Resumo O gênero Dermocystidium infecta uma ampla variedade de espécies de animais, e o hospedeiro pode se infectar pela ingestão de endósporos. O diagnóstico depende de montagem em lâmina a fresco e análise histopatológica dos tecidos afetados. O objetivo deste estudo foi relatar infecção por Dermocystidium na pele do bagre listrado (P. hypophthalmus), cultivado em piscicultura do estado do Ceará, nordeste do Brasil. Foi detectado que 100% dos animais analisados estavam infectados por Dermocystidium sp. A montagem em lâminas a fresco e histopatologia das lesões mostrou cistos repletos de esporos entre a derme e a epiderme, encapsulados por tecidos conectivos. Devido às escassas pesquisas sobre esse parasito, em diferentes espécies de peixes no Brasil e no mundo, estudos adicionais são necessários para entender a endemicidade em diferentes pisciculturas do Brasil, consequentemente, melhorando os métodos de prevenção da doença e aumentando a produção e a produtividade.

Abstract The genus Dermocystidium infects a wide range of animals. The host infection often occurs through the ingestion of endospores. The diagnosis depends on wet mounts and histopathological analysis of the affected tissue. The aim of this study was to investigate the incidence of Dermocystidium sp. infection on the skin of farmed striped catfish (Pangasianodon hypophthalmus) from a fish farm located in Fortaleza, Ceará state, northeastern Brazil. From these observations, we determined that 100% of the analyzed animals were infected with Dermocystidium sp. The wet mount and histopathology of the fish lesions revealed spore-filled cysts between the dermis and epidermis, encapsulated by connective tissue. Owing to a lack of research on the parasite and its prevalence among different fish species in Brazil and the rest of the world, additional studies are required to understand their endemicity in fish farms of Brazil, and consequently develop better disease prevention methods and increase the overall productivity.

Resumo Scuticociliatose, causada pelo protozoário ciliado oportunista, é responsável por significativas perdas econômicas em peixes marinhos ornamentais. O estudo relata a ocorrência de Uronema spp., parasitando dez espécies de peixes de recife em um distribuidor de peixes ornamentais: “Blue green damselfish” (Chromis viridis), “Vanderbilt’s Chromis” (Chromis vanderbilti), “Pennant coralfish” (Heniochus acuminatus), “Threespot angelfish” (Apolemichthys trimaculatus), “Goldspotted angelfish” (Apolemichthys xanthopunctatus), “Sea goldie” (Pseudanthias squamipinnis), “Orchid dottyback” (Pseudochromis fridmani), “Threadfin butterflyfish” (Chaetodon auriga), “Vagabond butterflyfish” (Chaetodon vagabundus), e “Bluecheek butterflyfish” (Chaetodon semilarvatus). Peixes doentes apresentaram distúrbios como hemorragias e lesões ulcerativas na superfície do corpo. A análise histopatológica do músculo, fígado, intestino, rim, baço, brânquias e estômago revelou hemorragias e degeneração das fibras musculares, degeneração vacuolar de hepatócitos, processo inflamatório e granuloma no fígado, atrofia das vilosidades intestinais, processo inflamatório e granuloma no rim, centros de melanomacrófagos e processo inflamatório no baço, hiperplasia das células epiteliais, bem como formação de granuloma nas brânquias e degeneração vacuolar e eosinófilos no estômago. Devido à gravidade da doença, é necessário implementar medidas de biossegurança com diagnóstico rápido e preciso para minimizar o risco de perdas econômicas causadas por Uronema spp.

Abstract Scuticociliatosis, caused by an opportunistic ciliate protozoan, is responsible for significant economic losses in marine ornamental fish. This study reports the occurrence of Uronema spp., parasitizing ten species of marine reef fish at an ornamental fish wholesaler: Blue green damselfish (Chromis viridis), Vanderbilt’s Chromis (Chromis vanderbilti), Pennant coralfish (Heniochus acuminatus), Threespot angelfish (Apolemichthys trimaculatus), Goldspotted angelfish (Apolemichthys xanthopunctatus), Sea goldie (Pseudanthias squamipinnis), Orchid dottyback (Pseudochromis fridmani), Threadfin butterflyfish (Chaetodon auriga), Vagabond butterflyfish (Chaetodon vagabundus), and Bluecheek butterflyfish (Chaetodon semilarvatus). Diseased fish showed disorders such as hemorrhages and ulcerative lesions on the body surface. Histopathological analysis of the muscle, liver, gut, kidney, spleen, gills, and stomach revealed hemorrhages and degeneration of muscle fiber, vacuolar degeneration of hepatocyte, inflammatory process and granuloma in the liver, atrophy of intestinal villi, inflammatory process and granuloma in the kidney, melanomacrophage centers, as well as inflammatory process in the spleen, epithelial cells hyperplasia and granuloma formation in the gills, and vacuolar degeneration and eosinophils in the stomach. Due to the severity of the disease, it is necessary to implement biosecurity measures with rapid and accurate diagnosis to minimize the risk of economic losses caused by Uronema spp.

ABSTRACT Matrix Assisted Laser Desorption/Ionization and Time of Flight mass spectrometry (MALDI-TOF MS) is a powerful tool for the identification of bacteria through the detection and analysis of their proteins or fragments derived from ribosomes. Slight sequence variations in conserved ribosomal proteins distinguish microorganisms at the subspecies and strain levels. Characterization of Leptospira spp. by 16S RNA sequencing is costly and time-consuming, and recent studies have shown that closely related species (e.g., Leptospira interrogans and Leptospira kirschneri) may not be discriminated using this technology. Herein, we report an in-house Leptospira reference spectra database using Leptospira reference strains that were validated with a collection of well-identified Brazilian isolates kept in the Bacterial Zoonosis Laboratory at the Veterinary Preventive Medicine and Animal Health Department at Sao Paulo University. In addition, L. interrogans and L. kirschneri were differentiated using an in-depth mass spectrometry analysis with ClinProTools™ software. In conclusion, our in-house reference spectra database has the necessary accuracy to differentiate pathogenic and non-pathogenic species and to distinguish L. interrogans and L. kirschneri.

Cats are often described as carriers of Pasteurella multocida in their oral microbiota. This agent is thought to cause pneumonia, conjunctivitis, rhinitis, gingivostomatitis, abscess and osteonecrosis in cats. Human infection with P. multocida has been described in several cases affecting cat owners or after cat bites. In Brazil, the cat population is approximately 21 million animals and is increasing, but there are no studies of the presence of P. multocida in the feline population or of human cases of infection associated with cats. In this study, one hundred and ninety-one healthy cats from owners and shelters in São Paulo State, Brazil, were evaluated for the presence of P. multocida in their oral cavities. Twenty animals were positive for P. multocida, and forty-one strains were selected and characterized by means of biochemical tests and PCR. The P. multocida strains were tested for capsular type, virulence genes and resistance profile. A total of 75.6% (31/41) of isolates belonged to capsular type A, and 24.4% (10/41) of the isolates were untypeable. None of the strains harboured toxA, tbpA or pfhA genes. The frequencies of the other genes tested were variable, and the data generated were used to build a dendrogram showing the relatedness of strains, which were clustered according to origin. The most common resistance profile observed was against sulfizoxazole and trimethoprim-sulphamethoxazole.

The effects of phytogenic feed additives on piglet performance and fecal score (FDD), as well as on lipid oxidation of pork meat were evaluated. One hundred and twenty crossbred weaned piglets were randomly assigned to six treatments according to a 2 × 3 factorial design with five replicates per treatment. Factors were: challenge with Salmonella Typhimurium at 35 days of age or no challenge, and three different additives (control (CTR), basal diet; phytogenic feed additives (PHY), basal diet plus 2000 ppm of phytogenic feed additives - Rosmarinus officinalis, Mentha piperita, Lippia sidoides and Porophyllum ruderale; and antimicrobial agent (ATB), basal diet plus 100 ppm of tylosin, 2000 ppm of zinc and colistin sulfate, 30 ppm in the pre-starter basal diet, 10 ppm in the starter basal diet I and II, and 5 ppm in growth and finishing basal diet). Body weight (BW) of the piglets of ATB was greater throughout the experimental period, without any differences detected between CTR and PHY. Nevertheless, from 96 to 106 days of age, the BW of the CTR group was greater than PHY. From 21 to 34 days of age, feed conversion of ATB was lower than CTR; however, PHY showed an intermediate result, which did not differ from either ATB or CTR. Challenged animals reduced feed intake from day 35 to 48 compared with unchallenged animals. Piglet performance and fecal score from 21 to 48 days of age were lower in piglets that received ATB compared with the other treatments. However, from 35 to 48 days of age, the FDD of PHY was lower than CTR. Lipid oxidation was not reduced in treated animals. Antimicrobial agent improved the growth performance of piglets until 63 days of age, and no difference was observed between the treatments from 64 to 131 days of age. Antimicrobial agent reduced FDD; the FDD of PHY was similar to that of ATB after 48 days. None of the treatments affected lipid oxidation of pork meat.

Clostridium perfringens é uma bactéria Gram positiva anaeróbica, conhecida por infectar os seres humanos, animais domésticos e de vida selvagem. Apesar de as infecções causadas por C. perfringens tipo C e A em suínos serem bastante estudadas, poucos relatos descrevem a relação genética entre as linhagens envolvidas na cadeia epidemiológica da clostridiose suína, bem como a presença do microorganismo em abatedouros. O objetivo do presente estudo foi isolar C. perfringens a partir das fezes e carcaças de suínos no abatedouro, caracterizar os isolados quanto à presença dos genes codificadores de enterotoxina, toxina alfa, beta, épsilon, iota e beta 2 através da PCR e comparar os isolados através da eletroforese em campo pulsado (PFGE). A frequência de isolamento do agente em carcaças e em intestinos de suínos foi de 58,8% para ambos os tipos de amostras. De acordo com a reação em cadeia pela polimerase, somente a toxina alfa foi detectada, sendo todos os isolados negativos para toxina beta2 e enterotoxina. Através da técnica de PFGE, as cepas foram caracterizadas em 35 pulsotipos, sendo que, em apenas um caso, um isolado de amostras de carcaças foi agrupado no mesmo pulsotipo do isolado de fezes do mesmo animal, indicando que a possibilidade de contaminação cruzada no processamento da carcaça foi baixa. Apesar da alta prevalência de C. perfringens em carcaças de suínos provenientes dos abatedouros avaliados, o risco de intoxicação alimentar para os consumidores de carne suína brasileira é baixo, já que todas as cepas foram negativas para o gene cpe, codificador de enterotoxina.

Clostridium perfringens is an anaerobic Gram-positive bacterium known as common pathogen for humans, for domestic and wildlife animals. Although infections caused by C. perfringens type C and A in swine are well studied, just a few reports describe the genetic relationship among strains in the epidemiological chain of swine clostridioses, as well as the presence of the microorganism in the slaughterhouses. The aim of the present study was to isolate C. perfringens from feces and carcasses from swine slaughterhouses, characterize the strains in relation to the presence of enterotoxin, alpha, beta, epsilon, iota and beta-2 toxins genes, using polymerase chain reaction (PCR) and comparing strains by means of Pulsed field gel electrophoresis (PFGE). Clostridium perfringens isolation frequencies in carcasses and finishing pig intestines were of 58.8% in both types of samples. According to the polymerase chain reaction assay, only alfa toxin was detected, being all isolates also negative to enterotoxin and beta2 toxin. Through PFGE technique, the strains were characterized in 35 pulsotypes. In only one pulsotype, the isolate from carcass sample was grouped with fecal isolate of the same animal, suggesting that the risk of cross-contamination was low. Despite the high prevalence of C. perfringens in swine carcasses from the slaughterhouses assessed, the risk of food poisoning to Brazilian pork consumers is low, since all strains were negative to cpe-gene, codifying enterotoxin.

Haemophilus parasuis infection, known as Glässer's disease, is characterized by fibrinous polyserositis, arthritis and meningitis in piglets. Although traditional diagnosis is based on herd history, clinical signs, bacterial isolation and serotyping, the molecular-based methods are alternatives for species-specific tests and epidemiologic study. The aim of this study was to characterize H. parasuis strains isolated from different states of Brazil by serotyping, PCR and ERIC-PCR. Serotyping revealed serovar 4 as the most prevalent (24 %), followed by serovars 14 (14 %), 5 (12 %), 13 (8 %) and 2 (2 %), whereas 40 % of the strains were considered as non-typeable. From 50 strains tested 43 (86%) were positive to Group 1 vtaA gene that have been related to virulent strains of H.parasuis. ERIC-PCR was able to type isolates tested among 23 different patterns, including non-typeable strains. ERIC-PCR patterns were very heterogeneous and presented high similarity between strains of the same animal or farm origin. The results indicated ERIC-PCR as a valuable tool for typing H. parasuis isolates collected in Brazil.

Três ensaios comparativos foram feitos com o objetivo de aperfeiçoar a sensibilidade do diagnóstico da infecção pela Bordetella bronchiseptica a partir de suabes nasais de leitões. O experimento inicial comparou a recuperação de B. bronchiseptica a partir de suabes, simultaneamente inoculados com B. bronchiseptica e algumas bactérias interferentes, imersos em três formulações para transporte (meio Amies com carvão, caldo tripticaseína de soja e tampão de fosfatos segundo Soerensen suplementado com 5% de soro fetal bovino) e submetidos a diferentes temperaturas (10ºC e 27ºC) e períodos de incubação (24, 72 e 120 horas). O experimento subseqüente comparou três meios seletivos (ágar MacConkey, meio seletivo G20G modificado e o meio de ceftiofur) em relação às suas capacidades de recuperação a partir de amostras clínicas. O último experimento comparou a reação em cadeia pela polimerase aos três meios seletivos. No primeiro experimento, a recuperação de B. bronchiseptica nos sistemas de transporte foi melhor a 27ºC e as três formulações tiveram boas performances nesta temperatura, mas o conjunto das análises qualitativa e quantitativa apontou para o uso preferencial do meio Amies para o transporte de suabes nasais. O segundo experimento indicou que o ágar MacConkey e o G20G modificado mostraram resultados similares e foram superiores ao meio de ceftiofur. No experimento final, a técnica de reação em cadeia pela polimerase apresentou uma capacidade de detecção da B. bronchiseptica superior a do cultivo.

Three comparative assays were performed seeking to improve the sensitivity of the diagnosis of Bordetella bronchiseptica infection analyzing swine nasal swabs. An initial assay compared the recovery of B. bronchiseptica from swabs simultaneously inoculated with B. bronchiseptica and some interfering bacteria, immersed into three transport formulations (Amies with charcoal, trypticase soy broth and phosphate buffer according to Soerensen supplemented with 5% of bovine fetal serum) and submitted to different temperatures (10ºC and 27ºC) and periods of incubation (24, 72 and 120 hours). A subsequent assay compared three selective media (MacConkey agar, modified selective medium G20G and a ceftiofur medium) for their recovery capabilities from clinical specimens. One last assay compared the polymerase chain reaction to the three selective media. In the first assay, the recovery of B. bronchiseptica from transport systems was better at 27ºC and the three formulations had good performances at this temperature, but the collection of qualitative and quantitative analysis indicated the advantage of Amies medium for nasal swabs transportation. The second assay indicated that MacConkey agar and modified G20G had similar results and were superior to the ceftiofur medium. In the final assay, polymerase chain reaction presented superior capability of B. bronchiseptica detection to culture procedures.

Clostridium perfringens é um importante agente infeccioso em medicina veterinária e humana. Em suínos, o agente é responsável pela enterite necrótica e enterotoxemia, caracterizadas por diarréia, perda de peso, atraso no desenvolvimento e morte. No presente estudo foi utilizado o polimorfismo do comprimento de fragmentos amplificados (AFLP), para caracterizar 54 isolados de C. perfringens obtidos de suínos com diarréia. A análise dos resultados do AFLP demonstrou 29 perfis distintos com índice discriminatório igual a 0,97. A correlação entre a origem dos isolados e os agrupamentos obtidos foi parcial, sendo apenas possível a correlação total de 18,5% das amostras estudadas. A caracterização das cepas em biotipos (A, B, C, D e E), produção da toxina beta-2 e enterotoxina foi realizada através da reação da polimerase em cadeia (PCR). Dentre as cepas analisadas foram observados os biotipos A, C e D, sendo que todas as amostras foram positivas para a presença do gene codificador da toxina beta-2 e negativas para o gene codificador da enterotoxina. Neste estudo, o AFLP demonstrou ser uma metodologia simples, rápida, de baixo custo, com alto poder discriminatório e boa reprodutibilidade, apresentando grande potencial para estudos epidemiológicos envolvendo cepas de C. perfringens de origem animal.

Clostridium perfringens is an important pathogen in human and veterinary medicine. In swine, the agent is responsible for necrotic enteritis and enterotoxemia characterized by diarrhea, weight loss, delayed development and, in some cases, death. In the present study amplified fragment length polymorphism analyses (AFLP) was used to characterize 54 C. perfringens strains isolated from swine presenting diarrhea. Analysis of the results showed 29 distinct profiles with discriminatory index equal to 0.97. Partial correlation between the origin of the isolates and groups was drawn, and correlation was possible in only 18.5% of the samples. Characterization of the strains in biotypes (A, B, C, D and E), production of beta-2 toxin and enterotoxin were performed by means of the polymerase chain reaction (PCR). Biotypes A, C and D were observed among the strains analyzed. All samples were positive for presence of the gene encoding beta-2 toxin and negative for the gene encoding enterotoxin. AFLP have shown to be a simple, fast, low cost method with high discriminative power and good reproducibility, presenting a great potential in epidemiological studies involving C. perfringens strains of animal origin.

A distribuição dos genes de virulência sefC, pefA e spvC foi investigada em 110 amostras de Salmonella Enteritidis pertencentes ao fagotipo 4 através da reação em cadeia da polimerase. A influência destes genes na colonização do ceco e invasão do fígado e baço em pintinhos de um dia de idade foi avaliada. Oito amostras foram negativas para o gene spvC, três para o gene pefA e uma amostra para o gene sefC. Estes resultados permitiram a classificação das amostras em quatro genótipos. A presença destes genes não influenciou a invasão da bactéria no fígado e baço das aves dez dias após a infecção, entretanto, a presença de mais de um gene fimbrial pode ter relação com a colonização cecal.

The distribution of virulence genes, sefC, pefA and spvC, was investigated in 110 Salmonella Enteritidis phage type 4 strains by polymerase chain reaction. Their influence in the caecal colonization and invasion of liver and spleen of one-day-old chickens was studied. Eight isolates were negative for the spvC gene, three for the pefA gene and one, for the sefC gene. These results allowed grouping the strains into four genotypes. Presence of these genes did not influence bacteria invasion in the liver and spleen of the chickens ten days after infection, although the presence of more than one fimbrial gene can be related to caecal colonization.

A enterite proliferativa suína, a espiroquetose colônica, a disenteria suína e a salmonelose são as doenças entéricas mais freqüentes em suínos das fases de crescimento e terminação no Brasil. O diagnóstico destas doenças através de técnicas bacteriológicas tradicionais é difícil, lento e no caso da enterite proliferativa suína, impossível. A detecção destes agentes através da reação de polimerase em cadeia é altamente sensível e específica e está se tornando uma ferramenta muito útil no diagnóstico em Medicina Veterinária. No presente estudo foram testadas duas técnicas de PCR sob a forma de "multiplex"para detecção e identificação simultânea dos agentes bacterianos envolvidos na enterite proliferativa suína, na espiroquetose colônica, na disenteria suína e na salmonelose em amostras de fezes diarreicas. O DNA obtido de culturas puras de cada agente sozinho ou misturado em diferentes combinações e concentrações foram amplificados utilizando a Multiplex PCR específica para Lawsonia intracellularis e Salmonella, ou Brachispyra pilosicoli e Brachispyra hyodysenteriae. Após a padronização, a M-PCR foi aplicada na detecção dos quatro agentes em 541 amostras de fezes diarreicas de suínos provenientes de diferentes Estados do Brasil. O agente mais freqüente foi Lawsonia intracellularis (13%), seguido pela Salmonella spp. (4,8%), B. hyodysenteriae (1,4%), B. pilosicoli (1%) e suas diferentes associações. Os resultados obtidos indicam que as duas reações testadas permitem a detecção destes quatro importantes patógenos entéricos de maneira rápida e específica.

In Brazil, the most common bacterial enteric diseases affecting growing and finishing pigs are porcine proliferative enteritis, porcine intestinal spirochetosis, swine dysentery, and salmonellosis. The diagnosis of these diseases by routine culture techniques is expensive, difficult, time-consuming, and even impossible, in cases of porcine proliferative enteritis. The detection of pathogens by polymerase chain reaction is a highly sensitive and specific method that can be an useful tool in veterinary diagnosis. Two multiplex PCR (M-PCR) assays were tested for simultaneous detection and identification of bacterial agents associated with porcine proliferative enteritis, porcine intestinal spirochetosis, swine dysentery, and salmonellosis in diarrheic fecal samples. The DNA obtained from pure cultures of each bacterial agent or mixed in different combinations and concentrations was amplified by using Lawsonia intracellularis and Salmonella, or Brachyspira pilosicoli and Brachyspira hyodysenteriae specific M-PCR assays. After electrophoresis in agarose gel and staining, the amplification products indicated the presence of individual or simultaneous amplification of L. intracellularis and Salmonella or B. pilosicoli and B. hyodysenteriae specific DNA sequences. After standardization, the M-PCR tests were used to test 541 swine diarrheic fecal samples obtained from different regions in Brazil. The most frequently detected pathogen was Lawsonia intracellularis (13%), followed by Salmonella (4.8%), B. hyodysenteriae (1.4%), B. pilosicoli (1%) and their various associations. Results from this study suggest that the two M-PCR assays can be used for specific detection and identification of four important enteric bacterial pathogens alone or in combination.