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Probiotics are live microorganisms that, when administered in adequate quantities, provide health benefits to the host. In this study, phenotypic and genotypic methods were used to evaluate the probiotic properties of Bacillus altitudinis 1.4. The isolate was sensitive to all antimicrobials tested and presented a positive result in the hemolysis test. B. altitudinis 1.4 spores were more resistant than vegetative cells, when evaluated in simulation of cell viability in the gastrointestinal tract, as well as adhesion to the intestinal mucosa. The isolate was capable of self-aggregation and coaggregation with pathogens such as Escherichia coli ATCC 25922 and Salmonella Enteritidis ATCC 13076. Genomic analysis revealed the presence of genes with probiotic characteristics. From this study it was possible to evaluate the gene expression of pro-inflammatory and anti-inflammatory cytokines for different treatments. Viable vegetative cells of B. altitudinis 1.4 increased the transcription of pro-inflammatory factors, in addition to also increasing the transcription of IL-10, indicating a tendency to stimulate a pro-inflammatory profile. Given the results presented, B. altitudinis 1.4 showed potential to be applied in the incorporation of this microorganism into animal feed, since the spores could tolerate the feed handling and pelletization processes.

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Trichomoniasis is a sexually transmitted infection caused by the protozoan Trichomonas vaginalis. Currently, trichomoniasis is treated with the class of nitroimidazoles, namely, metronidazole; however, resistant isolates and strains have been reported. The compounds derived from benzofuroxan are biologically active heterocycles. This study evaluated the in vitro antiparasitic activity of these compounds in trophozoites of T. vaginalis and determined the mean inhibitory concentration (IC50), minimum inhibitory concentration (MIC), mortality curve, and cytotoxicity. The compounds were named EH1, EH2, EH3, and EA2 and tested in various concentrations: 100 to 15 µM (EH1 and EH2); 100 to 5 µM (EH3); and 100 to 25 µM (EA2), respectively. The greatest efficacy was observed in the highest concentrations in 24 h, with inhibition of approximately 100% of trophozoites. Compounds EH2 and EH3 had the lowest MIC: EH2 (35 µM) and EH3 (45 µM), with IC50 of 11.33 µM and 6.83 µM, respectively. Compound EA2 was effective at the highest concentrations. The activity of the compounds in T. vaginalis started in the first hour of incubation with 90% inhibition; after 12 h, inhibition >95% was observed. Compound EH1 showed the lowest activity, with the highest activity between 12 and 24 h after incubation. These results demonstrate that benzofuroxan derivatives are promising compounds for the in vitro treatment of T. vaginalis.

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Seroprevalence of bovine alphaherpesvirus type 1 (BoAHV1) infections may be contaminated by crossreactive antibodies to bovine alphaherpesvirus type 5 (BoAHV5). To avoid such crossreactivity, an indirect enzyme-linked immunosorbent assay prepared with a recombinant glycoprotein C (gC) antigen (ELISA-gC1) was developed, aiming the detection of antibodies to BoAHV1, with no crossreactivity with BoAHV5 antibodies. The antigen for the ELISA-gC1 was the product of the expression of 219 bp from the N-terminal portion of the BoAHV1 gC gene, which bears low homology between the two virus types. The test was validated on 131 bovine serum samples, including 26 sera from BoAHV1-experimentally immunized, 38 sera from BoAHV5-experimentally infected or immunized calves, and 67 sera from calves seronegative for both BoAHV1 and BoAHV5, as determined by serum neutralization (SN). When compared to SN for BoAHV1, the ELISA-gC1 presented 100% sensitivity, 95.5 % specificity, 100 % negative predictive value, 89.6 % positive predictive value, 98.8 % precision, and a kappa correlation coefficient (κ) 0.95. None of the 38 BoAHV5-seropositive calves was detected by the ELISA-gC1. The ELISA-gC1 proved highly effective for the identification of BoAHV1-positive sera, with no crossreactivity with anti-BoAHV5 antibodies, thus able to distinguish serological responses from BoAHV1- and BoAHV5-seropositive cattle. Its capacity to detect BoAHV1-specific antibodies should allow the determination of the actual BoAHV1 prevalence in herds, which cannot be serologically determined in countries where BoAHV5 is also prevalent due to antibody crossreactivity. Apart from recognizing exclusively BoAHV1-infected cattle, the ELISA-gC1 may also be used in support of BoAHV5 epidemiological studies by allowing the exclusion of BoAHV1-seropositive animals.

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The present study was carried out to evaluate the intravaginal vaccine potential against bovine alphaherpesvirus type 5 (BoHV-5). Sixty three cows were divided into seven groups (n: 9) and inoculated intravaginally (VA) or intramuscularly (IM) with inactivated BoHV-5, associated with the recombinant B subunit of the heat-labile enterotoxin of E. coli (rLTB), 2-hydroxyethylcellulose (Drug Delivery System A - DDS-A) or Poloxamer 407 (Drug Delivery System B - DDS-B) as follows: G1 (DDS-A + BoHV-5 + rLTB), G2 (DDS-A + BoHV-5), G3 (DDS-B + BoHV-5 + rLTB), G4 (DDS-B + BoHV-5), G5 (BoHV-5 + rLTB), G6 (Negative control) e G7 (Positive control). The local and systemic humoral responses were measured by indirect ELISA (IgA and IgG) and serum neutralization tests, and the cellular response was measured by a quantitative direct ELISA (IL-2 and IFN-Gamma). The results showed the group inoculated by the IM route, G5, demonstrated the highest levels of IgG in the vaginal mucosa among the experimental groups (p < 0.05). In the groups tested with polymers (G1 and G3) in the vaginal mucosa, even higher levels of IgG were seen in comparison to the positive control (G7; p < 0.01). Higher levels of IgA were also noted in relation to the other groups (p < 0.05) on days 30, 60 and 90 post-inoculations. The groups G1 and G3 also provided higher titers of neutralizing antibodies (Log2) in relation to other treatments (p < 0.01) 90 days after inoculation. In the nasal mucosa, there was an increase in the levels of IgA and IgG with the use of vaccines from groups G1 and G3, in relation to the positive control, G7 (p < 0.05) at 60 and 90 days after the first inoculation. Moreover, neutralizing antibodies titers were detected at 60 and 90 days by serum neutralization. The inclusion of the evaluated polymers resulted in a superior response (p < 0.05) of immunoglobulins and IL-2 and IFN-Gamma in relation to the treatment using only rLTB (G5). This data demonstrates the capabilities of a vaccine with an intravaginal application in cattle to stimulate a local and systemic immune response.

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Fasciolosis is one of the most important parasitic diseases in animal health, affecting mainly ruminants, causing economic and productivity losses. This study aimed to evaluate the in vitro ovicidal and adulticidal activity of essential oils (EOs) from Pelargonium graveolens (geranium) and Citrus aurantium (sour orange) on Fasciola hepatica. Performed Gas Chromatography of EOs P. graveolens and C. aurantium, with major compound, citronellol (31.37 %) and limonene (93.89 %), respectively. For the cytotoxicity assay, the sour orange EO showed to be promising when used in lower concentrations. For the ovicidal tests, the eggs were incubated with geranium EOs at concentrations from 4.5 to 0.03375 mg/mL and sour orange at concentrations from 4.25 to 0.031875 mg/mL, along with controls. The viable eggs were counted on the 14th day post-incubation. Adult forms of F. hepatica were incubated containing the EOs and observed for 24 h after treatment, as well as the control groups. Later the specimens were fixed for histological analysis. Geranium and sour orange EOs in trematode eggs at the concentrations tested were 100 % effective in inactivating hatching (p < 0.05) when compared to the untreated control. In the adulticidal test, the essential oil of P. graveolens at both concentrations tested (0.0675 and 0.03375) within 15 h, promoted the death of flukes. For C. aurantium, 18 h was enough to inactivate all specimens, up to a concentration of 0.06375. The histological analysis, observed the accumulation of liquid in the tegument in the specimens incubated in C. aurantium and P. graveolens, with vacuolization in the tegument and spines, preventing externalization. The results of the study present OEs with efficient ovicidal and adulticidal activity.

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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.

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Felid alphaherpesvirus type 1 (FHV-1) is an important cause of respiratory and ocular diseases in cats worldwide. Mice have been widely used to study the pathogenesis of several human and animal viruses, especially herpesviruses. This study aimed to verify whether BALB/c mice are susceptible to FHV-1 infection. The animals were intranasally inoculated with FHV-1 and their clinical signs were observed from 3 days postinfection (dpi). At 10 dpi, the animals were euthanized and the lungs, liver, spleen, and kidneys were collected for histopathological examination and quantitative polymerase chain reaction. The results showed that mice were infected with FHV-1 and reproduced several features of the disease observed in its natural host. Histological lesions and viral DNA were found in all sampled tissues, with a higher frequency of FHV-1 DNA copies detected in the lungs. All mice were seroconverted to FHV-1 at 7 dpi. To our knowledge, this is the first report of experimental infection of BALB/c mice with FHV-1. Our findings demonstrate that this murine model can contribute to understanding of FHV-1 pathogenesis and may be useful for trials against this virus.(AU)

O herpesvírus felino tipo 1 (FHV-1) é um importante agente causador de doença respiratória e ocular em gatos em todo o mundo. Camundongos têm sido amplamente utilizados para estudar a patogenia de diversos vírus humanos e animais, especialmente os herpesvírus. O objetivo deste estudo foi verificar se camundongos BALB/c poderiam ser suscetíveis a infecção pelo FHV-1. Os animais foram inoculados com FHV-1 intranasalmente e sinais clínicos foram observados a partir do dia 3 após a infecção (dpi). Após dez dias da inoculação, os animais foram eutanasiados e os pulmões, fígado, baço e rins foram coletados para exame histopatológico e PCR quantitativo (qPCR). Os resultados mostraram que os camundongos foram infectados com o FHV-1 e reproduziram várias características da doença que são observadas em seu hospedeiro natural. Lesões histológicas e DNA viral foram detectadas em todos os tecidos amostrados, com maior frequência de DNA do FHV-1 nos pulmões. Todos os camundongos soroconverteram para FHV-1 aos 7 dpi. Para nosso conhecimento, este estudo é o primeiro relato de infecção experimental de camundongos BALB/c com FHV-1. Nossos resultados demonstraram que esse camundongo pode contribuir para o entendimento da patogenia do FHV-1 e pode ser um modelo útil para ensaios contra esse vírus.(AU)

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In worldwide there are reports of a significant decrease in colonies of the species Apis mellifera, caused by several factors, including viral infections. In order to study and diagnose illnesses caused by viruses, in vitro cell culture is used as a valuable tool. Yet, there are still no immortalized cell lines of honey bee Apis mellifera. Primary cell cultures are promising for this purpose and can supply the lack of continuous strains, but their establishment is difficult and laborious, which often makes them unfeasible for many research centers. Through the use of cell immortalization techniques, it is possible to develop continuous cell lines and thus benefit, in different ways, research related to different species of bees. The choice of technique is challenging, since in addition to the ability to remain viable for countless passages, cells must keep the genotype and phenotype similar or identical to the original tissue. This review intends to present methodologies that can be used to immortalize Apis mellifera cells, aiming to establish a cell line. The genotypic and phenotypic implications of each technique are evaluated, and the purpose of the cell line to be developed.(AU)

Ao redor do mundo há relatos da diminuição significativa de colônias da espécie Apis mellifera, causada por diversos fatores, incluindo infecções virais. Para estudo e diagnóstico de enfermidades causadas por vírus utiliza-se, como uma ferramenta valiosa, o cultivo celular in vitro. Contudo, ainda não existem linhagens celulares imortalizadas de abelhas Apis mellifera. Os cultivos celulares primários são promissores para este fim e podem suprir a falta de linhagens contínuas, porém seu estabelecimento é difícil e laborioso o que, muitas vezes, os torna inviáveis para muitos centros de pesquisa. Através do uso de técnicas de imortalização celular é possível desenvolver linhagens contínuas de células e assim beneficiar, de diversas formas, as pesquisas relacionadas às diferentes espécies de abelhas. A escolha da técnica é desafiadora, visto que, além da capacidade de permanecer viável por inúmeras passagens, as células devem manter o genótipo e fenótipo semelhante ou idêntico ao tecido original. O objetivo deste trabalho é apresentar metodologias que podem ser utilizadas para imortalização de células de Apis mellifera, visando o estabelecimento de uma linhagem celular. São avaliadas as implicações genotípicas e fenotípicas de cada técnica, e a finalidade da linhagem celular a ser desenvolvida.(AU)

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In worldwide there are reports of a significant decrease in colonies of the species Apis mellifera, caused by several factors, including viral infections. In order to study and diagnose illnesses caused by viruses, in vitro cell culture is used as a valuable tool. Yet, there are still no immortalized cell lines of honey bee Apis mellifera. Primary cell cultures are promising for this purpose and can supply the lack of continuous strains, but their establishment is difficult and laborious, which often makes them unfeasible for many research centers. Through the use of cell immortalization techniques, it is possible to develop continuous cell lines and thus benefit, in different ways, research related to different species of bees. The choice of technique is challenging, since in addition to the ability to remain viable for countless passages, cells must keep the genotype and phenotype similar or identical to the original tissue. This review intends to present methodologies that can be used to immortalize Apis mellifera cells, aiming to establish a cell line. The genotypic and phenotypic implications of each technique are evaluated, and the purpose of the cell line to be developed.

Ao redor do mundo há relatos da diminuição significativa de colônias da espécie Apis mellifera, causada por diversos fatores, incluindo infecções virais. Para estudo e diagnóstico de enfermidades causadas por vírus utiliza-se, como uma ferramenta valiosa, o cultivo celular in vitro. Contudo, ainda não existem linhagens celulares imortalizadas de abelhas Apis mellifera. Os cultivos celulares primários são promissores para este fim e podem suprir a falta de linhagens contínuas, porém seu estabelecimento é difícil e laborioso o que, muitas vezes, os torna inviáveis para muitos centros de pesquisa. Através do uso de técnicas de imortalização celular é possível desenvolver linhagens contínuas de células e assim beneficiar, de diversas formas, as pesquisas relacionadas às diferentes espécies de abelhas. A escolha da técnica é desafiadora, visto que, além da capacidade de permanecer viável por inúmeras passagens, as células devem manter o genótipo e fenótipo semelhante ou idêntico ao tecido original. O objetivo deste trabalho é apresentar metodologias que podem ser utilizadas para imortalização de células de Apis mellifera, visando o estabelecimento de uma linhagem celular. São avaliadas as implicações genotípicas e fenotípicas de cada técnica, e a finalidade da linhagem celular a ser desenvolvida.

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ABSTRACT: In worldwide there are reports of a significant decrease in colonies of the species Apis mellifera, caused by several factors, including viral infections. In order to study and diagnose illnesses caused by viruses, in vitro cell culture is used as a valuable tool. Yet, there are still no immortalized cell lines of honey bee Apis mellifera. Primary cell cultures are promising for this purpose and can supply the lack of continuous strains, but their establishment is difficult and laborious, which often makes them unfeasible for many research centers. Through the use of cell immortalization techniques, it is possible to develop continuous cell lines and thus benefit, in different ways, research related to different species of bees. The choice of technique is challenging, since in addition to the ability to remain viable for countless passages, cells must keep the genotype and phenotype similar or identical to the original tissue. This review intends to present methodologies that can be used to immortalize Apis mellifera cells, aiming to establish a cell line. The genotypic and phenotypic implications of each technique are evaluated, and the purpose of the cell line to be developed.

RESUMO: Ao redor do mundo há relatos da diminuição significativa de colônias da espécie Apis mellifera, causada por diversos fatores, incluindo infecções virais. Para estudo e diagnóstico de enfermidades causadas por vírus utiliza-se, como uma ferramenta valiosa, o cultivo celular in vitro. Contudo, ainda não existem linhagens celulares imortalizadas de abelhas Apis mellifera. Os cultivos celulares primários são promissores para este fim e podem suprir a falta de linhagens contínuas, porém seu estabelecimento é difícil e laborioso o que, muitas vezes, os torna inviáveis para muitos centros de pesquisa. Através do uso de técnicas de imortalização celular é possível desenvolver linhagens contínuas de células e assim beneficiar, de diversas formas, as pesquisas relacionadas às diferentes espécies de abelhas. A escolha da técnica é desafiadora, visto que, além da capacidade de permanecer viável por inúmeras passagens, as células devem manter o genótipo e fenótipo semelhante ou idêntico ao tecido original. O objetivo deste trabalho é apresentar metodologias que podem ser utilizadas para imortalização de células de Apis mellifera, visando o estabelecimento de uma linhagem celular. São avaliadas as implicações genotípicas e fenotípicas de cada técnica, e a finalidade da linhagem celular a ser desenvolvida.

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The white-eared opossums (Didelphis albiventris) is the largest Brazilian marsupial and a great example of animal synanthropy. Considering the high potential as a carrier of viruses originating from environmental contamination, the presence of Human adenovirus (AdV) and rotavirus was investigated in the feces of rescued white-eared opossums, which were in the process of rehabilitation. The feces of 49 animals were initially investigated by immunochromatography, with three samples positive for AdV and one sample positive for rotavirus. When submitted to PCR and nested PCR, the samples of six animals were positive for AdV and three animals were positive for group A rotavirus. Two positive samples in the immunochromatographic assay were not confirmed by PCR. After sequencing and phylogenetic analysis of AdV samples, all were identified within the genus Mastadenovirus, one being HAdV-C, four HAdV-E, and one being similar to a Mastadenovirus found in primates. This is the first report of molecular confirmation of human adenovirus and rotavirus in white-eared opossums. These data could be important of anticipation some emerging diseases and their effects on ecosystems health.

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Bacillus toyonensis is a probiotic microorganism that for decades has been used in animal nutrition around the world. The objective of this work was to evaluate the immunomodulatory effect of oral B. toyonensis supplementation in dogs vaccinated against canine parvovirus. Puppies were randomly selected and divided in two groups, one received B. toyonensis at a concentration of 2x10 8 viable spores per day and another group without supplementation was left as control. The puppies were vaccinated against canine parvovirus type 2. B. toyonensis supplementation was efficient in stimulating specific IgG for parvovirus with titers of 2, 3, and 2.5-fold higher than controls at 7, 21, and 35 pos-vaccination days respectively. Peripheral blood mononuclear cells (PBMCs) from dogs were cultured and stimulated with B. toyonensis DNA, vegetative cell and spores. The mRNA transcription of cytokines IL-4, IL-17, and IFN-γ up modulated by the stimuli. Thus, we conclude in this study that B. toyonensis supplementation may amplify the vaccine immune response against canine parvovirus.(AU)

Bacillus toyonensis é um micro-organismo probiótico que há décadas é utilizado na nutrição animal em todo o mundo. O objetivo deste trabalho foi avaliar o efeito imunomodulador da suplementação oral de B. toyonensis em cães vacinados contra o parvovírus canino. Os filhotes foram selecionados aleatoriamente e divididos em dois grupos, um recebeu B. toyonensis na concentração de 2 × 10 8 esporos viáveis por dia e outro grupo sem suplementação como controle. Os filhotes foram vacinados contra o parvovírus canino tipo 2. A suplementação com B. toyonensis foi eficiente em estimular IgG específica para parvovírus com títulos de 2, 3 e 2,5 vezes maior que os controles aos 7, 21 e 35 dias pós-vacinação, respectivamente. Células mononucleares do sangue periférico (PBMCs) de cães foram cultivadas e estimuladas com DNA de B. toyonensis, células vegetativas e esporos. A transcrição do mRNA das citocinas IL-4, IL-17 e IFN-γ foi modulada pelos estímulos. Assim, concluímos neste estudo que a suplementação com B. toyonensis pode amplificar a resposta imune da vacina contra o parvovírus canino.(AU)

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Bees are fundamental in several aspects, especially in relation to plant biodiversity and pollination. Recently, immense losses are being faced in the number of Brazilian colonies, mainly in southern states of the country, which has a strong beekeeping activity. There are indications that, among the reasons for the losses, pathogens that affect the health of bees may be involved. Among them, the microsporidium Nosema and the black queen cell virus (BQCV) stand out for their prevalence. In this study, 92 colonies of 17 apiaries from southern Brazil were evaluated for infection by Nosema ceranae, Nosema apis and BQCV. Nucleic acid extractions and cDNA synthesis were performed from adult bee samples, followed by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and multiplex PCR. Eight BQCV positive samples were subjected to sequencing. The results showed that N. ceranae and BQCV are circulating in the Southern region of the country, which may be the reason for the loss of colonies. N. apis was not found. N. ceranae was found in 57.6% (53/92) of the colonies and BQCV in 32.6% (30/92). Co-infection was found in 25% (23/92) of the colonies studied, a factor that is suggested to be reducing the hosts' longevity due to the synergistic action of the pathogens. The samples submitted to sequencing indicated similarity of 96.8 to 100% between them, in addition to strong similarity with sequences from Asia, United States, Germany and Peru. This study reports the circulation of N. ceranae and BQCV in apiaries in southern Brazil, in addition to being the first phylogenetic analysis of the Brazilian BQCV sequence.(AU)

As abelhas mostram-se fundamentais em diversos aspectos, especialmente com relação à biodiversidade de plantas e polinização. Recentemente, estão sendo enfrentadas imensas perdas no número de colônias brasileiras, principalmente nos estados do sul do país, com forte atividade apícola. Há indicativos de que, dentre as razões para as perdas, possam estar envolvidos patógenos que afetam a saúde das abelhas. Dentre eles, o microsporídio Nosema e o vírus da realeira negra (BQCV) destacam-se pela prevalência. Neste estudo, foram avaliadas 92 colônias, de 17 apiários do sul do Brasil, a respeito da infecção por Nosema ceranae, Nosema apis e BQCV. Foram realizadas extrações de ácidos nucleicos e síntese de cDNA a partir de amostras de abelhas adultas, seguidos de Reação em Cadeia da Polimerase-Transcriptase Reversa (RT-PCR). Oito amostras positivas para BQCV foram submetidas a sequenciamento. Os resultados mostraram que N. ceranae e BQCV estão circulando na região sul do país, podendo ser a razão para as perdas de colônias. N. apis não foi encontrado. N. ceranae foi encontrado em 57.6% (53/92) das colônias e BQCV em 32.6% (30/92). Foi encontrada coinfecção por ambos em 25% (23/92) das colônias estudadas, fator que sugere a diminuição da longevidade do hospedeiro por ação sinérgica dos patógenos. As amostras submetidas ao sequenciamento indicaram similaridade de 96.8 a 100% entre elas, além de forte similaridade com sequências da Ásia, Estados Unidos, Alemanha e Peru. Este estudo relata a circulação de N. ceranae e BQCV nos apiários do sul do Brasil, além de ser a primeira análise filogenética da sequência do BQCV brasileiro.(AU)

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Bacillus toyonensis is a probiotic microorganism that for decades has been used in animal nutrition around the world. The objective of this work was to evaluate the immunomodulatory effect of oral B. toyonensis supplementation in dogs vaccinated against canine parvovirus. Puppies were randomly selected and divided in two groups, one received B. toyonensis at a concentration of 2x10 8 viable spores per day and another group without supplementation was left as control. The puppies were vaccinated against canine parvovirus type 2. B. toyonensis supplementation was efficient in stimulating specific IgG for parvovirus with titers of 2, 3, and 2.5-fold higher than controls at 7, 21, and 35 pos-vaccination days respectively. Peripheral blood mononuclear cells (PBMCs) from dogs were cultured and stimulated with B. toyonensis DNA, vegetative cell and spores. The mRNA transcription of cytokines IL-4, IL-17, and IFN-γ up modulated by the stimuli. Thus, we conclude in this study that B. toyonensis supplementation may amplify the vaccine immune response against canine parvovirus.(AU)

Bacillus toyonensis é um micro-organismo probiótico que há décadas é utilizado na nutrição animal em todo o mundo. O objetivo deste trabalho foi avaliar o efeito imunomodulador da suplementação oral de B. toyonensis em cães vacinados contra o parvovírus canino. Os filhotes foram selecionados aleatoriamente e divididos em dois grupos, um recebeu B. toyonensis na concentração de 2 × 10 8 esporos viáveis por dia e outro grupo sem suplementação como controle. Os filhotes foram vacinados contra o parvovírus canino tipo 2. A suplementação com B. toyonensis foi eficiente em estimular IgG específica para parvovírus com títulos de 2, 3 e 2,5 vezes maior que os controles aos 7, 21 e 35 dias pós-vacinação, respectivamente. Células mononucleares do sangue periférico (PBMCs) de cães foram cultivadas e estimuladas com DNA de B. toyonensis, células vegetativas e esporos. A transcrição do mRNA das citocinas IL-4, IL-17 e IFN-γ foi modulada pelos estímulos. Assim, concluímos neste estudo que a suplementação com B. toyonensis pode amplificar a resposta imune da vacina contra o parvovírus canino.(AU)

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Bees are fundamental in several aspects, especially in relation to plant biodiversity and pollination. Recently, immense losses are being faced in the number of Brazilian colonies, mainly in southern states of the country, which has a strong beekeeping activity. There are indications that, among the reasons for the losses, pathogens that affect the health of bees may be involved. Among them, the microsporidium Nosema and the black queen cell virus (BQCV) stand out for their prevalence. In this study, 92 colonies of 17 apiaries from southern Brazil were evaluated for infection by Nosema ceranae, Nosema apis and BQCV. Nucleic acid extractions and cDNA synthesis were performed from adult bee samples, followed by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and multiplex PCR. Eight BQCV positive samples were subjected to sequencing. The results showed that N. ceranae and BQCV are circulating in the Southern region of the country, which may be the reason for the loss of colonies. N. apis was not found. N. ceranae was found in 57.6% (53/92) of the colonies and BQCV in 32.6% (30/92). Co-infection was found in 25% (23/92) of the colonies studied, a factor that is suggested to be reducing the hosts' longevity due to the synergistic action of the pathogens. The samples submitted to sequencing indicated similarity of 96.8 to 100% between them, in addition to strong similarity with sequences from Asia, United States, Germany and Peru. This study reports the circulation of N. ceranae and BQCV in apiaries in southern Brazil, in addition to being the first phylogenetic analysis of the Brazilian BQCV sequence.(AU)

As abelhas mostram-se fundamentais em diversos aspectos, especialmente com relação à biodiversidade de plantas e polinização. Recentemente, estão sendo enfrentadas imensas perdas no número de colônias brasileiras, principalmente nos estados do sul do país, com forte atividade apícola. Há indicativos de que, dentre as razões para as perdas, possam estar envolvidos patógenos que afetam a saúde das abelhas. Dentre eles, o microsporídio Nosema e o vírus da realeira negra (BQCV) destacam-se pela prevalência. Neste estudo, foram avaliadas 92 colônias, de 17 apiários do sul do Brasil, a respeito da infecção por Nosema ceranae, Nosema apis e BQCV. Foram realizadas extrações de ácidos nucleicos e síntese de cDNA a partir de amostras de abelhas adultas, seguidos de Reação em Cadeia da Polimerase-Transcriptase Reversa (RT-PCR). Oito amostras positivas para BQCV foram submetidas a sequenciamento. Os resultados mostraram que N. ceranae e BQCV estão circulando na região sul do país, podendo ser a razão para as perdas de colônias. N. apis não foi encontrado. N. ceranae foi encontrado em 57.6% (53/92) das colônias e BQCV em 32.6% (30/92). Foi encontrada coinfecção por ambos em 25% (23/92) das colônias estudadas, fator que sugere a diminuição da longevidade do hospedeiro por ação sinérgica dos patógenos. As amostras submetidas ao sequenciamento indicaram similaridade de 96.8 a 100% entre elas, além de forte similaridade com sequências da Ásia, Estados Unidos, Alemanha e Peru. Este estudo relata a circulação de N. ceranae e BQCV nos apiários do sul do Brasil, além de ser a primeira análise filogenética da sequência do BQCV brasileiro.(AU)

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Felid alphaherpesvirus 1 (FeHV-1) and feline calicivirus (FCV) affect cats worldwide. The aim of this study was to evaluate the frequency of occurrence of FeHV-1 and FCV in cats with clinical signs of respiratory, oral and/or ocular disease. Samples were collected from cats cared for in veterinary ambulatory and clinics and submitted to molecular detection and viral isolation. Of the 49 cats evaluated, 45 (92%) were positive for at least one of the viruses; 82% (40/49) were positive for FeHV-1 and 41% (20/49) for FCV. Of these, 31% (15/49) were coinfection cases. For FeHV-1, 45% (18/40) of the cats tested were positive from the collection of eye swab, and the same percentage (9/20) was obtained for the FCV by the oral swab. FeHV-1 and/or FCV were isolated in 35% (17/49) of the samples. The main clinical sign observed was ocular secretion in 71% (35/49) of cats, characterized as mild serous, purulent or serosanguineous, and in some cases associated with ocular injury and marked chemosis. Our findings demonstrate the high occurrence of FeHV-1 and FCV in domestic cats in southern Brazil and indicate that measures should be implemented to improve the diagnostic, prevention and management against of these important diseases.(AU)

Alphaherpesvírus felídeo 1 (FeHV-1) e calicivírus felino (FCV) afetam gatos mundialmente. O objetivo deste estudo foi identificar a frequência de ocorrência de FeHV-1 e FCV em gatos com sinais clínicos de doença respiratória, oral e/ou ocular. Amostras foram coletadas de gatos atendidos em ambulatório e clínicas veterinárias e submetidas à detecção molecular e isolamento viral. Dos 49 gatos avaliados, 45 (92%) foram positivos para ao menos um dos vírus; 82% (40/49) foram positivos para o FeHV-1 e 41% (20/49) para o FCV. Destes, 31% (15/49) foram casos de coinfecção. Para o FeHV-1, 45% (18/40) dos gatos foram positivos na coleta do swab ocular, e o mesmo percentual (9/20) foi obtido para o FCV a partir do swab oral. FeHV-1 e/ou FCV foram isolados em 35% (17/49) das amostras. O principal sinal clínico observado foi secreção ocular em 71% (35/49) dos gatos, caracterizada como serosa, purulenta ou serossanguinolenta e, em alguns casos, associada à lesão e quemose. Nossos resultados demonstram a alta ocorrência de FeHV-1 e FCV em gatos domésticos na região Sul do Brasil e indicam que devem ser implementadas medidas para melhorar o diagnóstico, a prevenção e o manejo contra essas importantes doenças.(AU)

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Felid alphaherpesvirus 1 (FeHV-1) and feline calicivirus (FCV) affect cats worldwide. The aim of this study was to evaluate the frequency of occurrence of FeHV-1 and FCV in cats with clinical signs of respiratory, oral and/or ocular disease. Samples were collected from cats cared for in veterinary ambulatory and clinics and submitted to molecular detection and viral isolation. Of the 49 cats evaluated, 45 (92%) were positive for at least one of the viruses; 82% (40/49) were positive for FeHV-1 and 41% (20/49) for FCV. Of these, 31% (15/49) were coinfection cases. For FeHV-1, 45% (18/40) of the cats tested were positive from the collection of eye swab, and the same percentage (9/20) was obtained for the FCV by the oral swab. FeHV-1 and/or FCV were isolated in 35% (17/49) of the samples. The main clinical sign observed was ocular secretion in 71% (35/49) of cats, characterized as mild serous, purulent or serosanguineous, and in some cases associated with ocular injury and marked chemosis. Our findings demonstrate the high occurrence of FeHV-1 and FCV in domestic cats in southern Brazil and indicate that measures should be implemented to improve the diagnostic, prevention and management against of these important diseases.(AU)

Alphaherpesvírus felídeo 1 (FeHV-1) e calicivírus felino (FCV) afetam gatos mundialmente. O objetivo deste estudo foi identificar a frequência de ocorrência de FeHV-1 e FCV em gatos com sinais clínicos de doença respiratória, oral e/ou ocular. Amostras foram coletadas de gatos atendidos em ambulatório e clínicas veterinárias e submetidas à detecção molecular e isolamento viral. Dos 49 gatos avaliados, 45 (92%) foram positivos para ao menos um dos vírus; 82% (40/49) foram positivos para o FeHV-1 e 41% (20/49) para o FCV. Destes, 31% (15/49) foram casos de coinfecção. Para o FeHV-1, 45% (18/40) dos gatos foram positivos na coleta do swab ocular, e o mesmo percentual (9/20) foi obtido para o FCV a partir do swab oral. FeHV-1 e/ou FCV foram isolados em 35% (17/49) das amostras. O principal sinal clínico observado foi secreção ocular em 71% (35/49) dos gatos, caracterizada como serosa, purulenta ou serossanguinolenta e, em alguns casos, associada à lesão e quemose. Nossos resultados demonstram a alta ocorrência de FeHV-1 e FCV em gatos domésticos na região Sul do Brasil e indicam que devem ser implementadas medidas para melhorar o diagnóstico, a prevenção e o manejo contra essas importantes doenças.(AU)

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Avian poxvirus (APV) is an enveloped double-stranded DNA virus that affects many domestic and wild birds worldwide. APVs are classified into three clades (A to C), represented by fowlpox (FP) virus (clade A), canarypox virus (clade B), and psittacinepox virus (clade C), although two additional clades (D and E) have been proposed. In this study, a tumorlike skin lesion found in a domestic fowl was submitted for molecular diagnosis of Avipoxvirus by PCR and sequencing. The phylogenetic analysis revealed that the amplified segment of the corelike 4b protein and polymerase genes clustered in clade E. The APVs in clade E were previously reported from outbreaks in Hungary (flock of turkeys) and in Mozambique (layer chickens), associated with a possible vaccine failure to protect against clade E viruses. To our knowledge, this report is the first identification of clade E in this country, providing new information about host range and genetic diversity of APVs in Brazil, and may represent a potential risk of FP disease outbreaks in commercial poultry.

Reporte de caso- Identificación del Avipoxvirus clado E en Brasil. El poxvirus aviar (APV) es un virus de ADN bicatenario envuelto que afecta a muchas aves domésticas y silvestres en todo el mundo. Los poxvirus aviares se clasifican en tres clados (A, B y C), representados por el virus de la viruela aviar (FP) (clado A), el virus de la viruela del canario (clado B) y el virus de la viruela de los psitácidos (clado C), aunque dos clados adicionales (D y E) han sido propuestos. En este estudio, una lesión cutánea similar a un tumor encontrada en una gallina doméstica fue sometida a diagnóstico molecular de Avipoxvirus por PCR y secuenciación. El análisis filogenético reveló que el segmento amplificado de los genes de la proteína del centro 4b y de la polimerasa se agruparon en el clado E. Los poxvirus aviares en el clado E se reportaron previamente de brotes en Hungría (parvada de pavos) y en Mozambique (gallinas de postura), asociados con una posible falla de la vacuna para proteger contra los virus del clado E. De acuerdo con el conocimiento de los autores, este informe es la primera identificación del clado E en este país, brindando nueva información sobre el rango de hospedadores y la diversidad genética de poxvirus aviares en Brasil, y puede representar un riesgo potencial de brotes de viruela aviar en aves comerciales.

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Pythium insidiosum belongs to the phylum Oomycota. It is capable of infecting mammals causing a serious condition called pythiosis, which affects mainly horses in Brazil and humans in Thailand. The objective of the present study was to verify the in vitro anti-P. insidiosum activity of a biogenic silver nanoparticle (bio-AgNP) formulation. The in vitro assays were evaluated on P. insidiosum isolates (n = 38) following the M38-A2 protocol. Damage to the P. insidiosum hyphae ultrastructure was verified by means of scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Bio-AgNP inhibition concentrations on P. insidiosum isolates ranged from 0.06 to 0.47 µg/ml. It was observed through SEM that P. insidiosum hyphae treated showed surface roughness, as well as cell walls with multiple retraction areas, loss of continuity, and rupture in some areas. The TEM of treated hyphae did not differentiate organelle structures; also, the cellular wall was rarefied, showing wrinkled and partly ruptured borders. The bio-AgNP evaluated has excellent in vitro anti-P. insidiosum activity. However, further studies on its in vivo action are necessary as so to determine the possibility of its use in the treatment of the disease in affected hosts.

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Background: The introduction of any infectious agent into an industrial or subsistence farm worries agribusiness owners in Brazilbecause it reduces product quality and increases treatment costs, although most diseases are untreatable, thus causing economic losseswith morbidity and mortality. Therefore, an epidemiological survey of viral diseases associated with poultry was developed by performing a detailed description of the risk factors that may be related to existing diseases using domestic poultry sample data recordedin the Regional Diagnostic Laboratory (LRD) of College of Veterinary Medicine of the Federal University of Pelotas (UFPel), RioGrande do Sul, Brazil, from 2000 to 2016.Materials, Methods & Results: Epidemiological and clinical-pathological data were collected and then compared with disease databy multivariate analysis using statistical EpiInfo version 6.04 and Microsoft Office Excel 2010 software. The frequencies and 95%confidence intervals (CI), association measures (odds ratio=OR and relative risk=RR), Chi-square test, and the results consideredsignificant with a value of P ≤ 0.05 were described. A total of 410 samples of domestic poultry were tested, and the results showed66 (16.1%) viral diseases. The following conditions were the most commonly found diseases in this study: Mareks disease (42.4%),Infectious bursal disease (31.8%), Avian leukosis (16.6%), Avian pox (7.5%) and Avian infectious bronchitis (1.5%). In this articlewe discuss the most frequent viral diseases: Mareks disease (DM) and Gumboro disease. It was also possible to conclude that birdswith Mareks disease presented higher odds of developing nerve, tegumentary and locomotors signs (P ≤ 0.05). As well as, morelikely to present tumoriform lesions in the liver, spleen, kidneys and heart P ≤ 0.05, as well as lesions in the proventriculus, musclelesions and in the sciatic nerve P ≤ 0.05. Laying poultry...(AU)

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