Cryptosporidium sp. in cultivated oysters and the natural oyster stock of the state of Maranhão, Brazil / Detecção de Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão, Brasil

ABSTRACT: This study detected Cryptosporidium spp. in cultivated oysters and the natural oyster stock of the state of Maranhão and determine the elective tissue(s) to examine this protozoan. For this purpose, 200 cultivated oysters were purchased from the municipality of Raposa and another 100 from Paço do Lumiar. Additionally, 100 oysters were extracted from the natural stock of the municipality of Primeira Cruz, thus making up a total of 400 oysters. They were grouped into 80 pools consisting of 5 oysters each. From each pool, the gills and visceral mass were removed to obtain 160 pools, 80 pools for the gill group and another 80 for the visceral mass group. Then, DNA was extracted from each pool using a commercial kit with modifications. Subsequently, the protozoan DNA was detected using nested polymerase chain reaction. With this technique, the DNA of the protozoan under investigation was detected in 2.5% (n = 2/80) of the pools containing gills, with 1.25% of the pools (n = 1/80) belonging to the cultivation group of oysters and the other 1.25% (n = 1/80) to the natural stock. With the results obtained in this study, it was concluded that the analyzed oysters of the genus Crassostrea, from cultivation and natural stock groups, found in the state of Maranhão, were contaminated by Cryptosporidium spp. and may become potential sources of infection in humans and other animals. In addition, the gills are the elective tissue for the study of Cryptosporidium spp. in oysters.

RESUMO: Objetivou-se com o estudo detectar Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão e determinar o(s) tecido(s) eletivo(s) para pesquisa desse protozoário. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz, totalizando 400 ostras. Estas foram agrupadas em 80 pools constituídos por cinco animais. De cada pool, as brânquias e a massa visceral foram removidas totalizando 160 pools, sendo 80 para o grupo das brânquias e 80 para o grupo de massa visceral. Na sequência, procedeu-se à extração de DNA de cada pool com a utilização de kit comercial com modificações. Posteriormente, realizou-se a detecção do DNA do protozoário por meio da técnica de Nested-PCR. Com a técnica utilizada, foi detectado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools apenas de brânquias, sendo 1,25% pools (n=1/80) oriundos de cultivo e os outros 1,25% (n=1/80) de estoque natural. Com os resultados obtidos nesse estudo, conclui-se que as ostras analisadas do gênero Crassostrea sp., oriundas de cultivo e estoque natural no estado do Maranhão, estavam contaminadas por Cryptosporidium sp. e podem se reverter em fontes potenciais para seres humanos e outros animais. Para a pesquisa de Cryptosporidium sp. em ostras, as brânquias são o tecido eletivo.

Cryptosporidium sp. in cultivated oysters and the natural oyster stock of the state of Maranhão, Brazil / Detecção de Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão, Brasil

This study detected Cryptosporidium spp. in cultivated oysters and the natural oyster stock of the state of Maranhão and determine the elective tissue(s) to examine this protozoan. For this purpose, 200 cultivated oysters were purchased from the municipality of Raposa and another 100 from Paço do Lumiar. Additionally, 100 oysters were extracted from the natural stock of the municipality of Primeira Cruz, thus making up a total of 400 oysters. They were grouped into 80 pools consisting of 5 oysters each. From each pool, the gills and visceral mass were removed to obtain 160 pools, 80 pools for the gill group and another 80 for the visceral mass group. Then, DNA was extracted from each pool using a commercial kit with modifications. Subsequently, the protozoan DNA was detected using nested polymerase chain reaction. With this technique, the DNA of the protozoan under investigation was detected in 2.5% (n = 2/80) of the pools containing gills, with 1.25% of the pools (n = 1/80) belonging to the cultivation group of oysters and the other 1.25% (n = 1/80) to the natural stock. With the results obtained in this study, it was concluded that the analyzed oysters of the genus Crassostrea, from cultivation and natural stock groups, found in the state of Maranhão, were contaminated by Cryptosporidium spp. and may become potential sources of infection in humans and other animals. In addition, the gills are the elective tissue for the study of Cryptosporidium spp. in oysters.

Objetivou-se com o estudo detectar Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão e determinar o(s) tecido(s) eletivo(s) para pesquisa desse protozoário. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz, totalizando 400 ostras. Estas foram agrupadas em 80 pools constituídos por cinco animais. De cada pool, as brânquias e a massa visceral foram removidas totalizando 160 pools, sendo 80 para o grupo das brânquias e 80 para o grupo de massa visceral. Na sequência, procedeu-se à extração de DNA de cada pool com a utilização de kit comercial com modificações. Posteriormente, realizou-se a detecção do DNA do protozoário por meio da técnica de Nested-PCR. Com a técnica utilizada, foi detectado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools apenas de brânquias, sendo 1,25% pools (n=1/80) oriundos de cultivo e os outros 1,25% (n=1/80) de estoque natural. Com os resultados obtidos nesse estudo, conclui-se que as ostras analisadas do gênero Crassostrea sp., oriundas de cultivo e estoque natural no estado do Maranhão, estavam contaminadas por Cryptosporidium sp. e podem se reverter em fontes potenciais para seres humanos e outros animais. Para a pesquisa de Cryptosporidium sp. em ostras, as brânquias são o tecido eletivo.

Immune response dynamics of recent and chronic small ruminant lentivirus infection in the male reproductive system / Dinâmica da resposta imunológica da infecção recente e crônica por lentivírus de pequenos ruminantes no sistema reprodutor de machos

The objective of this study was to analyze the immune responses of bucks to small ruminant lentivirus (SRLV) with a focus on the reproductive system of males with recent and chronic infection. A total of 12 bucks were selected, six seronegative and six seropositive with chronic natural infection for more than 18 months (chronic infection group). After selecting the animals, the six seronegative males were intravenously inoculated with caprine arthritis-encephalitis virus (CAEV)-Co viral strain at a titer of 10-5,6 TCID50/mL. After viral inoculation, this group was called the recent infection group and was monitored weekly with the chronically infected group for 180 days with blood serum and seminal plasma Western Blot (WB) analysis. Of the animals with chronic SRLV infection, 18.94% (50/264) showed anti-SRLV antibodies in at least one of the samples, and 81.06% (214/264) were negative. Anti-SRLV antibodies were detected in 27.27% (36/132) of the blood serum samples from this group, while 10.60% (14/132) were reactive in the seminal plasma WB test. The animals inoculated with CAEV-Co became seropositive after the third week of viral inoculation. In this group, 31.06% (41/132) of seminal plasma samples had anti-SRLV antibodies, and of these, 70.73% (29/41) coincided with blood serum results. Of the remaining 29.27% (12/41), the seminal plasma sample of only three animals (RIA2, RIA3, and RIA5) had anti-SRLV antibodies. One of the animals with a recent infection presented anti-SRLV antibodies only in seminal plasma samples, possibly due to virus compartmentalization. Intermittent viral shedding was observed in both biological samples, regardless of the infection stage. The immune response in bucks with recent SRLV infection is more significant than in chronically infected animals. Regardless of the stage of infection, there is a fluctuation in antibody levels, therefore, this creates a risk of false-negative samples when performing the diagnosis.

O objetivo desse estudo foi analisar a resposta imunológica aos lentivírus de pequenos ruminantes (LVPR) com enfoque no sistema reprodutor de machos com infecção recente e crônica. Para isso, foram selecionados 12 reprodutores caprinos, sendo seis soronegativos e seis soropositivos com infecção natural crônica há mais de 18 meses (grupo com infecção crônica). Após seleção dos animais, os seis machos soronegativos foram inoculados com a cepa viral do vírus da artrite encefalite caprina (CAEV)-Co, título 10-5,6 TCID50/mL, por via intravenosa. A partir da inoculação viral este agrupamento passou a ser denominado de grupo com infecção recente e juntamente com o grupo com infecção crônica foram acompanhados, semanalmente por 180 dias, com análise dos testes de Western Blot (WB) no soro sanguíneo e plasma seminal. Nos animais com infecção crônica para LVPR, 18,94% (50/264) apresentaram anticorpos anti-LVPR em pelo menos uma das distintas amostras, e 81,06% (214/264) tiveram resultados negativos. Das amostras de soro sanguíneo do referido grupo, em 27,27% (36/132) detectou-se anticorpos anti-LVPR, enquanto que no plasma seminal 10,60% (14/132) foram reagentes no teste de WB. Nos animais inoculados com o CAEV-Co, ocorreu a soroconversão após a terceira semana da inoculação viral. Nesse grupo, 31,06% (41/132) das amostras de plasma seminal tiveram anticorpos anti-LVPR, sendo que dessas 41, 70,73% (29/41) coincidiram com resultado das amostras de soro sanguíneo. Nos 29,27% (12/41) restante, houve a detecção somente no plasma seminal e eram amostras provenientes de três animais (AIR2, AIR3 e AIR5). Em um dos animais com infecção recente, só foi identificado anticorpos anti-LVPR em amostras de plasma seminal, possivelmente em função da compartimentalização do vírus. Intermitência viral foi observada em ambas as amostras biológicas, independentemente do estágio de infecção. Conclui-se que a resposta imunológica em reprodutores com infecção recente LVPR é mais acentuada do que em animais cronicamente infectados. E, independentemente do estágio da infecção há uma flutuação nos níveis de anticorpos, sendo, portanto, um fator de risco, em virtude da existência de amostras falso-negativo ao realizar o diagnóstico.

Natural association between bovine leukemia virus and reproductive infectious diseases

Background: Enzootic bovine leukosis (EBL) is a widespread infectious disease caused by the bovine leukemia virus (BLV), which results in immune system dysfunction. The resulting immunosuppression may lead to an increased prevalence of other diseases. Dairy cows infected have altered immune function associated with decreased milk production and shortened lifespan and decreased immune response to immunization. BLV infection, however, is often asymptomatic, so any connection between subclinical infection and common reproductive diseases remains unknown. This study aimed to describe the relationship between naturally occurring subclinical BLV and infectious reproductive diseases seroconversion in the field. Materials, Methods & Results: The diseases investigated included Bovine viral diarrhoea (BVD), Bovine alfaherpesvirus 1 (BoHV-1), Bovine gammaherpesvirus 4 (BoHV-4), Chlamydiosis, Leptospirosis, Brucellosis and Neosporosis in dairy cattle. Six hundred fifty-five sera samples from the northern and south-central regions of Uruguay, from asymptomatic female Holstein and Holstein crosses without a history of vaccination against reproductive diseases were processed using reference diagnostic methods (Seronautalization, ELISA, MAT, Rose Bengal Plate test). The seroprevalence of BLV was 20.0%. Seroprevalence of reproductive diseases BVD, BoHV-1 and BoHV-4 were 99.3%, 41.2% and 27.3% of the populations, respectively, and the total seroprevalence of Leptospirosis, bovine Neospora caninum and Chlamydiosis were 19.8%, 29.8% and 33.0% respectively. The results revealed positive associations between naturally contracted BLV and the presence of antibodies against BoHV-1 (P = 0.002), as well as between naturally contracted BLV and presence of antibodies against Leptospira spp. (P = 0.028). Discussion: BLV infection can impact innate and adaptive immune system cells and alter the proper functioning of uninfected cells. BLV infection may also induce changes in the complex balance of cytokine expression, cell proliferation, and programmed cell death in T- and B-lymphocytes, which is critical for immune competence and effective response to infectious challenges. The progression of BLV infection has a substantial effect on host defense mechanisms. Indeed, lowmagnitude serologic responses to a commercial foot-and-mouth disease vaccine and a J5 Escherichia coli vaccine have been observed. These results are supported by recent trial studies showing a reduced immune response to vaccination against BoHV-1 and Leptospira spp. in asymptomatic animals infected with BLV. These are 2 of the most prevalent infectious reproductive diseases in cattle worldwide, and our results provide evidence that a link between BLV and susceptibility to these diseases may exist. Although there is evidence of the co-occurrence of these diseases, it remains unknown whether there is a direct or indirect effect of BLV on infertility, embryonic loss, or abortion. Another possibility is that natural infection with these reproductive pathogens (BoHV-1, Leptospira, or others) promotes BLV expression, negatively affecting the farms where these pathogens are endemic. Considering the high seroprevalence of BLV in dairy herds in North and South America where the infection is endemic, it was explored BLV's role as an immunosuppressant by quantifying its co-occurrence with diseases that affect reproductive performance in breeding herds. Future work should clarify the role of BLV and the co-occurring pathogens in causing infertility or abortions.

Neuropathology of natural canid alphaherpesvirus 1 infection in dogs

The neuropathology of canid alphaherpesvirus 1 (CaHV-1) infection in dogs has been reported, but the importance of the cerebellar lesions in the routine diagnosis remains relatively understated in the veterinary literature. Here we characterize the neuropathology of natural CaHV-1 infection in 25 dogs. The disease affected predominantly male dogs (22 cases). Clinical signs in 15 cases varied and sudden death was reported in 10 cases. All dogs had the typical areas of necrosis and hemorrhage in multiple organs with eosinophilic intranuclear viral inclusions in epithelial and/or inflammatory cells in multiple organs in 21 cases. Cerebral swelling and reddening were reported in 3 cases. Neurohistologic changes consisted of mild lymphoplasmacytic meningoencephalitis with occasional vasculitis and widespread glial nodules. Rare intranuclear viral inclusions (4 cases) were observed in endothelial cells or inflammatory cells in glial nodules. Cerebellum was examined in 11 cases and had extensive segmental necrosis of Purkinje cell layer (8 cases) and granule neurons in the internal (9 cases) or external (2 cases) granular cell layer. Intranuclear viral inclusions were observed in 8 cases and occurred in necrotic granule neurons in the inner granular cell layer (7 cases), outer granular cell layer (2 cases), Purkinje neurons (1 case), or endothelial cells (1 case). Diagnostic confirmation in 23 cases was based on routine histology and fluorescent antibody testing for CaHV-1 on fresh or frozen tissue samples. A routine diagnosis was based solely on the visualization of intranuclear viral inclusions in 2 cases.(AU)

Bioactivity of substances isolated from natural products on mollusks Biomphalaria glabrata (Say, 1818) (Planorbidae): a review / Bioatividade de substâncias isoladas de produtos naturais em moluscos Biomphalaria glabrata (Say, 1818) (Planorbidae): uma revisão

Schistosomiasis is a neglected tropical disease caused by parasitic worms of several species of the genus Schistosoma. Transmission occurs by parasitic larvae that stay in freshwater snails of the genus Biomphalaria. Thus, the search for new products that are biodegradable has increased the interest in products of plant origin. The aim of this article is to review the isolated substances from natural products that showed molluscicidal activity against the species Biomphalaria glabrata in order to reevaluate the most promising prototypes and update the progress of research to obtain a new molluscicide. We perform searches using scientific databases, such as Scientific Electronic Library Online (SciELO), Google schoolar, PUBMED, Web of Science and Latin American and Caribbean Literature on Health Sciences (LILACS). From 2000 to 2022, using the keywords "isolated substances", "molluscicidal activity" and "Biomphalaria glabrata". In the present study, it was possible to observe 19 promising molluscicidal molecules with a lethal concentration below 20 µg/mL. Of these promising isolates, only 5 isolates had the CL90 calculated and within the value recommended by WHO: Benzoic acid, 2',4',6'-Trihydroxydihydrochalcone, Divaricatic acid, Piplartine and 2-hydroxy-1,4-naphthoquinone (Lapachol). We conclude that beyond a few results in the area, the researches don't follow the methodological pattern (exposure time and measure units, toxicity test), in this way, as they don't follow a pattern on the result's exposure (LC), not following, in sum, the recommended by WHO.

A esquistossomose é uma doença tropical negligenciada causada por vermes parasitas de várias espécies do gênero Schistosoma. A transmissão ocorre por larvas parasitas que ficam em caramujos de água doce do gênero Biomphalaria. Assim, a busca por novos produtos biodegradáveis ​​tem aumentado o interesse por produtos de origem vegetal. O objetivo deste artigo é revisar as substâncias isoladas de produtos naturais que apresentaram atividade moluscicida contra a espécie Biomphalaria glabrata a fim de reavaliar os protótipos mais promissores e atualizar o progresso das pesquisas para a obtenção de um novo moluscicida. Realizamos buscas em bases de dados científicas, como Scientific Electronic Library Online (SciELO), Google acadêmico, PUBMED, Web of Science e Literatura Latina-Americana e do Caribe em Ciências da saúde (LILACS). De 2000 a 2022, utilizando as palavras-chave "substâncias isoladas", "atividade moluscicida" e "Biomphalaria glabrata". No presente estudo, foi possível observar 19 moléculas moluscicidas promissoras com concentração letal abaixo de 20 µg/mL. Destes isolados promissores, apenas 5 isolados tiveram o CL90 calculado e dentro do valor recomendado pela OMS: ácido benzóico, 2',4',6'-triidroxidihidrocalcona, ácido divaricático, piplartina e 2-hidroxi-1,4-naftoquinona (Lapachol). Concluímos que além de poucos resultados na área, as pesquisas não seguem o padrão metodológico (tempo de exposição e unidades de medida, teste de toxicidade), assim como não seguem um padrão na exposição do resultado (CL), não seguindo, em suma, o recomendado pela OMS.

Agro-industrial residues in the control of gastrointestinal nematodes of small ruminants and fertilization of forages / Resíduos agroindustriais no controle de nematóides gastrointestinais de pequenos ruminantes e adubação de forrageiras

Verminosis in small ruminants can render rural activity impractical, which is still controlled through the administration of anthelmintics. The present study evaluated four agro-industrial residues as fertilizer in the control of GIN of small ruminants in the free-living stage. Crab shell, manipueira, biochar, and organomineral residues were used in pots (5.0 kg of soil) and cultivated with Massai grass (Megathyrsus maximum cv. Massai). Further, the pots were contaminated with feces from sheep carrying a natural multispecific infection by GIN. Next, the residues were applied individually to the soil, with 50 mL/pot for liquid and 50 g/pot for solid residues. Treatment with manipueira showed the lowest number of L3.kg MS-1 recovered from the grass (202.44), when compared with treatments using organomineral (823.89) and biochar (689.34). However, there was no statistically significant difference (P < 0.05) between the four treatments as compared to the control group. These agro-industrial residues can be used as organic fertilizers; however, these will not help in the control of GIN in sheep.

A verminose em pequenos ruminantes é uma doença que pode inviabilizar a atividade rural. Embora o sucesso no controle dos nematóides gastrointestinais de pequenos ruminantes dependa da adoção associada de diferentes estratégias e tecnologias, ainda é possível notar a escolha, por parte de alguns ovinocaprinocultores, pelo tratamento exclusivamente baseado na administração de anti-helmínticos aos animais. Contudo, é no ambiente que encontramos um elevado número de formas imaturas de nematóides gastrointestinais. Objetivou-se com o presente trabalho avaliar quatro resíduos agroindustriais como adubo no controle dos nematóides gastrointestinais de pequenos ruminantes, em estádio de vida livre. Casca de caranguejo, manipueira, biochar e organomineral foram os resíduos empregados nos vasos com capacidade média de 5,0 kg de solo, onde foi cultivado o capim Massai (Megathyrsus maximus cv. Massai). Após corte de uniformização do capim, os vasos foram contaminados utilizando-se fezes de ovinos portadores de infecção natural multiespecífica por nematóides gastrointestinais. E, na sequência, os resíduos foram aplicados individualmente no solo, na quantidade de 50 mL/vaso para o resíduo líquido e 50 g/vaso para os sólidos. O tratamento com a manipueira foi o que apresentou o menor número de larvas infectantes de terceiro estádio (L3) recuperado do capim (202,44 L3.kg MS-1) quando comparado com os tratamentos utilizando organomineral (823,89 L3.kg MS-1) e biochar (689,34 L3.kg MS-1). Contudo, não houve diferença estatística significativa (P < 0,05) entre os quatro tratamentos testados quando comparado ao grupo controle. Os resíduos agroindustriais podem ser utilizados como adubos orgânicos, mas não vão auxiliar no controle dos nematóides gastrintestinais dos ovinos.

Essential oils isolated from popular medicinal plants and spices as alternative antimicrobial and antibiofilm compounds against the pig pathogen Actinobacilluspleuropneumoniae / Óleos essenciais isolados de plantas medicinais e temperos populares como compostos antimicrobianos e antibiofilme alternativos contra o patógeno de suínosActinobacillus pleuropneumoniae

ABSTRACT: Actinobacilluspleuropneumoniae is the causative agent of swine pleuropneumonia, a contagious respiratory disease associated with high morbidity and economic losses. While antibiotic therapy helps to control the spreading of the pathogen on the farm, resistance to several classes of antibiotics were reported, and treatment can be impaired by the bacterial ability to form biofilms. This increases the need for alternative therapy approaches, including the use of natural compounds with antimicrobial and/or antibiofilm activities. In this research we analyzed, by the broth microdilution method, the inhibitory and bactericidal activities of the essential oils obtained from eighteen Brazilian popular medicinal plants or spices against clinical isolates of Actinobacilluspleuropneumoniae. After that, sub-inhibitory concentrations of active oils were tested for their antibiofilm effects, analyzed by the crystal violet method. Among the eighteen oils tested, eight (extracted from cinnamon, coriander, peppermint, spearmint, thyme, marjoram, eucalyptus, and laurel) presented bacteriostatic and bactericidal activities against all isolates, and subinhibitory concentrations of five of them disrupted up to 80% preformed biofilms, and significantly inhibited biofilm formation. The chemical composition of such oils was assessed by gas chromatography/mass spectrometry (GC/MS) and is presented, indicating that their bactericidal and antibiofilm properties were mostly associated with the presence of monoterpenes and phenylpropanoids. To our knowledge, this is the first report of essential oils with potential to control environmental contamination and animal infection with A. pleuropneumoniae, representing an alternative to increasing levels of antibiotic resistance.

RESUMO: Actinobacillus pleuropneumoniae é o agente causador da pleuropneumonia suína, uma doença respiratória contagiosa associada à alta morbidade e perdas econômicas. Embora a antibioticoterapia ajude a controlar a disseminação do patógeno na fazenda, a resistência a várias classes de antibióticos tem sido relatada e o tratamento pode ser prejudicado pela capacidade bacteriana de formar biofilmes. Isso aumenta a necessidade de abordagens terapêuticas alternativas, que incluem o uso de compostos naturais com atividades antimicrobianas e/ou antibiofilme. Neste trabalho, analisamos, pelo método de microdiluição em caldo nutriente, os efeitos inibitórios e bactericidas dos óleos essenciais obtidos de dezoito plantas medicinais brasileiras populares e/ou temperos contra isolados clínicos de Actinobacillus pleuropneumoniae. Então, concentrações subinibitórias dos óleos ativos foram testados quanto a suas atividades antibiofilme, analisados pelo método do cristal violeta. Dentre os dezoito óleos testados, oito (extraídos da canela, coentro, hortelã-pimenta, hortelã, tomilho, manjerona, eucalipto e louro) apresentaram atividade bacteriostática e bactericida contra todos os isolados, e concentrações subinibitórias de cinco deles romperam biofilmes pré-formados em até 80%, além de inibirem fortemente a formação de biofilmes. A composição química desses óleos foi avaliada por cromatografia gasosa/espectrometria de massa (CG/MS) e é apresentada, indicando que suas propriedades bactericidas e antibiofilme estavam principalmente associadas à presença de monoterpenos e fenilpropanóides. Este é o primeiro relato de óleos essenciais com potencial para controlar a contaminação ambiental e infecção animal por A. pleuropneumoniae, representando uma alternativa contra o crescente aumento de resistência bacteriana aos antibióticos.

A threefold approach including quantum chemical, molecular docking and molecular dynamic studies to explore the natural compounds from Centaurea jacea as the potential inhibitors for COVID-19 / Uma abordagem tríplice incluindo química quântica, docking molecular e estudos dinâmicos moleculares para explorar os compostos naturais de Centaurea jacea como inibidores potenciais para Covid-19

Abstract In the current report, we studied the possible inhibitors of COVID-19 from bioactive constituents of Centaurea jacea using a threefold approach consisting of quantum chemical, molecular docking and molecular dynamic techniques. Centaurea jacea is a perennial herb often used in folk medicines of dermatological complaints and fever. Moreover, anticancer, antioxidant, antibacterial and antiviral properties of its bioactive compounds are also reported. The Mpro (Main proteases) was docked with different compounds of Centaurea jacea through molecular docking. All the studied compounds including apigenin, axillarin, Centaureidin, Cirsiliol, Eupatorin and Isokaempferide, show suitable binding affinities to the binding site of SARS-CoV-2 main protease with their binding energies -6.7 kcal/mol, -7.4 kcal/mol, -7.0 kcal/mol, -5.8 kcal/mol, -6.2 kcal/mol and -6.8 kcal/mol, respectively. Among all studied compounds, axillarin was found to have maximum inhibitor efficiency followed by Centaureidin, Isokaempferide, Apigenin, Eupatorin and Cirsiliol. Our results suggested that axillarin binds with the most crucial catalytic residues CYS145 and HIS41 of the Mpro, moreover axillarin shows 5 hydrogen bond interactions and 5 hydrophobic interactions with various residues of Mpro. Furthermore, the molecular dynamic calculations over 60 ns (6×106 femtosecond) time scale also shown significant insights into the binding effects of axillarin with Mpro of SARS-CoV-2 by imitating protein like aqueous environment. From molecular dynamic calculations, the RMSD and RMSF computations indicate the stability and dynamics of the best docked complex in aqueous environment. The ADME properties and toxicity prediction analysis of axillarin also recommended it as safe drug candidate. Further, in vivo and in vitro investigations are essential to ensure the anti SARS-CoV-2 activity of all bioactive compounds particularly axillarin to encourage preventive use of Centaurea jacea against COVID-19 infections.

Resumo No presente relatório, estudamos os possíveis inibidores de Covid-19 de constituintes bioativos de Centaurea jacea usando uma abordagem tripla que consiste em técnicas de química quântica, docking molecular e dinâmica molecular. Centaurea jacea é uma erva perene frequentemente usada em remédios populares de doenças dermatológicas e febre. Além disso, as propriedades anticâncer, antioxidante, antibacteriana e antiviral de seus compostos bioativos também são relatadas. A Mpro (proteases principais) foi acoplada a diferentes compostos de Centaurea jacea por meio de docking molecular. Todos os compostos estudados, incluindo apigenina, axilarina, Centaureidina, Cirsiliol, Eupatorina e Isokaempferide, mostram afinidades de ligação adequadas ao sítio de ligação da protease principal SARS-CoV-2 com suas energias de ligação -6,7 kcal / mol, -7,4 kcal / mol, - 7,0 kcal / mol, -5,8 kcal / mol, -6,2 kcal / mol e -6,8 kcal / mol, respectivamente. Dentre todos os compostos estudados, a axilarina apresentou eficiência máxima de inibidor, seguida pela Centaureidina, Isokaempferida, Apigenina, Eupatorina e Cirsiliol. Nossos resultados sugeriram que a axilarina se liga aos resíduos catalíticos mais cruciais CYS145 e HIS41 do Mpro, além disso a axilarina mostra 5 interações de ligações de hidrogênio e 5 interações hidrofóbicas com vários resíduos de Mpro. Além disso, os cálculos de dinâmica molecular em uma escala de tempo de 60 ns (6 × 106 femtossegundos) também mostraram percepções significativas sobre os efeitos de ligação da axilarina com Mpro de SARS-CoV-2 por imitação de proteínas como o ambiente aquoso. A partir de cálculos de dinâmica molecular, os cálculos RMSD e RMSF indicam a estabilidade e dinâmica do melhor complexo ancorado em ambiente aquoso. As propriedades ADME e a análise de previsão de toxicidade da axilarina também a recomendaram como um candidato a medicamento seguro. Além disso, as investigações in vivo e in vitro são essenciais para garantir a atividade anti-SARS-CoV-2 de todos os compostos bioativos, particularmente a axilarina, para encorajar o uso preventivo de Centaurea jacea contra infecções por Covid-19.

Chronic superficial infection in a dog caused by multidrug-resistant Pseudomonas aeruginosa

Background: Pseudomonas aeruginosa is a gram-negative aerobic bacterium and non-glucose fermenting, that usually causes opportunistic infections in animals, including humans. It is rarely involved in primary disease. The antibioticresistant bacterial strains are mainly developed due to the inappropriate use of antibiotics, however treating P. aeruginosa infections can be difficult owing to their natural resistance to antibiotics. Furthermorer resistant microorganisms such as P. aeruginosa grow by developing biofilms. Inaccurate diagnoses and absence of adequate microbiological tests can cause difficulties in resolving cases. This report describes a case of chronic superficial infection in a bitch caused by multidrugresistant Pseudomonas aeruginosa (MDR-PA). Case: A 6-year-old bitch Shih Tzu, initially presented with an exudative erythematous lesion in the snout region, which progressed to deep lesions, and spread to the back and limbs; furthermore, the animal always experienced a fever before new wounds emerged. Lesion samples, collected using a swab and processed at the Veterinary Microbiology Laboratory of the Federal University of Jatai (UFJ), revealed the presence of Pseudomonas aeruginosa. The isolate was multidrug-resistant and a carrier of TEM and ppyR genes. In the diffusion disk antibiogram, the isolate was found resistant to 14 different antibiotics belonging to 6 classes. Antimicrobial resistance was also tested using the minimum inhibitory concentration (MIC) test against imipenem, ceftazidime, ciprofloxacin, ticarcillin + clavulanic acid and aztreonam present in the MIC test strip. Treatment with amikacin and muporicin proved to be effective; however, owing to lesions extending to the face and palpebral involvement, the animal lost its eyeballs. Discussion: Pseudomonas aeruginosa is frequently associated with nosocomial infections mainly affecting immunosuppressed patients. Among the antibiotics tested, the group with the highest number of ineffective antibiotics was beta-lactams, where sensitivity was only observed for ticarcillin and ceftazidime. Recent studies have demonstrated that ceftazidime can reduce biofilm volume, inhibit motility, and repress the expression of genes associated with bacterial adhesion in P. aeruginosa. Therefore, the production of biofilm in P. aeruginosa is an important virulence factor as it facilitates a stable environment for the microorganism, which protects the bacteria from contact with antimicrobials. In addition, prolonged exposure to a wide variety of antimicrobials creates an environment of selective pressure between microorganisms, facilitating the emergence of multidrug-resistant strains. Furthermore, it is now well recognized that low doses of antibiotics, administered during continuous and fluctuating treatments, can stimulate biofilm establishment and are partly responsible for biofilm-specific antimicrobial tolerance. The resistance profile of P. aeruginosa isolated from dogs varies considerably, and the presence of isolates with a possible biofilm production capacity represents a challenge for the interpretation of the antimicrobial susceptibility profile. Culture and antibiogram is fundamentally important, both clinically and in environmental monitoring, in addition to the use of antibiogram data for decision making in clinical treatment.

Neoctangium travassosi (Digenea: Microscaphidiidae) in sea turtles from South America / Neoctangium travassosi (Digenea: Microscaphidiidae) em tartarugas marinhas da América do Sul

Sea turtles are endangered animals that present cosmopolitan distribution. Anthropic actions have been considered important causes for the reduction of sea turtle population, but natural aspects such as parasitism may also contribute to their decline. This study aimed to report the occurrence of parasites in stranded dead sea turtles found in an area known as Potiguar Basin, northeastern Brazil, from 2010 to 2019. They were identified and classified according to the carapace length. At post-mortem analyses all organs were examined, parasites collected and morphologically identified. Ecological parasitic indexes as prevalence (P), mean intensity (MI) and mean abundance (MA) were calculated. A total of 80 Chelonia mydas and 5 Eretmochelys imbricata were assessed. Neoctangium travassosi was detected in both species presenting P = 20%, MI = 4.19 and MA = 0.84 for C. mydas and P = 60%, MI = 1.67 and MA = 1.0 for E. imbricata. This is the first report of N. travassosi parasitizing E. imbricata in South America. Finally, the retrieval of these parasites is a warning regarding the need for further studies to assess the impact of this parasitism on the health and conservation of sea turtles.

As tartarugas marinhas são animais ameaçados de extinção que apresentam distribuição cosmopolita. Ações antrópicas têm sido consideradas causas importantes para a redução da população de tartarugas marinhas, mas aspectos naturais, como o parasitismo, também, podem contribuir para o seu declínio. Este estudo objetivou relatar a ocorrência de parasitos em tartarugas marinhas, encalhadas mortas, encontradas em uma área conhecida como Bacia Potiguar, nordeste do Brasil, de 2010 a 2019. Essas foram identificadas e classificadas de acordo com o comprimento da carapaça. Nas análises post mortem, todos os órgãos foram examinados, e os parasitos coletados e identificados morfologicamente. Foram calculados índices parasitários ecológicos, como prevalência (P), intensidade média (IM) e abundância média (AM). Um total de 80 Chelonia mydas e cinco Eretmochelys imbricata foi estudado. Neoctangium travassosi foi detectado em ambas as espécies, apresentando P = 20%, MI = 4,19 e MA = 0,84 para C. mydas e P = 60%, IM = 1,67 e AM = 1,0 para E. imbricata. Este é o primeiro relato de N. travassossi parasitando E. imbricata na América do Sul. Conclui-se que a recuperação desses parasitos soa como um alerta para a necessidade de mais estudos para avaliar o impacto desse parasitismo na saúde e na conservação das tartarugas marinhas.

First record of Neoechinorhynchus buttnerae and Piscinoodinium pillulare infection in Colossoma macropomum in the state of Tocantins, Brazil / Primeiro registro de infecção por Neoechinorhynchus buttnerae e Piscinoodinium pillulare em Colossoma macropomum no estado do Tocantins, Brasil

This study aims to report the occurrence of two important parasites in farmed tambaqui Colossoma macropomum in the state of Tocantins, the acanthocephalan Neoechinorhynchus buttnerae and the dinoflagellate protozoan Piscinoodinium pillulare, also suggesting the main treatments to control them. The fish sampled for the study were infected by N. buttnerae, and P. pillulare, with prevalence from 100% and mean intensity from 51.4 to 354,264, respectively. This was the first report on the occurrence of such parasites in C. macropomum in the state of Tocantins. We emphasize the need to adopt good farm management and biosecurity practices to prevent pathogenic agents to enter or leave a property. Reported treatments with synthetic and natural products with positive results are also suggested to treat against those parasites in farmed C. macropomum.(AU)

Este estudo teve como objetivo relatar a ocorrência de dois importantes parasitos em tambaqui Colossoma macropomum cultivado no estado de Tocantins, o acantocéfalo Neoechinorhynchus buttnerae e o protozoário dinoflagelado Piscinoodinium pillulare e também destacar os principais tratamentos para controlá-los. Os peixes examinados estavam infectados por N. buttnerae e P. pillulare, com prevalência de 100% e intensidade média de 51,4 a 354.264,4, respectivamente. Este foi o primeiro relato da ocorrência desses parasitos para C. macropomum no estado de Tocantins. Destaca-se a necessidade da adoção de boas práticas de manejo na produção e de manejo sanitário, para evitar a contaminação da propriedade e a transmissão para pisciculturas vizinhas. Em relação ao controle dessas parasitoses são apresentados os tratamentos com resultados positivos para C. macropomum, com o uso de produtos sintéticos e naturais.(AU)

Comparative Study on the Predominance of Lactobacillus spp. and Escherichia Coli in Healthy vs Colibacillosis Diseased Broilers

This study aims to identify relative proportions of beneficial and pathogenic bacteria in the gut of broilers and risk factors that may be contributing to the development of colibacillosis disease in broiler farms of District Kasur, Punjab, Pakistan. For this, 10 healthy and 10 colibacillosis affected broiler farms were surveyed for ileum and blood sample collection along with data regarding farm management, antibiotic use and hygiene practices. Lactobacillus and Escherichia coli number was estimated using Miles and Misra method and colibacillosis was confirmed by Congo red dye assay. Lactobacillus and E. coli were identified biochemically. For risk factors analysis chi-square analysis was performed to find any significant association between the health status of the farm and risk factors. Results showed during disease and healthy conditions Lactobacillus and Escherichia coli counts differ significantly (p<0.05). E. coli counts (106-108 to 107-109) increased (p<0.05) about three folds and Lactobacillus counts decrease (106-108 to 105-107) about four folds in disease conditions. Risk factor analysis showed colibacillosis disease was significantly associated (p<0.05) with non-vaccinated flocks, natural ventilation systems, rodent presence and the lack of outfit disinfection or change by workers when moving between different houses. It is concluded that E. coli and Lactobacillus work antagonistically to each other. However, further research is necessary to determine the exact mechanisms by which E. coli and Lactobacillus influence the development of colibacillosis. While Lactobacillus as probiotic may help with prevention, good hygiene and management practices are still crucial in preventing the spread of disease.(AU)

Aspergilose em calopsita (Nymphicus hollandicus) / Aspergillosis in a cockatiel (Nymphicus hollandicus)

Background: Aspergillosis is a defiance in avian medicine, however between of Aspergillus species, 3 are the most frequently observed (A. flavus, A. fumigatus and A. niger). Psittaciformes present medical conditions related to the 3 species, however A. fumigatus is the most observed. The defiance of the diagnosis is related to fungus characteristics (they are saprophytic in healthy birds), the clinical signs and lesions, which are common to various respiratory diseases, and the difficulty to access specific diagnostic methods. Thus, we report a case of aspergillosis in a cockatiel, whose clinical diagnosis was confirmed by fungal culture of cloacal swab, and treatment with a fungistatic drug. Case: A female cockatiel, 2-month-old, not dewormed, presented diarrhea, sneezing and difficulty to gain weight, although eating well (extruded feed and supplementation with fruits and vegetables). At a clinical examination revealed body score (BCS) of 3 (scale 1 at 5), cloacal temperature (CT) of 40 ºC, dilated nostrils and edema in the left eye region associated to evident hyperemia. Due to it has lived with another animal, was inferred that was hypovitaminosis A, by food dispute and/ or dominance, or endoparasitosis, being prescribed vitamin A supplementation and deworming with ivermectin. The bird returned 15 days later, showing maintenance of weight loss, BCS 2, ruffled feathers and areas suggestive of apteriolysis in both forelimbs, respiratory difficulty and pulmonary and cranial air sacs rales, in addition presented wound in the pygostyle region and ventus, occasioned by attempted attack of a dog, and aggravated by self-mutilation. Due the worsening of the clinical condition, an oral cavity and cloacal swabs were collected to bacteriological and fungal culture. As treatment was prescribed: a natural tranquilizer, seeking to control the self-mutilation; doxycycline oral suspension, to treat a probable bacterial respiratory infection; and oral itraconazole suspension, manipulated and with a dose calculated by allometric scaling, being placed an elizabethan collar in order to prevent biting the pygostyle and ventus. With a positive result of the cloacal swab fungal culture for A. fumigatus, the doxycycline was discontinued. After the first 30 days of treatment with itraconazole, the patient returns to alert, weighing 95 g, BCS 3, with absence of respiratory signs and with signs of recent molting process. New swabs (oral and cloacal) were collected for repetition of bacterial and fungal cultures, being maintained the treatment with itraconazole. The results of both bacterial and fungal cultures were negative. At the end of the treatment with itraconazole, the patient returns demonstrating weight gain, EEC 3, CT of 42ºC, uniform feather coverage and absence of any respiratory sign, being released by the clinician. Discussion: A preponderant factor in the treatment of various respiratory diseases in birds is the diagnosis, since many of diseases present the same clinical signs. Thus, the use of fungal culture was essential to determine the origin of the signs. Still, even with an accurate diagnosis, the clinician often comes up against the difficulty of treatment, especially in cases of mycotic infections. The knowledge about the best drug, considering the pharmacology, as well as the availability of concentrations and dosages marketed, in addition to the long period of treatment, can difficult or make impossible the case resolution. In fact, this is confirmed by the literature, where the most of aspergillosis diagnosis are post mortem, resulting from the delay in seeking veterinary care or in the administration of ineffective drugs. Thus, with this report, we sought to present a diagnostic tool, sometimes neglected, and a compilation with the main drugs and their effectiveness in the treatment of aspergillosis in Psittaciformes.

First molecular detection of Leishmania (Leishmania) infantum chagasi in a domestic cat (Felis catus) from an urban area in eastern Amazon

Background: Domestic cats have been implicated as accidental hosts of Leishmania sp. However, in recent years, the recurrent description of new cases in endemic and nonendemic areas draw attention to the potential epidemiological role of cats as reservoir hosts. Although dogs are considered urban reservoirs, cats could act as a secondary natural reservoirs in these areas. Thus, feline leishmaniasis has become an emerging disease in several countries worldwide. Case presentation: This study aimed to describe the first case of feline leishmaniasis in a stray animal that presented lesions compatible with the disease in Belém, Pará, Brazil, an important urban area in eastern Amazon. Serological tests for Leishmania infantum (ELISA and IFA) were nonreactive, whereas histopathological examination indicated infectious dermatitis caused by Leishmania spp. or Toxoplasma gondii. Cytopathological study of lesion aspirate confirmed the presence of Leishmania sp. amastigotes within macrophages. Finally, molecular analyses revealed that the feline infection was caused by Leishmania (Leishmania) infantum chagasi. Conclusion: To the best of the authors' knowledge, this study reports the first case of natural infection by Leishmania (Leishmania) infantum chagasi in a feline from eastern Amazon. These findings suggest domestic cats as potential secondary reservoir hosts of Leishmania spp. in Belém, which reinforces the importance of further epidemiological investigation of feline leishmaniasis, especially in urban areas with human cases.(AU)

Genetic diversity of Anaplasma marginale in calves under natural transmission conditions in the Northeast region of Pará / Diversidade genética de Anaplasma marginale em bezerros sob condições de transmissão natural na região Nordeste do Pará

The objective of the present study was to detect the genetic diversity of Anaplasma marginale strains in naturally infected calves from a rural property located in the northeastern region of the state of Pará, Eastern Amazon, which has a history of mortality due to anaplasmosis. Fourteen calves positive for A. marginale were selected using a semi-nested polymerase chain reaction for the target msp1α gene, with asymptomatic (n=3) and symptomatic (n=11) infections. After sequencing the samples, two genotypes were verified in the E and C regions and the structures in tandem repeats were determined. Nine different strains were found: eight related to the E genotype (α-ß-ß-Γ = one animal, asymptomatic; 16-F-17-F-F = two animals, symptomatic; α-ß-F-F-F-F = one animal, asymptomatic; 31-62-62-61 = one animal, symptomatic; τ-10-3 = three animals, two symptomatic and one asymptomatic; α-ß-ß-ß = one animal, symptomatic; τ-22 -13-18 = two animals, both symptomatic; ß-ß-ß-BRA1-31 = two animals, both symptomatic), and one related to genotype C (23-24-25-31-27-27 = one animal, asymptomatic). Genotype E was predominant in 92.86% of the samples (13/14), followed by genotype C (7.14%). This study made it possible to detect the genetic diversity of A. marginale in calves from the selected dairy farm, in addition to identifying the BRA1 sequence in the animals of the present study, which was recently diagnosed in Minas Gerais, demonstrating the dispersion of A. marginale strains in herds from different Brazilian states. Genetic diversity of A. marginale was observed in both symptomatic and asymptomatic calves. There were no significant differences when clinical signs were compared to the genotype verified in the infected animals. The prevalence of pathogenicity was not observed.

O objetivo do presente trabalho foi detectar a diversidade genética de cepas de Anaplasma marginale em bezerros naturalmente infectados oriundos de uma propriedade rural localizada na região nordeste do estado do Pará, Amazônia Oriental, a qual apresentava histórico de mortalidade devido à anaplasmose. Foram selecionados 14 bezerros positivos para A. marginale pela técnica de semi-nested PCR (nPCR) para o alvo no gene msp1α, com infecção assintomática (n=3) e sintomáticos (n=11). Após o sequenciamento das amostras foram verificados dois genótipos nas regiões E e C, e determinadas as estruturas em tandem repeats. Nove diferentes estirpes foram encontradas, sendo oito relacionadas ao genótipo E (α-ß-ß-Γ = um animal, assintomático; 16-F-17-F-F = dois animais, sintomáticos; α-ß-F-F-F-F = um animal, assintomático; 31-62-62-61 = um animal, sintomático; τ-10-3 = três animais, dois sintomáticos e um assintomático; α-ß-ß-ß = um animal, sintomático; τ-22-13-18 = dois animais, sintomáticos; ß-ß-ß-BRA1-31 = dois animais, sintomáticos) e uma relacionada ao genótipo C (23-24-25-31-27-27 = um animal, assintomático). O genótipo E foi predominante em 92,86% das amostras (13/14), seguido pelo genótipo C (7,14%). O estudo possibilitou a detecção da diversidade genética de A. marginale em bezerros dessa propriedade leiteira, além de identificar a sequência BRA1 nos animais do presente estudo, a qual foi diagnosticada recentemente em Minas Gerais, o que demonstra a dispersão das estirpes de A. marginale nos rebanhos de diferentes estados brasileiros. A diversidade genética de A. marginale foi observada tanto em bezerros sintomáticos quanto em assintomáticos e não houve diferença significativa quando se comparou os sinais clínicos ao genótipo verificado no animal infectado, não observando a prevalência de patogenicidade de estirpes.

Essential oils isolated from popular medicinal plants and spices as alternative antimicrobial and antibiofilm compounds against the pig pathogen Actinobacilluspleuropneumoniae / Óleos essenciais isolados de plantas medicinais e temperos populares como compostos antimicrobianos e antibiofilme alternativos contra o patógeno de suínosActinobacillus pleuropneumoniae

Actinobacilluspleuropneumoniae is the causative agent of swine pleuropneumonia, a contagious respiratory disease associated with high morbidity and economic losses. While antibiotic therapy helps to control the spreading of the pathogen on the farm, resistance to several classes of antibiotics were reported, and treatment can be impaired by the bacterial ability to form biofilms. This increases the need for alternative therapy approaches, including the use of natural compounds with antimicrobial and/or antibiofilm activities. In this research we analyzed, by the broth microdilution method, the inhibitory and bactericidal activities of the essential oils obtained from eighteen Brazilian popular medicinal plants or spices against clinical isolates of Actinobacilluspleuropneumoniae. After that, sub-inhibitory concentrations of active oils were tested for their antibiofilm effects, analyzed by the crystal violet method. Among the eighteen oils tested, eight (extracted from cinnamon, coriander, peppermint, spearmint, thyme, marjoram, eucalyptus, and laurel) presented bacteriostatic and bactericidal activities against all isolates, and subinhibitory concentrations of five of them disrupted up to 80% preformed biofilms, and significantly inhibited biofilm formation. The chemical composition of such oils was assessed by gas chromatography/mass spectrometry (GC/MS) and is presented, indicating that their bactericidal and antibiofilm properties were mostly associated with the presence of monoterpenes and phenylpropanoids. To our knowledge, this is the first report of essential oils with potential to control environmental contamination and animal infection with A. pleuropneumoniae, representing an alternative to increasing levels of antibiotic resistance.

Actinobacillus pleuropneumoniae é o agente causador da pleuropneumonia suína, uma doença respiratória contagiosa associada à alta morbidade e perdas econômicas. Embora a antibioticoterapia ajude a controlar a disseminação do patógeno na fazenda, a resistência a várias classes de antibióticos tem sido relatada e o tratamento pode ser prejudicado pela capacidade bacteriana de formar biofilmes. Isso aumenta a necessidade de abordagens terapêuticas alternativas, que incluem o uso de compostos naturais com atividades antimicrobianas e/ou antibiofilme. Neste trabalho, analisamos, pelo método de microdiluição em caldo nutriente, os efeitos inibitórios e bactericidas dos óleos essenciais obtidos de dezoito plantas medicinais brasileiras populares e/ou temperos contra isolados clínicos de Actinobacillus pleuropneumoniae. Então, concentrações subinibitórias dos óleos ativos foram testados quanto a suas atividades antibiofilme, analisados pelo método do cristal violeta. Dentre os dezoito óleos testados, oito (extraídos da canela, coentro, hortelã-pimenta, hortelã, tomilho, manjerona, eucalipto e louro) apresentaram atividade bacteriostática e bactericida contra todos os isolados, e concentrações subinibitórias de cinco deles romperam biofilmes pré-formados em até 80%, além de inibirem fortemente a formação de biofilmes. A composição química desses óleos foi avaliada por cromatografia gasosa/espectrometria de massa (CG/MS) e é apresentada, indicando que suas propriedades bactericidas e antibiofilme estavam principalmente associadas à presença de monoterpenos e fenilpropanóides. Este é o primeiro relato de óleos essenciais com potencial para controlar a contaminação ambiental e infecção animal por A. pleuropneumoniae, representando uma alternativa contra o crescente aumento de resistência bacteriana aos antibióticos.

Trypanosoma evansi in horses from Colombia - associated infection factors

Background: Trypanosoma evansi is the most common protozoan in tropical and subtropical regions of the world, due to its ability to maintain and be transmitted by vectors such as Stomoxys spp. and Tabanus spp. This protozoan causes high morbidity and mortality rates in horses in African, American and Asian countries. In the years 2021 and 2022, a high mortality rate was reported among horses with symptoms associated with Trypanosoma spp. in the municipality of Arauca, department of Arauca, Colombia. The investigation described here was therefore carried out, seeking to identify the pathogens and risk factors that led to the death of the horses in this region of Colombia. Cases: Blood samples were collected from Colombian criollo horses and dogs, as were samples of ticks, flies and horseflies that infested the horses. A variety of tissue samples were removed from the horses a few min after their death for histopathological analysis. Two questionnaires were applied to obtain information about the horses and the environment in which they live. The results of the clinical examination revealed pale mucous membranes, jaundice, high fever, dehydration and lethargy. The horses were also infested with Amblyomma mixtum (17.6%) and Dermacentor nitens (82.4%) ticks, and with Tabanus pungens (74%), Tabanus spp. (26%), and Stomoxys calcitrans flies (100%), while the dogs were infested with Rhipicephalus sanguineus s.l. (77.7%) and Amblyomma mixtum (22.2%) ticks. The blood smear test results revealed the presence of Trypanosoma spp. in 66.6% (n = 4) of the horse blood samples, and in 50% (n = 1) of the dog blood samples. PCR performed to identify the Trypanosoma species confirmed the presence of T. evansi. Histological examination of the spleen revealed the involvement and dissemination of T. evansi in the tissues. The horses also showed the presence of Equine Infectious Anemia Virus (EIAV). Discussion: This is the first updated specific report of T. evansi in criollo horses in the savannah flood zone of the municipality of Arauca, Colombia. The main risk associated with T. evansi infection in horses was found to be infestation with the natural vector T. pungens and the mechanical vector S. calcitrans, which are efficient ectoparasites for the transmission of this parasite. The presence of T. evansi in dogs represents a constant risk to horses, because dogs may serve as a reservoir for the maintenance of the hemoparasite in the population under study. Another risk factor for horses could be the presence of vampire bats (Desmodus rotundus), a species of bat that has been described as a vector and reservoir of T. evansi in Colombia. The presence of EIAV antibodies in the horses under study can be attributed to the exposure of sick horses to vectors of this virus, such as Tabanus spp., S. calcitrans and inanimate needle-shaped fomites. This is the first study that identifies the coinfection of T. evansi and EIAV in horses in the floodplain region of Colombia. In view of the importance of these 2 pathologies to the health of horses, a greater number of tests and a larger animal population will be required to determine if this coinfection is the cause of the death of criollo horses in this region of Colombia. Lastly, the owners reported that pharmacological control with trypanocides has not been successful in most of the outbreaks that occurred during the years 2021 and 2022. This may suggest that Trypanosoma evansi is developing resistance to these drugs; therefore, specific studies will be required in the future to test this hypothesis.

Alterations in blood glucose concentration in wild rodents, Holochilus sciureus, naturally infected with Schistosoma mansoni / Alterações na concentração de glicose do sangue periférico de roedores silvestres, Holochilus sciureus, naturalmente infectados com Schistosoma mansoni

The present study aimed to evaluate the changes in peripheral blood glucose concentrations induced by Schistosoma mansoni infection in Holochilus sciureus rodents, a wild reservoir of the parasite. Glucose concentration was measured in the plasma of blood samples using a colorimetric enzymatic test. Biological parameters and S. mansoni burden in each rodent were also verified and correlated with glucose concentrations. A total of 76 H. sciureus were captured, out of which 20 (26%) were infected with S. mansoni (n=13 males and n=7 females). Although the parasite burden was comparable between the sexes, blood glucose concentration was lower in infected males and almost unchanged in females. Furthermore, histopathological data revealed that male rodents had a greater hepatic granulomatous inflammatory reaction than females. In addition, we also confirmed that the weight and total length of the analyzed animals had no effect on glucose levels. Therefore, natural infection with S. mansoni in H. sciureus may have a lower impact on glycemic homeostasis in females, which will help us understand the role of these rodents as reservoirs of S. mansoni.(AU)

Neste presente estudo, objetivou-se avaliar as alterações induzidas pela infecção natural de Schistosoma mansoni nas concentrações de glicose do sangue periférico de roedores Holochilus sciureus, reservatório silvestre do parasito. A dosagem da concentração de glicose foi realizada no plasma de amostras de sangue por meio de teste enzimático colorimétrico. Parâmetros biológicos e carga parasitária de S. mansoni também foram verificados em cada roedor e correlacionados com as concentrações de glicose. Um total de 76 H. sciureus foram capturados, dentre os quais 20 (26%) estavam infectados por S. mansoni (n=13 machos e n=7 fêmeas). Apesar da carga parasitária ter sido semelhante entre os sexos, as concentrações de glicose no sangue foram menores em machos naturalmente infectados e quase inalteradas em fêmeas. Além disso, os dados histopatológicos mostraram uma maior reação inflamatória granulomatosa hepática em roedores machos, quando comparados com as fêmeas. Adicionalmente, também foi confirmado que o peso e o comprimento total dos animas analisados não interferiram nos valores de glicose. Portanto, a infecção natural por S. mansoni em H. sciureus pode induzir menos impacto na homeostase glicêmica em fêmeas, o que pode ajudar a melhor entender o papel desses roedores como reservatórios de S. mansoni.(AU)

Natural infection of Trypanosoma sp. in domestic sheep from Ecuador / Infecção natural do Trypanosoma sp. em ovelhas domésticas do Equador

Trypanosomosis in sheep is a hemoparasitic disease of worldwide interest due to its effects on the health of animals and the economic impact on producers, is caused by the protozoan Trypanosoma sp. In the present study, the occurrence of Trypanosoma sp. in sheep from the urban-rural area of the canton Salitre was determined, through an applied study with a qualitative descriptive prospective cross-sectional approach, carried out between October 1, 2019, and February 19, 2020. Blood samples were extracted from the jugular vein and analyzed by the blood smear method using the Giemsa and Diff-Quick staining techniques. Previously, in a first study realized between 2018 and 2019, 2 cases (2%) of Trypanosoma sp., 1 case of Babesia sp. (1%), and 4 cases of Anaplasma marginale (4%) were identified, but without the presence of symptoms of the disease. However, in this new research study, of 170 animals sampled from 5 herds and aged between 3 and 8 years, 34 (20%) were positive for Trypanosoma sp., 6 for Babesia sp. (3.52%) and 6 for A. marginale (3.52%) with coinfection between them. In this research, deteriorated clinical aspects and low hematological values were also determined in positive animals; besides of the total of positive cases, 25 presented symptoms of hemoparasitic disease, 3 sheep had abortions and 8 died. The results of this study showed that Trypanosoma sp. could already become an endemic parasitosis in sheep in the country, representing a serious problem of animal health.

A tripanossomose em ovinos é uma doença hemoparasitária de interesse mundial devido a seus efeitos sobre a saúde dos animais e o impacto econômico sobre os produtores, Esta doença é causada pelo protozoário Trypanosoma sp. No presente estudo, foi determinada a ocorrência de Trypanosoma sp. em ovelhas da área urbano-rural do Cantão Salitre, através de um estudo aplicado com uma abordagem transversal descritiva qualitativa prospectiva, realizado entre dia primeiro de outubro de 2019, à 19 de fevereiro de 2020. Amostras de sangue foram extraídas da veia jugular e analisadas pelo método de esfregaço de sangue usando as técnicas de coloração Giemsa e Diff-Quick. Anteriormente, em um primeiro estudo realizado entre 2018 e 2019, dois casos (2%) de Trypanosoma sp., foram identificados um caso de Babesia sp. (1%), e quatro casos de Anaplasma marginale (4%), mas sem a presença de sintomas da doença. Entretanto, neste novo estudo de pesquisa, de 170 animais amostrados de cinco rebanhos e com idades entre três e oito anos, 34 (20%) foram positivos para Trypanosoma sp., seis para Babesia sp. (3,52%) e seis para A. marginale (3,52%) com coinfecção entre eles. Nesta pesquisa, aspectos clínicos deteriorados e baixos valores hematológicos também foram determinados em animais positivos; além do total de casos positivos, 25 apresentaram sintomas de doença hemoparasítica, três ovelhas tiveram abortos e oito morreram. Os resultados deste estudo mostram que o Trypanosoma sp. já poderia se tornar uma parasitose endêmica em ovinos no país, representando um grave problema de saúde animal.