum sobre as linhagens celulares de neuroblastoma. Células SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32 e CHLA-20 foram suscetíveis à saliva, com redução de 80% na sua viabilidade em comparação com controles não tratados, como demonstrado pelo ensaio de Azul de Metileno. Investigações posteriores utilizando CHLA-20 revelaram apoptose, com aproximadamente 30% de células positivas para anexina-V, e G0/G1 (> 60%) após tratamento com saliva. O potencial de membrana mitocondrial (m) foi reduzido significativamente (p 0,05), e o citoesqueleto de actina foi desestruturado, como indicado pela microscopia de fluorescência. A viabilidade do fibroblasto humano (células HFF-1), usado como controle não tumoral, diminuiu em aproximadamente 40%. No entanto, não foram observadas alterações na progressão do ciclo celular, morfologia e m nestas células. O presente trabalho fornece novas perspectivas para a caracterização das moléculas presentes na saliva e suas propriedades antitumorais.(AU)

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest / Efeito antitumoral da saliva do carrapato Amblyomma sculptum Berlese em células de neuroblastoma envolve desconstrução do citoesqueleto e parada do ciclo celular

The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (m) was slightly, but significantly (p 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and m were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.(AU)

As propriedades antitumorais de extratos de glândulas salivares de carrapatos ou proteínas recombinantes foram relatadas recentemente, mas pouco se sabe sobre as propriedades antitumorais dos componentes secretados da saliva. O objetivo deste estudo foi investigar o efeito in vitro da saliva bruta do carrapato duro Amblyomma sculptum sobre as linhagens celulares de neuroblastoma. Células SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32 e CHLA-20 foram suscetíveis à saliva, com redução de 80% na sua viabilidade em comparação com controles não tratados, como demonstrado pelo ensaio de Azul de Metileno. Investigações posteriores utilizando CHLA-20 revelaram apoptose, com aproximadamente 30% de células positivas para anexina-V, e G0/G1 (> 60%) após tratamento com saliva. O potencial de membrana mitocondrial (m) foi reduzido significativamente (p 0,05), e o citoesqueleto de actina foi desestruturado, como indicado pela microscopia de fluorescência. A viabilidade do fibroblasto humano (células HFF-1), usado como controle não tumoral, diminuiu em aproximadamente 40%. No entanto, não foram observadas alterações na progressão do ciclo celular, morfologia e m nestas células. O presente trabalho fornece novas perspectivas para a caracterização das moléculas presentes na saliva e suas propriedades antitumorais.(AU)

Raiva canina no município de Maringá-PR, 2016 - relato de caso / Canine rabies in Maringá county, Paraná, 2016 - case report / Rabia canina en el municipio de Maringá-PR, 2016 - relato de caso

A raiva é uma antropozoonose transmitida pela inoculação do vírus contido na saliva de animais infectados. Em praticamente 100% dos casos, a raiva provoca encefalite aguda e leva seu hospedeiro ao óbito, representando um problema grave de saúde pública. Observa-se que o perfil epidemiológico da doença está se modificando ao longo dos anos, devido à transmissão significativa pelos morcegos. Relata-se um caso de raiva em um canino no município de Maringá-PR, com histórico pregresso de viagem para o estado de Macapá-AP, sem atender às medidas de prevenção à transmissão do vírus rábico.(AU)

The rabies is anthropozoonoses transmitted to humans by inoculating the vírus contained in saliva of infected animal. Causes acute encephalitis leading to death approximately 100% of cases, and this is serious health problem. It is observed that the epidemiological profile of disease is changing over the years, due the significant transmission by the bats. Is reported a rabies case, in canine, in Maringá-PR city, with previous travel history for Macapá-AP, without regard to measures to prevent the transmission of rabies virus.(AU)

La rabia es una antropozoonosis transmitida a los humanos mediante la inoculación de lo vírus contenida en la saliva del animal infectado. Causa la encefalitis aguda que conduce a la muerte en aproximadamente el 100% de los casos, y este es un problema grave de salud. Se observa que el perfil epidemiológico de la enfermedad está cambiando con los años, debido a la importante transmisión de los murciélagos. Se informó un caso de rabia, en caninos, en la ciudad de Maringá-PR, con historial de viaje anterior para Macapá-AP, sin tomar en cuenta las medidas para prevenir la transmisión del virus de la rabia.(AU)