Diagnosis of blackleg from cattle tissue impregnated in common filter paper / Diagnóstico de carbúnculo sintomático a partir de papel filtro comum impregnado com tecido bovino

Blackleg, an acute myonecrosis caused by Clostridium chauvoei, is usually underdiagnosed since the rapid transport of adequate samples for laboratory testing is difficult. This study tested a direct polymerase chain reaction (PCR) technique using common filter paper impregnated with cattle tissue samples obtained from animals suspected with blackleg. Twenty-five samples, belonging to eleven animals from Rio Grande do Sul State, Brazil, were analyzed. The direct PCR technique identified eight positive animals corroborating with results from microbiological culture. Skeletal muscle was the most common tissue type used in this study and when the animal was positive the pathogen was always detected in this tissue. Storage time of the impregnated filter paper at room temperature did not prove to be a limiting factor for the quality of the results indicating that this procedure can be carried out in the field and samples be sent in regular mail. Our results suggested that direct PCR of common filter paper impregnated with cattle tissue is a practical and economical alternative for the diagnosis of blackleg.(AU)

Carbúnculo sintomático, uma mionecrose aguda causada por Clostridium chauvoei, costuma ser subdiagnosticada, pois o transporte rápido de amostras adequadas para exames laboratoriais é complicado. O objetivo deste estudo foi testar a técnica de reação em cadeia da polimerase (PCR) direta, utilizando papel filtro comum impregnado com amostras de tecido bovino obtidas de animais suspeitos de carbúnculo sintomático. Foram analisadas 25 amostras, pertencentes a onze animais do estado do Rio Grande do Sul, Brasil. A técnica de PCR direta identificou oito animais positivos, corroborando com os resultados da cultura microbiológica. O músculo esquelético foi o tecido mais utilizado neste estudo e quando o animal foi positivo, o patógeno sempre foi detectado neste tecido. O tempo de armazenamento do papel filtro impregnado, à temperatura ambiente, não se mostrou um fator limitante para a qualidade dos resultados, indicando que esse procedimento pode ser realizado no local e as amostras enviadas por correio normal. Nossos resultados sugerem que a PCR direta usando papel filtro comum impregnado com tecido bovino é uma alternativa prática e econômica para o diagnóstico de carbúnculo sintomático.(AU)

Diagnosis of blackleg from cattle tissue impregnated in common filter paper / Diagnóstico de carbúnculo sintomático a partir de papel filtro comum impregnado com tecido bovino

Blackleg, an acute myonecrosis caused by Clostridium chauvoei, is usually underdiagnosed since the rapid transport of adequate samples for laboratory testing is difficult. This study tested a direct polymerase chain reaction (PCR) technique using common filter paper impregnated with cattle tissue samples obtained from animals suspected with blackleg. Twenty-five samples, belonging to eleven animals from Rio Grande do Sul State, Brazil, were analyzed. The direct PCR technique identified eight positive animals corroborating with results from microbiological culture. Skeletal muscle was the most common tissue type used in this study and when the animal was positive the pathogen was always detected in this tissue. Storage time of the impregnated filter paper at room temperature did not prove to be a limiting factor for the quality of the results indicating that this procedure can be carried out in the field and samples be sent in regular mail. Our results suggested that direct PCR of common filter paper impregnated with cattle tissue is a practical and economical alternative for the diagnosis of blackleg.

Carbúnculo sintomático, uma mionecrose aguda causada por Clostridium chauvoei, costuma ser subdiagnosticada, pois o transporte rápido de amostras adequadas para exames laboratoriais é complicado. O objetivo deste estudo foi testar a técnica de reação em cadeia da polimerase (PCR) direta, utilizando papel filtro comum impregnado com amostras de tecido bovino obtidas de animais suspeitos de carbúnculo sintomático. Foram analisadas 25 amostras, pertencentes a onze animais do estado do Rio Grande do Sul, Brasil. A técnica de PCR direta identificou oito animais positivos, corroborando com os resultados da cultura microbiológica. O músculo esquelético foi o tecido mais utilizado neste estudo e quando o animal foi positivo, o patógeno sempre foi detectado neste tecido. O tempo de armazenamento do papel filtro impregnado, à temperatura ambiente, não se mostrou um fator limitante para a qualidade dos resultados, indicando que esse procedimento pode ser realizado no local e as amostras enviadas por correio normal. Nossos resultados sugerem que a PCR direta usando papel filtro comum impregnado com tecido bovino é uma alternativa prática e econômica para o diagnóstico de carbúnculo sintomático.

The antibacterial and physiological effects of pure and nanoencapsulated Origanum majorana essential oil on fish infected with Aeromonas hydrophila.

This study evaluated the antibacterial activity of Origanum majorana essential oil (EOM) and nanocapsules of this oil (NOM) in silver catfish, Rhamdia quelen, infected with Aeromonas hydrophila, and addressed their effects on silver catfish hematological and metabolic parameters. Fish were inoculated with A. hydrophila (360 µL, at a concentration of 1.5 × 109 CFU mL-1) and submitted to 1 h daily baths with EOM (0 (control), 20 or 30 µL L-1), NOM (0 (control), 5 or 10 µL L-1) or a positive control containing florfenicol (30 µL L-1) called group Maxflor® for five consecutive days. All treatments improved the survival rate of the infected fish, but we suggest the treatment of A. hydrophila infections through daily baths with 20 µL L-1 EOM or 5 µL L-1 NOM for five consecutive days as these were the lowest effective concentrations tested. Silver catfish treated with EOM and NOM had higher lymphocyte levels, indicating stimulation of the immune system in these fish. The lowest liver glucose level was found in the group treated with the lowest concentration of NOM, and the lactate values in the liver and muscle of all groups were within the normal values reported for this species. In addition, nanocapsules required much less EOM to elicit effective antibacterial treatment.

Virulence factors and antimicrobial susceptibility profile of extraintestinal Escherichia coli isolated from an avian colisepticemia outbreak.

In this study an avian colisepticemia outbreak was investigated. Two isolates from a chicken with colisepticemia were characterized for antimicrobial susceptibility and virulence factors profile. For this purpose 7 antimicrobial and 29 genes (fimH, hrlA/hek, iha, papC, sfa/focCD, tsh, mat, tia, gimB, ibeA, chuA, fyuA, ireA, iroN, irp2, iucD, sitD. chr., sitD. ep., iss, neuC, ompA, traT, astA, hlyA, sat, vat, pic, malX, cvi/cva) were tested. The outbreak happened in a hick chicken breeding located in the northwestern region of Rio Grande do Sul state in South of Brazil and caused 28.3% (102 deads of a total of 360 chickens) of mortality rate. Escherichia coli isolates obtained from the avian spleen and liver belong to the same phylogenetic group A and present resistance to all antimicrobials tested (ampicillin, tetracycline, gentamicin, neomycin, sulfa + trimethoprim, enrofloxacin, and norfloxacin). Both isolates harbor virulence factors related to adhesion (fimH, papC, mat), invasion (tia), iron acquisition system (iroN) and serum resistance (iss, ompA, traT), showing that these groups are important for Avian Pathogenic E. coli (APEC). However, they present different virulence profiles for some genes, whereas liver-isolate carries more hrlA/hek (adhesin), gimB (invasin), sitD ep. (iron acquisition system), sat (toxin) and hylA (toxin) genes, the spleen-isolate harbors fyuA (iron acquisition system) gene. Here, we highlight a coinfection by different strains of APEC in the same animal with colisepticemia, the great antimicrobial resistance of these bacterial isolates and the genetic traits that modulate the virulence for high mortality rate of chickens for human consumption.

Molecular identification of Clostridium chauvoei from common filter paper / Molecular identification of Clostridium chauvoei from common filter paper

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary fi lter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA.Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common fi lter paper was tested for specifi city, sensitivity and feasibility. To test the specifi city, the papers were impregnated with a suspension of the following microorganisms: C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common fi lter paper. To both test, DNA extraction of impregnated ordinary fi lter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common fi lter paper with suspension of C. cha

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary fi lter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA.Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common fi lter paper was tested for specifi city, sensitivity and feasibility. To test the specifi city, the papers were impregnated with a suspension of the following microorganisms: C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common fi lter paper. To both test, DNA extraction of impregnated ordinary fi lter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common fi lter paper with suspension of C. cha

Molecular identification of Clostridium chauvoei from common filter paper

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary filter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA. Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common filter paper was tested for specificity, sensitivity and feasibility. To test the specificity, the papers were impregnated with a suspension of the following microorganisms: C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common filter paper. To both test, DNA extraction of impregnated ordinary filter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common filter paper with suspension of C. chauvoei. The filter paper was stored for 48 h, 72 h and one week. Subsequently, a rapid and direct PCR approach to detect C. chauvoei was performed. All procedures were performed in triplicate and was performed by PCR using the same primers employed to amplify the flic gene encoding flagellin (FliC). There was no cross reaction with any tested microorganism, confirming the specificity of the flic gene previously studied. It was possible to visualize the amplification until the corresponding to 100 CFU. Specific PCR amplification products were visualized in 100% of the trials at 48 h, 70% at 72 h, and 90% within one week of storage at room temperature using direct PCR. Discussion: This report describes a rapid, highly sensitive method for the detection of C. chauvoei DNA from liver tissue bovine samples stored on filter papers. It was observed a high sensitivity and a specificity of 100%. The selection of hepatic tissue was based on previous studies that identified C. chauvoei in this tissue by PCR assays. Besides, blackleg in visceral form can be detected in hepatic tissue but does not in muscle. According to others researchers, the direct PCR procedure exhibits several advantages, such as costs and time reduction through omission of DNA extraction as well as avoid any cross contamination with other agents. However, current substances in the blood and tissues may inhibit the PCR amplification. For this reason, a methanol fixation and preheating the samples before the direct PCR assay was performed, mainly because the amplicon is relatively large (535 bp). Some authors consider the use of direct PCR from filter paper simple and inexpensive which offer a handy tool for epidemiologic studies and to clinicians, particularly in many tropical countries where collection and storage of clinical specimens for this purpose are logistically complicated. Furthermore, this procedure can simplify the material shipment for laboratory diagnosis, since it can also be transported in standard envelops by regular mail. The current results propose the use of the direct PCR from common filter paper as practical and economical alternative to diagnosis of blackleg.

Antibacterial activity of essential oil of cinnamon and trans-cinnamaldehyde against Staphylococcus spp. isolated from clinical mastitis of cattle and goats

Background: Milk production plays a role as a source of protein and constitutes an important socioeconomic factor for small farms. The lack of planning of the creators, together with the lack of specialized technical assistance, and the precarious hygienic and sanitary management, are barriers that hinder the development of the activity. Bovine and caprine mastitis is an inflammation process of the mammary gland caused by microorganisms. Staphylococcus spp. is the main causative agent of mastitis, with higher prevalence in cases of clinical and subclinical manifestations. The most common treatment is based on intramammary infusion of antibacterial agents. However, cure rates obtained with such drugs are not always effectives, because it may determine the emergence of resistant bacteria as well increase amounts of antibiotic residues in milk. Nevertheless, the treatment of bovine subclinical mastitis caused by Staphylococcus spp. in the lactation can be economically unviable. Alternative treatments of bovine mastitis with plant derived compounds have been described. The investigation of its antimicrobial activity against bacterial agents of mastitis is justifiable. Materials, Methods & Results: This study evaluated the antimicrobial activity of essential oil (EO) of Cinnamomum zeylanicum (cinnamon) and the fraction trans-cinnamaldehyde (TC) against 65 isolates of Staphylococcus spp. from mastitis of cows (n = 32) and goats (n = 33). The cinnamon EO was extracted from the leaves and was composed majority by isoeugenol (96%), while the TC was acquired commercially. The antimicrobial susceptibility tests were performed according to CLSI M31-A3 agar diffusion method. Based on these tests, the isolates were divided into subgroups according to the resistance profiles. Subsequently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by broth microdilution technique CLSI M31-A3. The MICs (geometric mean, GM) for cinnamon EO were 2032 mg/L and 2102 mg/L, respectively, for cattle and goats, while the majority fraction TC was most active with MICs (GM) of 598 mg/L and 875 mg/L, respectively, for cattle and goats. Discussion: Some differences between these results and those reported by other authors can be attributed to the composition of OEs or techniques employed. In this study, we used the broth microdilution technique based on the document M31-A3, from CLSI, for the determination of MICs and MBCs. The lack of an internationally standardized technique for evaluating OEs and plant extracts that allow different protocols undermines the comparisons of results. The results of this study confirmed the antimicrobial activity of cinnamon EO and TC against clinical Staphylococcus spp. independent of the antibacterial susceptibility profile. The best antimicrobial activity was observed with TC than in the cinnamon EO, which deserves further research investigating the use of these compound in the treatment of mastitis. The TC can be found in high concentration in cinnamon bark. However, the essential oil used in this work was extracted from the leaves, and the major component was isoeugenol. The cinnamon EO composition can be varied depending on the part of the plant where it is extracted.

Conjuntivite bacteriana secundária à doença dentária em chinchilas (Chinchilla lanigera)

Three cases of bacterial conjunctivitis associated with severe dental disease in chinchillas are described. All affected chinchillas had a history of weight loss, ptyalism, and epiphora which progressed to suppurative ocular exsudation. At necropsy incisor and molar teeth revealed marked elongation of the clinical crown and overgrowth of the dental apexes resulting in deformation of the tear ducts. Histologically, there was neutrophilic infiltrate in the conjunctiva and eyelid skin. Microbiological culture carried out in samples from the ocular exsudate yielded Staphylococcus coagulase-positive. Compression of the lacrimal duct by dental overgrowth compromised tear draining and resulting in epiphora. This is a major predisposing factor inducing bacterial ocular infection in chinchillas.

São relatados três casos de conjuntivite bacteriana associada à doença dentária grave em chinchilas. Todas as chinchilas afetadas tinham histórico de emagrecimento, hipersalivação e epífora, que progrediu para exsudação ocular purulenta. Durante a necropsia, foi constatado marcado alongamento da coroa clínica dos incisivos e molares, e crescimento do ápice dentário, causando deformação óssea e compressão do canal lacrimal. Histologicamente, observou-se infiltrado de neutrófilos na conjuntiva e pálpebras. Cultivo microbiológico do exsudato ocular revelou crescimento de Staphylococcus coagulase-positiva . Uma compressão do canal lacrimal pelo crescimento dentário excessivo impediu a drenagem das lágrimas, resultando em epífora. Esse é um importante fator predisponente para infecção bacteriana ocular em chinchilas.

Conjuntivite bacteriana secundária à doença dentária em chinchilas (Chinchilla lanigera)

Three cases of bacterial conjunctivitis associated with severe dental disease in chinchillas are described. All affected chinchillas had a history of weight loss, ptyalism, and epiphora which progressed to suppurative ocular exsudation. At necropsy incisor and molar teeth revealed marked elongation of the clinical crown and overgrowth of the dental apexes resulting in deformation of the tear ducts. Histologically, there was neutrophilic infiltrate in the conjunctiva and eyelid skin. Microbiological culture carried out in samples from the ocular exsudate yielded Staphylococcus coagulase-positive. Compression of the lacrimal duct by dental overgrowth compromised tear draining and resulting in epiphora. This is a major predisposing factor inducing bacterial ocular infection in chinchillas.

São relatados três casos de conjuntivite bacteriana associada à doença dentária grave em chinchilas. Todas as chinchilas afetadas tinham histórico de emagrecimento, hipersalivação e epífora, que progrediu para exsudação ocular purulenta. Durante a necropsia, foi constatado marcado alongamento da coroa clínica dos incisivos e molares, e crescimento do ápice dentário, causando deformação óssea e compressão do canal lacrimal. Histologicamente, observou-se infiltrado de neutrófilos na conjuntiva e pálpebras. Cultivo microbiológico do exsudato ocular revelou crescimento de Staphylococcus coagulase-positiva . Uma compressão do canal lacrimal pelo crescimento dentário excessivo impediu a drenagem das lágrimas, resultando em epífora. Esse é um importante fator predisponente para infecção bacteriana ocular em chinchilas.

Molecular identification of Clostridium chauvoei from common filter paper / Molecular identification of Clostridium chauvoei from common filter paper

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary fi lter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA.Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common fi lter paper was tested for specifi city, sensitivity and feasibility. To test the specifi city, the papers were impregnated with a suspension of the following microorganisms: C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common fi lter paper. To both test, DNA extraction of impregnated ordinary fi lter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common fi lter paper with suspension of C. cha

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary fi lter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA.Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common fi lter paper was tested for specifi city, sensitivity and feasibility. To test the specifi city, the papers were impregnated with a suspension of the following microorganisms: C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common fi lter paper. To both test, DNA extraction of impregnated ordinary fi lter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common fi lter paper with suspension of C. cha

Surto de listeriose sistêmica em chinchilas

Listeriosis is an infectious disease, which affects a variety of animal species and cause septicemia, encephalitis and abortion. Chinchillas are the most susceptible animals to the systemic infection by Listeria monocytogenes. This report describes an outbreak of systemic listeriosis in a farm of chinchillas in the Central region of the Rio Grande do Sul state, Brazil. Mortality rate was about 16%. On necropsy, there were multiple random small white foci on the liver capsule and parenchyma and enlargement of the hepatic lymph node. Histologically, there were multifocal necro-suppurative hepatitis and suppurative lymphadenitis with numerous intralesional bacilli. L. monocytogenes was the etiology of the systemic listeriosis outbreak. The diagnosis was based on gross and microscopic lesions, genotypical and phenotypical characteristics and by immunohistochemistry technique.

A listeriose é uma doença infecciosa que afeta uma grande variedade de espécies animais, causando septicemia, encefalite e aborto. As chinchilas são os animais mais susceptíveis à infecção sistêmica por Listeria monocytogenes. Este relato descreve um surto de listeriose sistêmica em uma criação de chinchilas da região central do Estado do Rio Grande do Sul, onde cerca de 16% das chinchilas morreram. Na necropsia, havia múltiplos focos brancos, pequenos e de distribuição aleatória nas superfícies capsular e de corte do fígado e aumento de volume do linfonodo hepático. Histologicamente, observaram-se hepatite necrossupurativa e linfadenite supurativa multifocais, com numerosos bacilos intralesionais. L. monocytogenes foi o agente etiológico do surto de listeriose sistêmica, sendo o diagnóstico confirmado por meio das lesões de necropsia e histopatológicas, das características fenotípicas e genotípicas da bactéria e da técnica de imuno-histoquímica.

Surto de listeriose sistêmica em chinchilas

Listeriosis is an infectious disease, which affects a variety of animal species and cause septicemia, encephalitis and abortion. Chinchillas are the most susceptible animals to the systemic infection by Listeria monocytogenes. This report describes an outbreak of systemic listeriosis in a farm of chinchillas in the Central region of the Rio Grande do Sul state, Brazil. Mortality rate was about 16%. On necropsy, there were multiple random small white foci on the liver capsule and parenchyma and enlargement of the hepatic lymph node. Histologically, there were multifocal necro-suppurative hepatitis and suppurative lymphadenitis with numerous intralesional bacilli. L. monocytogenes was the etiology of the systemic listeriosis outbreak. The diagnosis was based on gross and microscopic lesions, genotypical and phenotypical characteristics and by immunohistochemistry technique.

A listeriose é uma doença infecciosa que afeta uma grande variedade de espécies animais, causando septicemia, encefalite e aborto. As chinchilas são os animais mais susceptíveis à infecção sistêmica por Listeria monocytogenes. Este relato descreve um surto de listeriose sistêmica em uma criação de chinchilas da região central do Estado do Rio Grande do Sul, onde cerca de 16% das chinchilas morreram. Na necropsia, havia múltiplos focos brancos, pequenos e de distribuição aleatória nas superfícies capsular e de corte do fígado e aumento de volume do linfonodo hepático. Histologicamente, observaram-se hepatite necrossupurativa e linfadenite supurativa multifocais, com numerosos bacilos intralesionais. L. monocytogenes foi o agente etiológico do surto de listeriose sistêmica, sendo o diagnóstico confirmado por meio das lesões de necropsia e histopatológicas, das características fenotípicas e genotípicas da bactéria e da técnica de imuno-histoquímica.

Immunogenicity and cross reactivity indices of Streptococcus equi subsp. equi strains isolated from cases of Strangles and commercial vaccines

Horse Strangles appears frequently in animals vaccinated with commercial bacterins in Rio Grande do Sul, Brazil. Aiming to know the antigenic relationships of strains recovered from sick animals among them and with two vaccines profusely used in the state, bilateral cross reactivity indices (CRI) were estimated. In addition, the immunogenicity of vaccines prepared with field isolates and commercial vaccines was tested in mice. Antibody titers were measured by ELISA and expressed as seroconversions. Thirteen strains of Streptococcus equi subsp. equi, nine classified biochemically as typical and other four as atypical strains, were recovered from 35 sick horses belonging to 10 herds of Rio Grande do Sul, Brazil. The strains recovered from sick horses showed very close CRI, suggesting antigenic homogeneity among them, but not with the vaccinal strains. A vaccine produced with an atypical strain induced the highest seroconversion, 9.4, while two produced with typical strains were poorly-immunogenic. The commercial vaccines were less immunogenic than five and four vaccines produced with field strains, inducing seroconversions of 2.6 and 3.8, respectively.

A Adenite Eqüina é freqüente em animais vacinados com bacterinas comerciais no Rio Grande do Sul. Com o objetivo de determinar as relações antigênicas entre cepas isoladas de casos clínicos e duas vacinas amplamente utilizadas no Estado, foram determinados os índices de reatividade cruzada bilateral (IRC) entre elas. Também foi determinada a imunogenicidade em camundongos de vacinas preparadas com cepas de campo e de vacinas comerciais. Os títulos de anticorpos foram determinados por ELISA e expressos como soroconversão. Foram isoladas 13 cepas de Streptococcus equi subsp. equi de 35 casos clínicos de Adenite Eqüina em 10 diferentes rebanhos do Rio Grande do Sul, Brasil, das quais nove foram classificadas bioquimicamente como típicas e outras quatro como atípicas. As cepas de campo apresentaram elevada reatividade cruzada, sugerindo homogeneidade antigênica entre elas, mas não com as cepas vacinais. Uma vacina produzida com cepa atípica induziu soroconversão de 9.4, enquanto duas vacinas produzidas com cepas típicas foram pouco imunogênicas. As vacinas comerciais induziram soroconversões de 2.6 e 3.8, menores que quatro e cinco vacinas produzidas com cepas de campo, respectivamente.

Adenite equina: sua etiologia, diagnóstico e controle

Strangles is a contagious disease of the respiratory tract of horses produced by Streptococcus equi subsp. equi, a Lancefield's group C haemolytic bacterium. It produces a mucopurulent secretion of the anterior airways, as well as lymphadenitis and abscesses. The bacteria synthesizes several pathogenicity factors such a hyaluronic acid capsule, hyaluronidase, streptolisin O, streptokinase, IgG Fc receptors, peptidoglican and protein M. Among these factors, protein M deserves special importance due to its antifagocitic and adherence properties. The disease has high morbidity and low lethality, and produces economic losses due to low performance and treatment. Clinical diagnosis and treatment are done easily, but prophylaxis is hampered by the low potency of vaccines, that protect around 50 % of vaccinated animals. Strangles may occur during all the year, but cold and humid weather favors the survival of streptococci, making animals that live in regions with those characteristics more prone to infection. New vaccines using purified or subunit antigens have been developed aiming to increase their potency and to avoid undesirable side effects. The demonstration that bacteria strains show differences in their antigenicity, called attention on the selection of appropriate strains to use as antigens.

A adenite equina, também conhecida como garrotilho, é uma enfermidade bacteriana contagiosa, causada por Streptococcus equi, subsp. equi, bactéria hemolítica do grupo C de Lancefield, que afeta o trato respiratório anterior de equinos de todas as idades, com maior prevalência entre um e cinco anos de idade. Caracteriza-se por produzir secreção mucopurulenta das vias aéreas anteriores e linfadenite dos gânglios retrofaríngeos e submandibulares com formação de abscessos. Fatores de virulência de S. equi, subsp. equi, incluem cápsula de ácido hialurônico, hialuronidase, estreptolisina O, estreptoquinase, receptores para Fc de IgG, peptidoglicano e proteína M. Dentre esses fatores, a proteína M tem especial importância por ser de membrana com propriedades antifagocitárias e de aderência. A doença tem baixa letalidade e alta morbidade e seus prejuízos econômicos devem-se à perda de performance e custo do tratamento. O diagnóstico clínico e o tratamento não apresentam dificuldades, mas a profilaxia é prejudicada pela baixa eficiência das vacinas disponíveis, com índices de proteção de 50%. O garrotilho pode ocorrer em todas as épocas do ano, mas o frio e a umidade facilitam a sobrevivência do agente e sua disseminação, portanto animais que vivem nos estados mais frios e úmidos do país são mais vulneráveis à infecção. Novas vacinas utilizando antígenos purificados ou de subunidades estão sendo desenvolvidas com a finalidade de incrementar sua potência e evitar efeitos indesejáveis. A comprovação de diferenças de antigenicidade, entre estirpes, alerta sobre a importância da seleção apropriada das cepas vacinais.

Immunogenicity and cross reactivity indices of Streptococcus equi subsp. equi strains isolated from cases of Strangles and commercial vaccines

Horse Strangles appears frequently in animals vaccinated with commercial bacterins in Rio Grande do Sul, Brazil. Aiming to know the antigenic relationships of strains recovered from sick animals among them and with two vaccines profusely used in the state, bilateral cross reactivity indices (CRI) were estimated. In addition, the immunogenicity of vaccines prepared with field isolates and commercial vaccines was tested in mice. Antibody titers were measured by ELISA and expressed as seroconversions. Thirteen strains of Streptococcus equi subsp. equi, nine classified biochemically as typical and other four as atypical strains, were recovered from 35 sick horses belonging to 10 herds of Rio Grande do Sul, Brazil. The strains recovered from sick horses showed very close CRI, suggesting antigenic homogeneity among them, but not with the vaccinal strains. A vaccine produced with an atypical strain induced the highest seroconversion, 9.4, while two produced with typical strains were poorly-immunogenic. The commercial vaccines were less immunogenic than five and four vaccines produced with field strains, inducing seroconversions of 2.6 and 3.8, respectively.

A Adenite Eqüina é freqüente em animais vacinados com bacterinas comerciais no Rio Grande do Sul. Com o objetivo de determinar as relações antigênicas entre cepas isoladas de casos clínicos e duas vacinas amplamente utilizadas no Estado, foram determinados os índices de reatividade cruzada bilateral (IRC) entre elas. Também foi determinada a imunogenicidade em camundongos de vacinas preparadas com cepas de campo e de vacinas comerciais. Os títulos de anticorpos foram determinados por ELISA e expressos como soroconversão. Foram isoladas 13 cepas de Streptococcus equi subsp. equi de 35 casos clínicos de Adenite Eqüina em 10 diferentes rebanhos do Rio Grande do Sul, Brasil, das quais nove foram classificadas bioquimicamente como típicas e outras quatro como atípicas. As cepas de campo apresentaram elevada reatividade cruzada, sugerindo homogeneidade antigênica entre elas, mas não com as cepas vacinais. Uma vacina produzida com cepa atípica induziu soroconversão de 9.4, enquanto duas vacinas produzidas com cepas típicas foram pouco imunogênicas. As vacinas comerciais induziram soroconversões de 2.6 e 3.8, menores que quatro e cinco vacinas produzidas com cepas de campo, respectivamente.

Adenite equina: sua etiologia, diagnóstico e controle

Strangles is a contagious disease of the respiratory tract of horses produced by Streptococcus equi subsp. equi, a Lancefield's group C haemolytic bacterium. It produces a mucopurulent secretion of the anterior airways, as well as lymphadenitis and abscesses. The bacteria synthesizes several pathogenicity factors such a hyaluronic acid capsule, hyaluronidase, streptolisin O, streptokinase, IgG Fc receptors, peptidoglican and protein M. Among these factors, protein M deserves special importance due to its antifagocitic and adherence properties. The disease has high morbidity and low lethality, and produces economic losses due to low performance and treatment. Clinical diagnosis and treatment are done easily, but prophylaxis is hampered by the low potency of vaccines, that protect around 50 % of vaccinated animals. Strangles may occur during all the year, but cold and humid weather favors the survival of streptococci, making animals that live in regions with those characteristics more prone to infection. New vaccines using purified or subunit antigens have been developed aiming to increase their potency and to avoid undesirable side effects. The demonstration that bacteria strains show differences in their antigenicity, called attention on the selection of appropriate strains to use as antigens.

A adenite equina, também conhecida como garrotilho, é uma enfermidade bacteriana contagiosa, causada por Streptococcus equi, subsp. equi, bactéria hemolítica do grupo C de Lancefield, que afeta o trato respiratório anterior de equinos de todas as idades, com maior prevalência entre um e cinco anos de idade. Caracteriza-se por produzir secreção mucopurulenta das vias aéreas anteriores e linfadenite dos gânglios retrofaríngeos e submandibulares com formação de abscessos. Fatores de virulência de S. equi, subsp. equi, incluem cápsula de ácido hialurônico, hialuronidase, estreptolisina O, estreptoquinase, receptores para Fc de IgG, peptidoglicano e proteína M. Dentre esses fatores, a proteína M tem especial importância por ser de membrana com propriedades antifagocitárias e de aderência. A doença tem baixa letalidade e alta morbidade e seus prejuízos econômicos devem-se à perda de performance e custo do tratamento. O diagnóstico clínico e o tratamento não apresentam dificuldades, mas a profilaxia é prejudicada pela baixa eficiência das vacinas disponíveis, com índices de proteção de 50%. O garrotilho pode ocorrer em todas as épocas do ano, mas o frio e a umidade facilitam a sobrevivência do agente e sua disseminação, portanto animais que vivem nos estados mais frios e úmidos do país são mais vulneráveis à infecção. Novas vacinas utilizando antígenos purificados ou de subunidades estão sendo desenvolvidas com a finalidade de incrementar sua potência e evitar efeitos indesejáveis. A comprovação de diferenças de antigenicidade, entre estirpes, alerta sobre a importância da seleção apropriada das cepas vacinais.

Immunogenicity and cross reactivity indices of Streptococcus equi subsp. equi strains isolated from cases of Strangles and commercial vaccines

Horse Strangles appears frequently in animals vaccinated with commercial bacterins in Rio Grande do Sul, Brazil. Aiming to know the antigenic relationships of strains recovered from sick animals among them and with two vaccines profusely used in the state, bilateral cross reactivity indices (CRI) were estimated. In addition, the immunogenicity of vaccines prepared with field isolates and commercial vaccines was tested in mice. Antibody titers were measured by ELISA and expressed as seroconversions. Thirteen strains of Streptococcus equi subsp. equi, nine classified biochemically as typical and other four as atypical strains, were recovered from 35 sick horses belonging to 10 herds of Rio Grande do Sul, Brazil. The strains recovered from sick horses showed very close CRI, suggesting antigenic homogeneity among them, but not with the vaccinal strains. A vaccine produced with an atypical strain induced the highest seroconversion, 9.4, while two produced with typical strains were poorly-immunogenic. The commercial vaccines were less immunogenic than five and four vaccines produced with field strains, inducing seroconversions of 2.6 and 3.8, respectively.

A Adenite Eqüina é freqüente em animais vacinados com bacterinas comerciais no Rio Grande do Sul. Com o objetivo de determinar as relações antigênicas entre cepas isoladas de casos clínicos e duas vacinas amplamente utilizadas no Estado, foram determinados os índices de reatividade cruzada bilateral (IRC) entre elas. Também foi determinada a imunogenicidade em camundongos de vacinas preparadas com cepas de campo e de vacinas comerciais. Os títulos de anticorpos foram determinados por ELISA e expressos como soroconversão. Foram isoladas 13 cepas de Streptococcus equi subsp. equi de 35 casos clínicos de Adenite Eqüina em 10 diferentes rebanhos do Rio Grande do Sul, Brasil, das quais nove foram classificadas bioquimicamente como típicas e outras quatro como atípicas. As cepas de campo apresentaram elevada reatividade cruzada, sugerindo homogeneidade antigênica entre elas, mas não com as cepas vacinais. Uma vacina produzida com cepa atípica induziu soroconversão de 9.4, enquanto duas vacinas produzidas com cepas típicas foram pouco imunogênicas. As vacinas comerciais induziram soroconversões de 2.6 e 3.8, menores que quatro e cinco vacinas produzidas com cepas de campo, respectivamente.

Virulence factors and antimicrobial resistance of escherichia coli isolated from urinary tract of swine in southern of Brazil

The present study determined the molecular and resistance patterns of E. coli isolates from urinary tract of swine in Southern of Brazil. Molecular characterization of urinary vesicle samples was performed by PCR detection of virulence factors from ETEC, STEC and UPEC. From a total of 82 E. coli isolates, 34 (38.63%) harbored one or more virulence factors. The frequency of virulence factors genes detected by PCR were: pap (10.97%), hlyA (10.97%), iha (9.75%), lt (8.53%), sta (7.31%) sfa (6.09%), f4 (4.87%), f5 (4.87%), stb (4.87%), f6 (1.21%) and f41 (1.21%). Isolates were resistant to penicillin (95.12%), lincomycin (93.9%), erythromycin (92.68%), tetracycline (90.24%), amoxicillin (82.92%), ampicillin (74.39%), josamycin (79.26%), norfloxacin (58.53%), enrofloxacin (57.31%), gentamicin (39.02%), neomycin (37.8%), apramycin (30.48%), colistine (30.48%) and cefalexin (6.09%). A number of 32 (39.02%) E. coli isolates harbored plasmids.

O presente estudo teve por objetivo determinar os padrões moleculares e de resistência aos antimicrobianos de isolados de E. coli provenientes do trato urinário de suínos no Sul do Brasil. Os fatores estudados dividiram os patotipos ETEC, STEC e UPEC. Trinta e quatro (38,63%) isolados avaliados apresentavam um ou mais dos fatores de virulência pesquisados. A freqüência dos genes de virulência detectados foram: pap (10,97%), hlyA (10,97%), iha (9,75%), lt (8,53%), sta (7,31%) sfa (6,09%), f4 (4,87%), f5 (4,87%), stb (4,87%), f6 (1,21%) e f41 (1,21%). Os isolados foram resistentes à penicilina (95,12%), lincomicina (93,9%), eritromicina (92,68%), tetraciclina (90,24%), amoxacilina (82,92%), ampicilina (74,39%), josamicina (79,26%), norfloxacina (58,53%), enrofloxacina (57,31%), gentamicina (39,02%), neomicina (37,8%), apramicina (30,48%), colistina (30,48%) e cefalexina (6,09%). Trinta e dois (39,02%) isolados de E. coli continham plasmídeos.

Molecular characterization of Rhodococcus equi isolates of horse breeding farms from an endemic region in South of Brazil by multiplex PCR

Rhodococcus equi is a gram-positive coco-bacillus and an intracellular opportunistic pathogen which causes pneumonia in foals. It is widely detected in environment and has been isolated from several sources, as soil, feces and gut from health and sick foals. The goal of this study was to characterize the epidemiological status (endemic, sporadic or no infection) of horse breeding farms from Bage County in South of Brazil, using a multiplex PCR. One hundred and eighteen R. equi isolates were identified by biochemical tests and submitted to a specie-specific and vapA multiplex PCR. These isolates were obtained from: three farms where the R. equi infection has been noticed, two farms where the disease has been not reported and one farm where the disease is frequent. All clinical isolates from horse breeding farms where the disease is endemic and/or sporadic were vapA-positive. None environmental isolates were vapA-positive. In three horse breeding farms with sporadic R. equi infection, 11.54% of the isolates from adult horse feces were vapA-positive. The multiplex PCR technique has proven to be effective for the molecular and epidemiological characterization of the R. equi isolates in horse breeding farms. An important finding in this study was the isolation of vapA-positive R. equi from adult horse feces, which is an evidence for other routes of dissemination of this pathogen in the farms.

Rhodococcus equi é um coco-bacilo gram positivo que causa pneumonia em potros. Trata-se de um patógeno oportunista amplamente detectado no ambiente e isolado de várias fontes, como solo, fezes e intestino de potros doentes e sadios. O presente estudo visa caracterizar a situação epidemiológica de criatórios eqüinos da região de Bagé, RS, Brasil, pela técnica de PCR multiplex. Cento e dezoito isolados de R. equi foram identificados por testes bioquímicos e, posteriormente, submetidos a um PCR multiplex para caracterização da espécie e da presença do gene vapA. Estes isolados eram provenientes de três haras com histórico da doença, dois haras onde não havia casos da doença e uma propriedade onde a infecção por R. equi é relatada frequentemente. Todos os isolados clínicos provenientes de haras onde a doença é endêmica e/ou esporádica foram vapA positivos. Nenhum isolado ambiental foi vapA positivo. Nos três haras onde a doença é esporádica, 11,54% dos isolados de fezes de eqüinos adultos foram positivos para o gene vapA. A técnica de PCR multiplex mostrou-se efetiva para caracterização epidemiológica e molecular dos criatórios equinos, estando de acordo com o histórico da propriedade. Um fato relevante demonstrado pela aplicação desta técnica foi a detecção de R. equi vapA positivo nas fezes de eqüinos adultos. Esta observação pode pressupor que haja outras vias de disseminação da bactéria dentro de uma propriedade.

ISOLAMENTO E CARACTERIZAÇÃO DE Klebsiella pneumoniae EM VEADO CATINGUEIRO (Mazama spp) - RELATO DE CASO

Klebsiella pneumoniae é uma enterobactéria associada a diversas infecções nos seres humanos e animais. A patogenia ocasionada por esta bactéria é importante nos animais silvestres onde atua como um agente oportunista. Este relato descreve o isolamento de K. pneumoniae a partir de um filhote de veado catingueiro, macho, mantido confinado no município de Petrolina-PE. O animal apresentou morte súbita antecedida por sinais nervosos de pedalagem e opistótono. Após a necropsia, o cultivo bacteriano foi realizado a partir de diversos fragmentos de fígado, intestinos, baço e pulmões. Realizou-se o teste de sensibilidade aos antimicrobianos das bactérias isoladas. Também removeu-se fragmentos dos órgãos para análise histopatológica. K. pneumoniae obteve-se em cultivos puros do fígado e pulmões. Estes isolados foram sensíveis a norfloxacina, tetraciclina, colistina, estreptomicina, neomicina, gentamicina, enrofloxacina, azitromicina, sulfazotrim e resistentes a ampicilina, penicilina, rifampicina e eritromicina. A análise histopatológica revelou congestão difusa em diversos órgãos. Relatos como estes são importantes no intuito de alertar profissionais da área a respeito da etiologia e perfil de sensibilidade aos antimicrobianos de K. pneumoniae em animais silvestres.