Effects of Pasturella Multocida B:2 and its immunogens (LPS and OMP) on reproductive hormones in Nili-Ravi Buffaloes

Abstract Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P 0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.

Resumo A pecuária é uma parte fundamental da indústria agrícola no Paquistão e contribui com 11,53% do PIB nacional. Entre as espécies de gado, os búfalos são considerados o ouro negro do Paquistão. Sendo os maiores produtores de leite em todo o mundo, os búfalos Nili-Ravi são os mais famosos. Os búfalos são afetados por muitas doenças endêmicas, entre as quais a septicemia hemorrágica (SH). Este estudo busca verificar os efeitos da exposição experimental de P. multocida B:2 (oral) e seus imunógenos, ou seja, LPS (oral e intravenoso) e OMP (oral e subcutâneo), nos perfis hormonais reprodutivos em búfalos Nili-Ravi. Amostras de soro repetidas foram coletadas da veia jugular de animais experimentais por 21 dias (0, 2, 4, 8, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 e 504 horas). Os ensaios hormonais para determinar as concentrações séricas do hormônio liberador de gonadotrofina (GnRH), hormônio foliculoestimulante (FSH), hormônio luteinizante (LH), estrogênio (E2) e progesterona (P4) foram realizados usando kits comerciais Elisa (MyBioSource). O perfil hormonal de todos os grupos de tratamento das novilhas bubalinas apresentou variações significativas (P 0,05) em relação ao grupo controle (G-1). Esses resultados indicam supressão no perfil hormonal reprodutivo de búfalos Nili-Ravi na exposição a P. multocida B:2 e seus imunógenos. Essa influência garante que a exposição à SH possa ser uma possível razão para o atraso da puberdade e o baixo desempenho reprodutivo em búfalos Nili-Ravi.

Effects of Pasturella Multocida B: 2 and its immunogens (LPS and OMP) on reproductive hormones in Nili-Ravi Buffaloes / Efeitos de Pasteurella multocida B: 2 e seus imunógenos (LPS e OMP) em hormônios reprodutivos em búfalos Nili-Ravi

Abstract Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P<0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.

Resumo A pecuária é uma parte fundamental da indústria agrícola no Paquistão e contribui com 11,53% do PIB nacional. Entre as espécies de gado, os búfalos são considerados o ouro negro do Paquistão. Sendo os maiores produtores de leite em todo o mundo, os búfalos Nili-Ravi são os mais famosos. Os búfalos são afetados por muitas doenças endêmicas, entre as quais a "septicemia hemorrágica" (SH). Este estudo busca verificar os efeitos da exposição experimental de P. multocida B:2 (oral) e seus imunógenos, ou seja, LPS (oral e intravenoso) e OMP (oral e subcutâneo), nos perfis hormonais reprodutivos em búfalos Nili-Ravi. Amostras de soro repetidas foram coletadas da veia jugular de animais experimentais por 21 dias (0, 2, 4, 8, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 e 504 horas). Os ensaios hormonais para determinar as concentrações séricas do hormônio liberador de gonadotrofina (GnRH), hormônio foliculoestimulante (FSH), hormônio luteinizante (LH), estrogênio (E2) e progesterona (P4) foram realizados usando kits comerciais Elisa (MyBioSource). O perfil hormonal de todos os grupos de tratamento das novilhas bubalinas apresentou variações significativas (P < 0,05) em relação ao grupo controle (G-1). Esses resultados indicam supressão no perfil hormonal reprodutivo de búfalos Nili-Ravi na exposição a P. multocida B:2 e seus imunógenos. Essa influência garante que a exposição à SH possa ser uma possível razão para o atraso da puberdade e o baixo desempenho reprodutivo em búfalos Nili-Ravi.

Neumonía por Pasteurella multocida y Staphylococcus aureus en una paciente sin patología subyacente / Pneumonia by Pasteurella multocida and Staphylococcus aureus in a patient whithout underlying conditions / Pneumonia por Pasteurella multocida e Staphylococcus aureus em uma paciente sem patologia subjacente

Resumen Muchos microorganismos capaces de producir infecciones en el ser humano son de origen zoonótico. Entre éstos está Pasteurella multocida, la cual se encuentra como comensal y/o patógeno oportunista, colonizando el tracto gastrointestinal y respiratorio de gran cantidad de mamíferos y aves. Por otro lado, Staphylococcus aureus es un colonizante de piel y nasofaringe y un patógeno oportunista para seres humanos y animales y su transmisión podría ser bidireccional. Se presenta un caso de neumonía por P. multocida y S. aureus en una mujer sin enfermedad subyacente, que acogía perros y gatos abandonados e ingresó con politraumatismo por caída desde un quinto piso. Ambos microorganismos fueron recuperados en una muestra de aspirado traqueal e identificados por Vitek®2 Compact. La identificación de P. multocida fue confirmada por espectrometría de masas (MALDI-TOF-MS) y presentó sensibilidad a penicilina, cefotaxima, amoxicilina-ácido clavulánico, ciprofloxacina, tetraciclina y trimetoprima-sulfametoxazol. S. aureus era sensible a meticilina sin resistencia acompañante.

Abstract Many microorganisms capable of producing infections in humans are of zoonotic origin. Among them, Pasteurella multocida, is a commensal and/or opportunistic pathogen that colonises the gastrointestinal and the respiratory tract of a great number of mammals and birds. On the other hand, Staphylococcus aureus is a coloniser of skin and nasopharynx and an opportunistic pathogen in humans and animals, whose transmission might be bidirectional. A case of pneumonia by P. multocida and S. aureus was presented in a woman without an underlying condition, who sheltered stray dogs and cats and was admitted with multiple trauma due to a fall from a fifth floor. Both microorganisms were recovered from a tracheal aspirate sample and identified by Vitek®2 Compact. Identification of P. multocida was confirmed by mass spectrometry (MALDI-TOF MS) and was susceptible to penicillin, cefotaxime, amoxicillin-clavulanic acid, ciprofloxacin, tetracycline and trimethoprim-sulfamethoxazole. S. aureus was methicillin-susceptible without accompanying resistance.

Resumo Muitos dos microrganismos capazes de produzir infecções no ser humano têm origem zoonótico. Entre eles está a Pasteurella multocida, que se encontra como comensal e/ou patógeno oportunista, colonizando o trato gastrintestinal e respiratório de grande quantidade de mamíferos e aves. Por outro lado, Staphylococcus aureus é um colonizador de pele e nasofaringe e um patógeno oportunista para seres humanos e animais e sua transmissão poderia ser bidirecional. Apresenta-se um caso de pneumonia por P. multocida e S. aureus em uma mulher, sem doença subjacente, que abrigava cães e gatos abandonados e entrou com múltiplos traumatismos devido à queda desde o quinto andar. Ambos os microrganismos foram obtidos da amostra de aspiração traqueal e identificados por Vitek®2 Compact. A identificação de P. multocida foi validada por espectrometria de massas (MALDI-TOF MS) e apresentou sensibilidade à penicilina, cefotaxima, amoxicilina-ácido clavulânico, ciprofloxacina, tetraciclina e trimetoprim-sulfometoxazol. S. aureus era meticilino sensível sem resistência acompanhante.

Microbiología de las infecciones causadas por mordeduras de perros y gatos en personas: una revisión / Microbiology of infections caused by dog and cat bites: a review

INTRODUCCIÓN: Las mordeduras causadas por animales de compañía constituyen el 5% de las heridas traumáticas registradas en los servicios de urgencias. OBJETIVO: Conocer los principales agentes infecciosos presentes en las mordeduras provocadas por perros y gatos, tanto de forma individual como conjunta; así como los factores que favorecen la infección. METODOLOGÍA: Se realizó una búsqueda bibliográfica en Pub-Med con la siguiente estrategia de búsqueda: (("Bites, Human"[Mesh]) OR "Bites and Stings"[Mesh]) AND "Infection"[Mesh]. Se incluyeron 24 trabajos en la síntesis cualitativa escritos en lengua inglesa o española, casos clínicos o descriptivos y publicados entre los años 2000 y 2019. RESULTADOS: Las especies más frecuentemente aisladas fueron Capnocytophaga canimorsus en perros y Pasteurella multocida en gatos. La existencia de comorbilidades en el individuo, la mordedura en la mano, o la presencia de signos de alteración del estado general del individuo mordido fueron considerados como factores de riesgo para el desarrollo de la infección tras la mordedura. CONCLUSIONES: Todos los afectados por mordeduras animales deben recibir asistencia médica y considerar la administración de una pauta de profilaxis antimicrobiana con el fin de reducir el riesgo de shock séptico. Por otro lado, es importante advertir al laboratorio de microbiología de la naturaleza de las muestras clínicas obtenidas para alcanzar el mejor diagnóstico etiológico.

BACKGROUND: Bites caused by pets constitute 5% of the traumatic injuries registered in the emergency services. AIM: To know the main infectious agents present in dog and cat bites, both individually and jointly, in humans, as well as the predisposing factors that favor infection and its spread. METHODS: A bibliographic search was carried out in PubMed with the following search strategy: (("Bites, Human" [Mesh]) OR "Bites and Stings" [Mesh]) AND "Infection" [Mesh]. Twenty-four papers were included in the qualitative synthesis written in English or Spanish, clinical or descriptive cases and published between 2000 and 2019. Results: Most frequently isolated species were Capnocytophaga canimorsus in dogs and Pasteurella multocida in cats. The existence of comorbidities in the individual, the bite on the hand, or the presence of signs of alteration of the general state of the bitten individual were considered as risk factors for the development of infection after the bite. CONCLUSIONS: All patients with animal bites should receive medical assistance, and the administration of an antibiotic prophylaxis regimen should be considered to reduce the risk of septic shock. Besides, it is important to advise the microbiology laboratory of the nature of the clinical samples obtained in order to reach the best etiological diagnosis.

Agents of pneumonia in slaughtered pigs in southern Brazil

ABSTRACT: A high prevalence of pneumonic lesions has been reported to affect slaughtered pigs in southern Brazil. In order to identify which microorganisms have been causing those lesions, 30 pig lungs presenting pneumonic gross lesions were collected from five different slaughterhouses, totaling 150 lungs. Samples for bacterial isolation, molecular, histopathologic and immunohistochemistry (IHC) evaluation were taken from each lung. The pneumonic lesion scoring ranged from 1.53 to 2.83. The most frequent histopathological lesions found was the concomitant Influenza A virus (IAV) and Mycoplasma hyopneumoniae infection, corresponding to 55.3% (83/150), and Pasteurella multocida type A was isolated in 54.2% (45/83) of these cases. In 102 samples (68%), there was histopathologic suggestion of involvement of more than one infectious agent. M. hyopneumoniae was the most frequent agent associated with pneumonic lesions, being present in 92.1% (94/102) of the lungs with coinfections, followed by IAV in 89.2% (91/102). Besides the coinfections, IAV lesions were observed also in six samples without another pathogenic microorganism detected. A total of 46 samples with acute and subacute IAV suspected lesions in histopathological examination were assessed for IHC and real time RT-PCR for IAV. A total of 35% (16/46) of them were positive by IHC and 13% (6/46) by real time RT-PCR. Regarding M. hyopneumoniae, 79.3% (119/150) of samples were positive by qPCR and 84.9% (101/119) of them also presented M. hyopneumoniae suspected lesions in the histopathological examination. The results of this study suggest the importance of IAV in respiratory diseases in finishing pigs, even though this virus is more frequently reported in the nursery phase. In addition, our results emphasize the importance of lung coinfections in finishing pigs.

RESUMO: Lesões sugestivas de pneumonia são frequentemente encontradas em altas prevalências em suínos abatidos no sul do Brasil. Para identificar quais microrganismos causam essas lesões, foram coletados 30 pulmões de suínos com lesão macroscópica sugestiva de pneumonia em cinco frigoríficos diferentes, totalizando 150 pulmões. Amostras para isolamento bacteriano, avaliação molecular, histopatológica e imuno-histoquímica (IHC) foram coletadas de cada pulmão. O escore de lesão pulmonar variou entre 1,53 a 2,83. O achado histopatológico mais observado foi a lesão sugestiva de infecção concomitante pelo vírus Influenza A (IAV) e Mycoplasma (M.) hyopneumoniae, correspondendo a 55,3% (83/150), e em 54,2% (45/83) desses casos Pasteurella (P.) multocida tipo A foi isolado. Em 102 amostras (68%), houve lesão histopatológica sugestiva do envolvimento de mais de um agente infeccioso. M. hyopneumoniae foi o microrganismo mais frequente associado a lesões de pneumonia, estando presente em 92,1% (94/102) dos pulmões com coinfecções, seguido de IAV, que foi encontrado em 89,2% (91/102). Além das coinfecções, lesões de IAV foram observadas em mais seis amostras que não aparentavam envolvimento de outro agente infeccioso. Um total de 46 amostras com suspeita de lesão aguda e subaguda de IAV no exame histopatológico foram avaliadas para IHC e RT-PCR em tempo real para IAV e 35% (16/46) delas foram positivas por IHC e 13% (6/46) foram positivas por RT-PCR em tempo real. Com relação a M. hyopneumoniae, 79,3% (119/150) das amostras foram positivas por qPCR e 84,9% (101/119) delas também apresentaram lesões suspeitas de M. hyopneumoniae no exame histopatológico. Os resultados deste trabalho sugerem a importância do IAV como agente causador de pneumonias em suínos de terminação, embora esse vírus seja mais frequentemente relatado na fase de creche. Além disso, os achados deste trabalho demonstram a presença frequente de coinfecções pulmonares em suínos de terminação.

Agents of pneumonia in slaughtered pigs in southern Brazil / Agentes patogênicos de pneumonia em suínos abatidos no sul do Brasil

A high prevalence of pneumonic lesions has been reported to affect slaughtered pigs in southern Brazil. In order to identify which microorganisms have been causing those lesions, 30 pig lungs presenting pneumonic gross lesions were collected from five different slaughterhouses, totaling 150 lungs. Samples for bacterial isolation, molecular, histopathologic and immunohistochemistry (IHC) evaluation were taken from each lung. The pneumonic lesion scoring ranged from 1.53 to 2.83. The most frequent histopathological lesions found was the concomitant Influenza A virus (IAV) and Mycoplasma hyopneumoniae infection, corresponding to 55.3% (83/150), and Pasteurella multocida type A was isolated in 54.2% (45/83) of these cases. In 102 samples (68%), there was histopathologic suggestion of involvement of more than one infectious agent. M. hyopneumoniae was the most frequent agent associated with pneumonic lesions, being present in 92.1% (94/102) of the lungs with coinfections, followed by IAV in 89.2% (91/102). Besides the coinfections, IAV lesions were observed also in six samples without another pathogenic microorganism detected. A total of 46 samples with acute and subacute IAV suspected lesions in histopathological examination were assessed for IHC and real time RT-PCR for IAV. A total of 35% (16/46) of them were positive by IHC and 13% (6/46) by real time RT-PCR. Regarding M. hyopneumoniae, 79.3% (119/150) of samples were positive by qPCR and 84.9% (101/119) of them also presented M. hyopneumoniae suspected lesions in the histopathological examination. The results of this study suggest the importance of IAV in respiratory diseases in finishing pigs, even though this virus is more frequently reported in the nursery phase. In addition, our results emphasize the importance of lung coinfections in finishing pigs.(AU)

Lesões sugestivas de pneumonia são frequentemente encontradas em altas prevalências em suínos abatidos no sul do Brasil. Para identificar quais microrganismos causam essas lesões, foram coletados 30 pulmões de suínos com lesão macroscópica sugestiva de pneumonia em cinco frigoríficos diferentes, totalizando 150 pulmões. Amostras para isolamento bacteriano, avaliação molecular, histopatológica e imuno-histoquímica (IHC) foram coletadas de cada pulmão. O escore de lesão pulmonar variou entre 1,53 a 2,83. O achado histopatológico mais observado foi a lesão sugestiva de infecção concomitante pelo vírus Influenza A (IAV) e Mycoplasma (M.) hyopneumoniae, correspondendo a 55,3% (83/150), e em 54,2% (45/83) desses casos Pasteurella (P.) multocida tipo A foi isolado. Em 102 amostras (68%), houve lesão histopatológica sugestiva do envolvimento de mais de um agente infeccioso. M. hyopneumoniae foi o microrganismo mais frequente associado a lesões de pneumonia, estando presente em 92,1% (94/102) dos pulmões com coinfecções, seguido de IAV, que foi encontrado em 89,2% (91/102). Além das coinfecções, lesões de IAV foram observadas em mais seis amostras que não aparentavam envolvimento de outro agente infeccioso. Um total de 46 amostras com suspeita de lesão aguda e subaguda de IAV no exame histopatológico foram avaliadas para IHC e RT-PCR em tempo real para IAV e 35% (16/46) delas foram positivas por IHC e 13% (6/46) foram positivas por RT-PCR em tempo real. Com relação a M. hyopneumoniae, 79,3% (119/150) das amostras foram positivas por qPCR e 84,9% (101/119) delas também apresentaram lesões suspeitas de M. hyopneumoniae no exame histopatológico. Os resultados deste trabalho sugerem a importância do IAV como agente causador de pneumonias em suínos de terminação, embora esse vírus seja mais frequentemente relatado na fase de creche. Além disso, os achados deste trabalho demonstram a presença frequente de coinfecções pulmonares em suínos de terminação.(AU)

Antimicrobial resistance in bacteria isolated from pigs with respiratory clinical signs in Brazil / Resistência a antimicrobianos em bactérias isoladas de suínos com sinais clínicos respiratórios no Brasil

Antimicrobial resistance is a current and important issue to public health, and it is usually associated with the indiscriminate use of antimicrobials in animal production. This study aimed to evaluate the antimicrobial susceptibility profile in bacterial isolates from pigs with clinical respiratory signs in Brazil. One hundred sixty bacterial strains isolated from pigs from 51 pig farms in Brazil were studied. In vitro disk-diffusion method was employed using 14 antimicrobial agents: amoxicillin, penicillin, ceftiofur, ciprofloxacin, enrofloxacin, chlortetracycline, doxycycline, oxytetracycline, tetracycline, erythromycin, tilmicosin, florfenicol, lincomycin, and sulfadiazine/trimethoprim. The majority of isolates were resistant to at least one antimicrobial agent (98.75%; 158/160), while 31.25% (50/160) of the strains were multidrug resistant. Streptococcus suis and Bordetella bronchiseptica were the pathogens that showed higher resistance levels. Haemophilus parasuis showed high resistance levels to sulfadiazine/trimethoprim (9/18=50%). We observed that isolates from the midwestern and southern regions exhibited four times greater chance of being multidrug resistant than the isolates from the southeastern region studied. Overall, the results of the present study showed a great level of resistance to lincomycin, erythromycin, sulfadiazine/trimethoprim, and tetracycline among bacterial respiratory pathogens isolated from pigs in Brazil. The high levels of antimicrobial resistance in swine respiratory bacterial pathogens highlight the need for the proper use of antimicrobials in Brazilian pig farms.(AU)

A resistência antimicrobiana é uma questão atual e muito importante para a saúde pública, geralmente associada ao uso indiscriminado de antimicrobianos na produção animal. Diante disso, foi investigado o perfil de sensibilidade-antimicrobiana em isolados bacterianos de suínos com sinais clínicos respiratórios no Brasil. Foram estudadas 96 isolados provenientes de 51 granjas de suínos do Brasil. O método de disco-difusão foi empregado usando 14 antimicrobianos: amoxicilina, penicilina, ceftiofur, ciprofloxacina, enrofloxacina, clortetraciclina, doxiciclina, oxitetraciclina, tetraciclina, eritromicina, tilmicosina, florfenicol, lincomicina e sulfadiazina/trimetoprim. Streptococcus suis e Bordetella bronchiseptica foram os patógenos que apresentaram maiores níveis de resistência. Haemophilus parasuis apresentou altos níveis de resistência à sulfadiazina/trimetoprim (9/18=50%). Observou-se que isolados das regiões Centro-Oeste e Sul apresentaram quatro vezes mais chance de serem multirresistentes do que os isolados da região Sudeste. A maioria foi resistente a pelo menos um agente antimicrobiano (98,75%; 158/160) e 31,25% (50/160) das estirpes isoladas eram multirresistentes. No geral, os resultados do presente estudo mostraram grande nível de resistência à lincomicina, eritromicina, sulfadiazina/trimetoprim e tetraciclina entre patógenos respiratórios bacterianos isolados de suínos no Brasil. Os altos níveis de resistência antimicrobiana em patógenos bacterianos respiratórios em suínos reforçam a necessidade do uso criterioso de antimicrobianos na suinocultura brasileira.(AU)

Bacteriemia por Pasteurella multocida asociada al contacto con un animal doméstico / Pasteurella multocida bacteremia associated with contact with a domestic animal: case report

Resumen Pasteurella multocida es reconocida por ser una de las especies más frecuentemente aisladas en la microbiota oral de animales domésticos y salvajes; asociadas a infecciones de piel y tejidos blandos secundarias a mordeduras y rasguños. Las infecciones sistémicas son poco frecuentes, asociadas a la diseminación desde infecciones localizadas y a factores de riesgo asociados a inmunosupresión. Presentamos un caso de bacteriemia por Pasteurella multocida en un paciente de 88 años, relacionada con alimentos compartidos con su mascota; un mecanismo de producción de bacteriemia nunca antes descrito en la literatura médica.

Pasteurella species are known to be one of the most frequently isolated in oral microbiota of domestic and wild animals, because of that, they are associated with skin and soft tissues infections secondary to bites and scratches. Systemic infections are uncommon, but are associated with dissemination from localized infections and some risks factors related to immunosuppression. We report a case of Pasteurella multocida bacteremia in an 88 years old patient, associated with food sharing with his dog; a bacteremia mechanism never described before in the medical literature.

Antimicrobial susceptibility profile of historical and recent Brazilian pig isolates of Pasteurella multocida / Perfil de suscetibilidade a antimicrobianos de isolados históricos e contemporâneos de Pasteurella multocida de suínos no Brasil

Pasteurella (P.) multocida is the causative agent of pneumonic pasteurellosis in swine, which is commonly associated with the final stages of enzootic pneumonia or porcine respiratory disease complex. Although this syndrome is one of the most common and important diseases of pigs, data on antimicrobial susceptibility of P. multocida isolates are uncommon in Brazil. Therefore, the present study was carried out to determine and to compare antimicrobial susceptibility profile of Brazilian P. multocida isolated from pigs with lesions of pneumonia or pleuritis during two-time periods. Historical isolates (period of 1981 to 1997; n=44) and recent isolates (period of 2011 to 2012; n=50) were used to determine the MIC of amoxicillin, enrofloxacin, florfenicol and tetracycline by microbroth dilution. Florfenicol had the lowest level of resistance for both historical and recent isolates (0% and 6%, respectively), while tetracycline had the highest (20.5% and 34%, respectively). Multi-drug resistance (MDR) to amoxicillin/florfenicol/tetracycline was observed in 6% of recent isolates. There was a significant increase (p˂0.05) in resistance for amoxicillin and enrofloxacin in recent isolates compared with historic isolates (3.8% and 18%, respectively), most likely due to the selective pressure of antimicrobial usage to treat and prevent P. multocida infections. The results of this study showed an increase of isolates resistant to important drugs used in treatment of P. multocida infections in pigs, demonstrating the need for the implementation of rational use of antimicrobials in Brazilian swine industry.(AU)

Pasteurella (P.) multocida é o agente da pasteurelose pneumônica em suínos, a qual é comumente associada com o estágio final da pneumonia enzoótica suína ou complexo das doenças respiratórias dos suínos. Apesar de ser uma das doenças mais comuns e importantes na suinocultura, dados sobre suscetibilidade antimicrobiana de isolados de P. multocida são raros no Brasil. Dessa forma, o presente estudo foi realizado para determinar e comparar o perfil de suscetibilidade de isolados de P. multocida de suínos com lesões de pneumonia ou pleurite no Brasil durante dois períodos. Isolados históricos (período de 1981 a 1997; n=44) e contemporâneos (período de 2011 a 2012; n=50) foram usados para determinar a concentração inibitória mínima (CIM) de amoxicilina, enrofloxacina, florfenicol e tetraciclina através do teste de microdiluição em caldo. Florfenicol apresentou o menor nível de resistência para ambos os isolados históricos e contemporâneos (0% e 6%, respectivamente), enquanto que tetraciclina apresentou o maior nível de resistência (20.5% e 34%, respectivamente). Resistência a múltiplos antimicrobianos (amoxicilina, florfenicol e tetraciclina) foi observada em 6% dos isolados recentes. Foi observado aumento significativo (p˂0.05) na resistência a amoxicilina e enrofloxacina em isolados recentes comparado com isolados históricos (3.8% e 18%, respectivamente), provavelmente devido à pressão de seleção de antimicrobianos usados no tratamento e prevenção de infecções causadas por P. multocida. Os resultados deste trabalho demostraram o aumento de isolados resistentes a importantes drogas utilizadas no tratamento de infecções causadas por P. multocida em suínos, evidenciando a necessidade da implementação do uso racional de antimicrobianos na suinocultura brasileira.(AU)

In vitro treatment of lipopolysaccharide increases invasion of Pasteurella multocida serotype B:2 into bovine aortic endothelial cells

Pasteurella multocida serotype B:2 causes hemorrhagic septicemia in cattle and buffalo. The invasion mechanism of the bacterium when invading the bloodstream is unclear. This study aimed to characterize the effects of immunomodulatory molecules, namely dexamethasone and lipopolysaccharide, on the invasion efficiency of P. multocida serotype B:2 toward bovine aortic endothelial cells (BAECs) and the involvement of actin microfilaments in the invasion mechanism. The results imply that treatment of BAECs with lipopolysaccharide at 100 ng/mL for 24 h significantly increases the intracellular bacteria number per cell (p < 0.01) compared with those in untreated and dexamethasone-treated cells. The lipopolysaccharide-treated cells showed a significant decrease in F-actin expression and an increase in G-actin expression (p < 0.001), indicating actin depolymerization of BAECs. However, no significant differences were detected in the invasion efficiency and actin filament reorganization between the dexamethasone-treated and untreated cells. Transmission electron microscopy showed that P. multocida B:2 resided in a vacuolar compartment of dexamethasone-treated and untreated cells, whereas the bacteria resided in cellular membrane of lipopolysaccharide-treated cells. The results suggest that lipopolysaccharide destabilizes the actin filaments of BAECs, which could facilitate the invasion of P. multocida B:2 into BAECs.

Meningoencephalitis and pneumonia caused by Pasteurella multocida in rabbits / 대한수의학회지

Eight rabbits exhibited head tilt and subsequently died. At necropsy, three rabbits had crusty deposits in ears and four had reddish lungs. The main histopathological features were severe diffuse suppurative meningoencephalitis (75.0% of rabbits), fibrinopurulent pneumonia (37.5%), and otitis externa (37.5%). Pasteurella multocida (P. multocida) was isolated from brains, ears, and lungs. The capsular serogroups of the isolates were untypable. Based on histopathological features and bacterial analysis results, the rabbits were diagnosed as P. multocida infection. P. multocida infections might result in considerable economic loss in commercial rabbit production facilities in Korea.

Adjuvant activity of Pasteurella multocida A strain, Pasteurella multocida B strain and Salmonella typhimurium bacterial DNA on cellular and humoral immunity responses against Pasteurella multocida specific strain infections in Balb/c mice

Objective: To evaluate the effects of Pasteurella multocida (P. multocida) vaccines on the expression and release of antibodies, interleukin (IL)-6 and IL-12 by serum. Methods: Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA (AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA. Results: The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge (P<0.05). The highest release of these cytokines was obtained by P. multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum (P<0.05). Conclusions: Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines. These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.

Adjuvant activity of Pasteurella multocida A strain, Pasteurella multocida B strain and Salmonella typhimurium bacterial DNA on cellular and humoral immunity responses against Pasteurella multocida specific strain infections in Balb/c mice

Objective:To evaluate the effects of Pasteurella multocida (P. multocida) vaccines on the expression and release of antibodies, interleukin (IL)-6 and IL-12 by serum.Methods:Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA (AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA.Results:The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge (P<0.05). The highest release of these cytokines was obtained by P. multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum (P<0.05).Conclusions:Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines. These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.

Atypical Contact Lens Related Corneal Ulcer Caused by Pasteurella Multocida

A 55-year-old healthy lady withhistory of regular contact lens (CL) use presented with 10 days history of progressive left eye blurring of vision, redness and pain. There was good CL hygiene practiced with no history of swimming, trauma or contact with domestic pets. Left eye vision was hand movement and right eye was 1/60, pinhole 6/18. On the left eye, there was a central, oval-shaped corneal infiltrate with an overlying large epithelial defect and stromal oedema, with significant anterior chamber cells and fibrin. B-mode ultrasound showed no vitritis. Intensive topical benzylpenicillin 10000iu/ml and topical gentamycin 1.4% hourly, homatropine 2% three times daily, oral doxycycline and oral ascorbic acid were started. The gram stain results showed gram positive cocci growth. Her ulcerimproved with the treatment and preservative-free dexamethasone 0.1% once daily was commenced to reduce inflammation and scarring. Interestingly, culture was reported as Pasteurella maltocida, a gram negative bacilli sensitive to penicillin, and so treatment was continued until the ulcer completely healed. She had central corneal scarring with best corrected vision of 6/24 in the left eye but was not keen on further surgery to improve her vision. Although it has not been previously reported, Pasteurella multocidacan cause CL related corneal ulcer with severe anterior chamber inflammation. This diagnosis should be considered even if there is trivial contact or no history of exposure to domestic animals.

Anatomopathological pneumonic aspects associated with highly pathogenic Pasteurella multocida in finishing pigs / Aspectos pneumônicos anatomo-patológicos associados a Pasteurella multocida de alta patogenicidade em suínos de terminação

The bacterium Pasteurella multocida is a frequent cause of porcine respiratory disease complex in finishing pigs. Historically, the bacterium is recognized as an opportunistic agent, causing secondary bacterial pneumonia in pigs. Several Brazilian reports have suggested the ability of P. multocida to cause primary pulmonary infection that leads to the death of finishing pigs prior to slaughter. The aim of this study was to evaluate anatomopathological pulmonary findings associated with P. multocida infection that were obtained from animals with clinical respiratory disease and from animals at slaughter. Twenty-five lung samples from 14 herds of finishing pigs with acute clinical respiratory disease and 19 lungs collected at slaughter from a different set of 14 herds were studied. In all lung samples, bacterial isolation was performed, and only samples with pure P. multocida growth were included in the study. Gross and histopathological lesions were evaluated, as well as Influenza A, porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae co-infections. Pleuritis and pericarditis were more often observed in clinical samples (P<0.05). Moreover, there was a numerical trend indicating that pericarditis, lymphadenomegaly and cavity exudates were more often present in clinical samples. Thirteen lung samples were negative to M. hyopneumoniae, Influenza A and PCV2 by immunohistochemistry (IHC), with only P. multocida identified. In these cases, gross lesions such as pleuritis, pericarditis and lymphadenomegaly were always present, and no histologic lesions indicative of other agents such as Actinobacillus pleuropneumoniae, Actinobacillus suis or Haemophilus parasuis were observed. These findings suggest the ability of some P. multocida isolates to cause primary respiratory and systemic infection. However, in this study, it was not possible to determine specific virulence markers to explain these findings.

A bactéria Pasteurella multocida é causa frequente do Complexo de Doenças Respiratórias dos suínos em animais de terminação. Historicamente, a bactéria é reconhecida como agente oportunista, causando pneumonia bacteriana secundária. Diversos relatos brasileiros sugerem a habilidade da P. multocida de causar infecção pulmonar primária que leva a mortalidade de animais de terminação antes do abate. O objetivo deste estudo foi avaliar achados anatomopatológicos pulmonares associados com infecção por P. multocida, obtidas de animais acometidos clinicamente por doença respiratória e de animais ao abate. Avaliou-se 25 amostras de pulmão de 14 rebanhos obtidas de animais de terminação com sinais clínicos de doença respiratória aguda, e 19 pulmões coletados ao abate de 14 rebanhos diferentes. Em todos os pulmões, realizou-se isolamento bacteriano, e apenas amostras com crescimento puro de P. multocida foram incluídas no trabalho. Avaliou-se as lesões macro e microscopicamente, assim como co-infecções por Influenza A, Circovirus suíno tipo 2 (PCV2) e Mycoplasma hyopneumoniae. Pleurite e pericardite foram mais frequentemente observadas em amostras clinicas (P<0,05). Ainda, houve tendência numérica indicando a ocorrência de linfadenomegalia e exsudação cavitária, mais presentes em amostras clínicas. Treze amostras de pulmão foram negativas para M. hyopneumoniae, Influenza A e PCV2 por imunoistoquímica (IHQ), com identificação de apenas P. multocida. Nestes casos, lesões macroscópicas como pleurite, pericardite e linfadenomegalia foram sempre presentes, sem lesões histológicas indicativas de outros agentes como Actinobacillus pleuropneumoniae, Actinobacillus suis ou Haemophilus parasuis. Estes achados sugerem a habilidade de alguns isolados de P. multocida de causarem quadro respiratório primário e infecção sistêmica. No entanto, neste estudo, não foi possível determinar marcadores de virulência específicos para justificar tais achados.

Evaluation of the biofilm formation capacity of Pasteurella multocida strains isolated from cases of fowl cholera and swine lungs and its relationship with pathogenicity / Avaliação da capacidade de formação de biofilme por cepas de Pasteurella multocida isoladas de casos de cólera aviária e de pulmões de suínos e sua relação com a patogenicidade

Pasteurella multocida is a Gram-negative bacillus that causes economic losses due to the development of respiratory diseases in several animal species. Among the mechanisms of virulence, the formation of biofilms is an important factor for bacterial survival in hostile environments. Studies of biofilm formation by P. multocida are needed because P. multocida is an important pathogen involved in respiratory infections. However, in contrast to other microorganisms, few studies of biofilm formation have examined P. multocida. Studies comparing the pathogenicity of microbial strains as a function of their biofilm production capacity are also rare. Consequently, the aim of this study was to evaluate the biofilm formation capacity of 94 P. multocida strains isolated from cases of fowl cholera and from swine lungs on polystyrene plates. The associations of the biofilm formation capacity with the pathogenicity index (PI) in vivo and with the presence of four genes (screened by PCR) of the tad locus (tadB, tadD, tadE and tadG), described as adhesion markers, were also determined. Strains from both animal origins were able to form biofilms. However, most of the specimens (52.13%) were classified as weak producers, and more than 40% of the strains of P. multocida (40.42%) did not produce biofilms. There was no significant difference (p>0.05) in the degree of biofilm production between the two sources of isolation. Of the analyzed strains, 56.52% contained all four genes (tadB, tadD, tadE and tadG). The PI arithmetic mean of the strains classified as non-biofilm producers was significantly different (p<0.05) from the PI of moderate-producer strains. The PI of specimens classified as weak biofilm producers also differed significantly (p<0.05) from that of the moderate-producer strains. The results indicate that even though the P. multocida strains isolated from cases of fowl cholera and swine lungs formed biofilms on polystyrene surfaces, adhesion was usually weak. The genes tadB, tadD, tadE and tadG were not significantly associated (p>0.05) with the production of biofilms and with the origin of a given strain. Finally, low virulence strains may suggest a higher biofilm formation capacity on polystyrene plates.(AU)

Pasteurella multocida é um bacilo Gram negativo que ocasiona perdas econômicas, geralmente associadas a doenças respiratórias em diversas espécies animais. Entre os mecanismos de virulência existentes, a formação de biofilmes demonstra ser um importante fator para a proteção e para a sobrevivência bacteriana em ambientes hostis. Estudos relacionados à formação de biofilmes por P. multocida são necessários, uma vez que este é um importante patógeno envolvido em infecções respiratórias. Entretanto, ainda são poucos os estudos desenvolvidos nesta área, quando comparados com aqueles envolvendo outros microrganismos. Também são os raros os estudos que comparam a patogenicidade das cepas com a sua capacidade de produção de biofilme. Neste contexto, o objetivo deste estudo foi avaliar a capacidade de formação de biofilme em placas de poliestireno de 94 cepas de P. multocida isoladas de casos de cólera aviária e de pulmões de suínos, associando-se com o índice de patogenicidade (IP) in vivo e com a presença de quatro genes do locus tad (tadB, tadD, tadE e tadG), descritos como marcadores de adesão e pesquisados através de PCR. As cepas de ambas as origens foram capazes de formar biofilme. Contudo, a maioria dos exemplares (52,12%) foi classificada como fracamente produtora e mais de 40% das cepas de P. multocida (40,42%) não produziram biofilme. Não foi observada diferença estatística (p>0,05) quanto ao grau de produção de biofilme entre as duas origens de isolamento. 56,52% das cepas analisadas apresentaram os quatro genes (tadB, tadD, tadE e tadG) concomitantemente. O IP médio das cepas classificadas como não produtoras de biofilme apresentou diferença estatística (p˂0,05) em relação ao IP das cepas moderadamente produtoras. Os exemplares classificados como fracamente produtores de biofilme diferiram significativamente (p˂0,05) do grupo de cepas moderadamente produtoras. Os resultados obtidos indicaram que, apesar de as cepas de P. multocida isoladas de casos de cólera aviária e do pulmão de suínos apresentarem capacidade de formar biofilme em superfícies de poliestireno, a adesão ocorreu geralmente de forma fraca. Os genes tadB, tadD, tadE e tadG, pertencentes ao locus tad, não apresentaram associação significativa com a produção de biofilme e nem com a origem de isolamento da cepa. Por fim, observou-se que as cepas de menor patogenicidade apresentaram uma maior capacidade de formação de biofilme em placas de poliestireno.(AU)

Shock séptico de origen ginecológico por Pasteurella multocida / Septic shock of gynecological origin by Pasteurella multocida

La Pasteurella multocida es una gammaproteobacteria oportunista que produce una zoonosis caracterizada clínicamente por desarrollar cuadros mayoritariamente respiratorios como neumonía y rinitis atrófica, aunque las manifestaciones clínicas pueden derivar de la colonización de tejidos vascularizados y partes blandas de casi cualquier órgano, produciendo hemorragias, dermonecrosis, celulitis, meningitis, abscesos, septicemia, osteomielitis o endocarditis, entre otras. Este microorganismo se transmite a los humanos a través de mordeduras, arañazos o lesiones producidas por animales domésticos (especialmente gatos y perros), pero también salvajes. El diagnóstico es eminentemente clínico, apoyándose también en una anamnesis pormenorizada, y confirmándose mediante el crecimiento bacteriano en medios de cultivo como el agar sangre o el agar chocolate, de las muestras obtenidas de los pacientes afectados por esta rara infección. La antibioterapia con B-lactámicos durante 2 o 3 semanas, es la base terapéutica de este cuadro, aunque existe un alto porcentaje de pacientes con resistencia a los mismos, pudiendo necesitar terapias basadas en otros antibióticos como carbapenem, fluoroquinolonas o tetraciclinas. Las líneas de investigación más actuales están dando una gran importancia a los procedimientos de inmunización en animales domésticos, ya que por una parte son los principales vectores de transmisión y, por otro lado, la vacunación en humanos ha demostrado no ser efectiva, debido a la baja prevalencia de esta enfermedad en las personas expuestas. A continuación, presentamos el caso de una paciente con antecedentes de miomas uterinos y convivencia con gatos domésticos, que sufre un shock séptico por Pasteurella multocida, que tiene como principal foco infeccioso su útero miomatoso.

Pasteurella multocida is an opportunistic gammaproteobacteria which produces a zoonosis characterized clinically by developing majority respiratory pneumonia and atrophic rhinitis, even the clinical manifestations can be derived from the colonization of vascularized tissue and soft tissue of almost any organ, causing bleeding, dermonecrosis, cellulitis, meningitis, abscesses, sepsis, osteomyelitis or endocarditis, among others. This organism is transmitted to humans through bites, scratches or injuries caused by pets (especially cats and dogs), but also animals wild. The diagnosis is clinical, also leaning on a detailed anamnesis, and confirming through the bacterial growth in culture medium such as blood agar or chocolate agar samples from patients affected by this rare infection. With B-lactam antibiotic therapy for 2 or 3 weeks, is therapeutic base, although there is a high percentage of patients with resistance to them, and may need therapies based on other antibiotics as carbapenem, fluoroquinolones or tetracyclines. The lines of research are giving great importance to immunization procedures in domestic animals, since on the one hand are the main vectors of transmission and, on the other hand, vaccination in humans has proven to be not effective, due to the low prevalence of this disease in exposed persons. Then, present the case of a patient with a history of uterine fibroids and coexistence with domestic cats, suffering septic shock by Pasteurella multocida, which has as its main infectious focus your fibroid uterus.

Establishment and application of a multiplex PCR assay for four pathogens in laboratory animals / 中国比较医学杂志

The aim of this study is to establish a multiplex polymerase chain raction (PCR) to identify of four kinds of laboratory animal pathogens: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae and Klebsiella pneumoniae.Methods Specific primers were designed based on GenBank data.The multiplex PCR system was established through optimization of multiple PCR and detection of its specificity and sensitivity.This technique was used to test artificially infected samples and tracheal secretions of experimental animals (rat, mouse, guinea pig, rabbit, hamster), and comparing the detection results by this method and traditional detection test.Results Target bands of Pasteurella multocida (356 bp), Bordetella bronchiseptica (237 bp), Mycoplasma pneumoniae (266 bp), and Klebsiella pneumoniae (142 bp) were obtained, with a detection sensitivity of Klebsiella pneumoniae of 10 pg, and that of Pasteurella multocida, Bordetella bronchiseptica and Mycoplasma pneumoniae of 1 pg by this newly developed multiplex PCR assay.No target bands were observed from the non-specific pathogens of artificially infected samples.The tracheal secretions taken from 45 experimental animals (mice and rabbits) were tested with this new PCR assay, among which 15 cases of Klebsiella pneumonia and 9 cases of Pasteurella multocida were detected as positive, while all the results of traditional method and serological test were negative.Conclusions A simple, rapid, specific and highly sensitive multiplex PCR system has been successfully established.It is valuable for detection of Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae, and Klebsiella pneumoniae in laboratory animals.

Preliminary identification of the outer membrane protein H receptors of avian Pasteurella multocida / 中国人兽共患病学报

To identify the receptors for the outer membrane protein H (OmpH) of avian P.multocida,the membrane proteins of chicken embryo fibroblast (CEF) cells were separated by SDS-PAGE and analyzed by Ligand blot.The OmpH-binding protein was identified by MALDI-TOF mass spectrometry,and its distribution in the membrane proteins of different host esophageal mucosal cells was detected by Ligand blot,ELISA and immunofluorescence microscopy,respectively.Ligand blot analysis showed that a 49-kDa membrane protein of CEF cells bound to recombinant OmpH,and MALDI-TOF spectral results demonstrated that the OmpH-binding protein was ATP synthase β subunit.In addition,the OmpH receptor was present in the chicken and rabbit mucosal cell membranes,but was not detected in the bovine and swine mucosal cell membranes.The above results indicate that the OmpH receptor may be CEF cell-derived ATP synthase beta subunit.

Infecção por Pasteurella multocida em paciente cirrótico: relato de caso / Pasteurella multocida infection in a cirrhotic patient: a case report

Pasteurella multocida é uma bactéria que coloniza todo o trato gastrointestinal de cães e gatos. A infecção em humanos ocorre quando há arranhadura ou mordida por esses animais. Um subgrupo de pacientes com estado imunológico comprometido, como os cirróticos, possui certa propensão a desenvolver infecções sistêmicas por este germe. Neste relato descrevemos o caso de uma paciente com doença hepática alcoólica descompensada a partir de uma celulite em membro inferior direito (MID), peritonite bacteriana espontânea (PBE) e sepse, com culturas positivas para P. multocida. A despeito da antibioticoterapia e tratamento clínico adequado, o desfecho foi desfavorável. A paciente não apresentava história de mordida ou arranhadura por cães ou gatos.

Pasteurella multocida is a bacterium that colonises the entire gastrointestinal tract of cats and dogs. Infection in humans occurs when there is scratch or bite by these animals. A subgroup of patients with compromised immune status, such as cirrhosis, has a certain propensity to develop systemic infections by this germ. We describe a case of a patient with uncompensated alcoholic liver disease from a cellulitis in the lower limb, spontaneous bacterial peritonitis and sepsis, with positive cultures for P. multocida. Despite antibiotic therapy and appropriate clinical treatment outcome was unfavorable. The patient had no history of a bite or scratch by dogs or cats.