Intranasal instillation of Pasteurella multocida lipopolysaccharide in rabbits causes interstitial lung damage.

In order to characterize the in vivo lesions in the nasal cavities and lungs, twenty-eight rabbits were intranasally instilled with lipopolysaccharide (LPS) from P. multocida and then divided into seven groups according to euthanasia time. The nasal cavities and the lungs were processed for light microscopy, lectin histochemistry and transmission electron microscopy. Increased goblet cell activation and neutrophil infiltration were relevant changes in the nasal cavity. A predominantly interstitial pattern of diffuse alveolar damage and bronchopneumonic foci were the main lesions found in the lungs. LPS was found in the cytoplasm of ciliated cells, goblet cells, glandular cells, venular endothelial cells and neutrophils in the nasal cavity and in club cells, capillary endothelial cells and neutrophil in the lung. This study demonstrates that the LPS is able to cause lesions in the upper and lower respiratory tract, it binds to and is internalized by respiratory epithelial cells. Furthermore, it also traverses the intercellular spaces to reach the blood vessels, where it binds to and is internalized by neutrophil and red blood cells. These cells may then travel to the lungs where the LPS induces typical diffuse alveolar damage. This route of lung interstitial damage, to our knowledge, has not been described for this molecule or any known pathogen.

Pathogenomics insights for understanding Pasteurella multocida adaptation.

Pasteurella multocida is an important veterinary pathogen able to infect a wide range of animals in a broad spectrum of diseases. P. multocida is a complex microorganism in relation to its genomic flexibility, host adaptation and pathogenesis. Epidemiological analysis based on multilocus sequence typing, serotyping, genotyping, association with virulence genes and single nucleotide polymorphisms (SNPs), enables assessment of intraspecies diversity, phylogenetic and strain-specific relationships associated with host predilection or disease. A high number of sequenced genomes provides us a more accurate genomic and epidemiological interpretation to determine whether certain lineages can infect a host or produce disease. Comparative genomic analysis and pan-genomic approaches have revealed a flexible genome for hosting mobile genetic elements (MGEs) and therefore significant variation in gene content. Moreover, it was possible to find lineage-specific MGEs from the same niche, showing acquisition probably due to an evolutionary convergence event or to a genetic group with infective capacity. Furthermore, diversification selection analysis exhibits proteins exposed on the surface subject to selection pressures with an interstrain heterogeneity related to their ability to adapt. This article is the first review describing the genomic relationship to elucidate the diversity and evolution of P. multocida.

Pasteurella canis infection in a non-human primate black-tailed marmoset (Mico melanurus)-A case report.

This study reports the pathological and microbiological findings of pneumonia in a Mico melanurus caused by Pasteurella canis, confirmed for molecular analyses. It demonstrated the importance that wild species represent in the epidemiology of pasteurellosis in anthropic environments, when inserted into urban areas.

Surto de Pasteurelose por Pasteurella multocida tipo A em coelhos (Oryctolagus cuniculus) / Pasteurellosis outbreak due to Pasteurella multocida type A in rabbits (Oryctolagus cuniculus)

Background: Pasteurellosis is a common disease of cattle, pigs, and poultry, which rarely affects humans. In rabbits, therespiratory presentation of the disease is frequently reported. Clinical signs related to bronchopneumonia include sneezing, lung stertors, oculonasal discharge, dyspnea and cyanosis. Infection may lead to otitis, conjunctivitis, abscesses andsepsis. Furthermore, Pasteurella multocida infection may lead to sudden death without clinical manifestations. Reports ofpasteurellosis in rabbits are scarce in Brazil. Therefore, the objective of this article is to describe an outbreak of pasteurellosis with high mortality in a rabbity in the State of Santa Catarina, Brazil.Cases: Two adult rabbits were submitted for necropsy at the Veterinary Pathology Laboratory of the Instituto Federal Catarinense- Campus Concórdia, within an interval of twenty days. Herd was represented by 40 animals, of which six fattening rabbits andthree breeders died. Animals were kept in suspended cages with slatted floor. Clinical signs were represented by prostration,sneezing, and mucopurulent nasal discharge. In addition, wounds were observed in the distal portion of the limbs. Death occurredup to two days after the onset of clinical signs. Necropsies were performed and tissue samples were collected for histopathologic,immunohistochemical and microbiologic (bacterial culture and antibiogram) exams. At the necropsy, severe diffuse fibrinousexudate covering the pericardium sac, visceral and parietal pleural surfaces was noted, as well as multiple diaphragm adhesions.In addition, the lungs presented diffuse red coloration and showed multiple abscesses ranging from 0.3 to 1cm in diameter. Thenasal sinus and the tracheal mucosa showed diffuse reddening (rabbits 1 and 2). Abscesses up to 2 cm in diameter were observedin the mammary glands (rabbit 1), heart and kidneys (rabbit 2). The urinary bladder...(AU)

Surto de Pasteurelose por Pasteurella multocida tipo A em coelhos (Oryctolagus cuniculus) / Pasteurellosis outbreak due to Pasteurella multocida type A in rabbits (Oryctolagus cuniculus)

Background: Pasteurellosis is a common disease of cattle, pigs, and poultry, which rarely affects humans. In rabbits, therespiratory presentation of the disease is frequently reported. Clinical signs related to bronchopneumonia include sneezing, lung stertors, oculonasal discharge, dyspnea and cyanosis. Infection may lead to otitis, conjunctivitis, abscesses andsepsis. Furthermore, Pasteurella multocida infection may lead to sudden death without clinical manifestations. Reports ofpasteurellosis in rabbits are scarce in Brazil. Therefore, the objective of this article is to describe an outbreak of pasteurellosis with high mortality in a rabbity in the State of Santa Catarina, Brazil.Cases: Two adult rabbits were submitted for necropsy at the Veterinary Pathology Laboratory of the Instituto Federal Catarinense- Campus Concórdia, within an interval of twenty days. Herd was represented by 40 animals, of which six fattening rabbits andthree breeders died. Animals were kept in suspended cages with slatted floor. Clinical signs were represented by prostration,sneezing, and mucopurulent nasal discharge. In addition, wounds were observed in the distal portion of the limbs. Death occurredup to two days after the onset of clinical signs. Necropsies were performed and tissue samples were collected for histopathologic,immunohistochemical and microbiologic (bacterial culture and antibiogram) exams. At the necropsy, severe diffuse fibrinousexudate covering the pericardium sac, visceral and parietal pleural surfaces was noted, as well as multiple diaphragm adhesions.In addition, the lungs presented diffuse red coloration and showed multiple abscesses ranging from 0.3 to 1cm in diameter. Thenasal sinus and the tracheal mucosa showed diffuse reddening (rabbits 1 and 2). Abscesses up to 2 cm in diameter were observedin the mammary glands (rabbit 1), heart and kidneys (rabbit 2). The urinary bladder...

Outbreak of pasteurellosis in captive Bolivian squirrel monkeys (Saimiri boliviensis).

In September 2012, five Bolivian squirrel monkeys housed in a zoological park died within sequential several days without obvious clinical signs. In a necrospy, one monkey presented swelling of the kidney with multifocal white nodules in the parenchyma, and other two had pulmonary congestion. Histopathologically, multifocal bacterial colonies of gram-negative coccobacillus were found in the sinusoid of the liver in all monkeys examined (Nos.1-4). Additionally, purulent pyelonephritis, pneumonia and disseminated small bacterial colonies in blood vessels were observed. Immunohistochemically, the bacterial colonies from two monkeys were positive for P. multocida capsular serotype D. Based on these findings, these monkeys were diagnosed as septicemia caused by acute P. multocida infection.

Identificação de transcritos diferencialmente expressos por Pasteurella multocida em condições de privação de ferro / Identification of differentially expressed transcripts by Pasteurella multocida iron-starved condition

Ferro (Fe) é um elemento essencial e a capacidade de adquiri-lo in vivo têm sido descrita em diversos agentes patogênicos através de fatores de virulência. Análises de transcritos durante a privação de Fe tem sido descritos através da técnica de "microarray", entretanto a técnica de RNA-seq recentemente tem demonstrado resultados superiores. Neste trabalho, o isolado de Pasteurella multocida (Pm 16759) altamente patogênico em suínos foi cultivado em duas condições com diferentes concentrações de Fe (controle e privação) com o objetivo de analisar transcritos diferencialmente expressos. O RNA total das duas condições foi extraído e sequenciado através da plataforma de nova geração Ion Torrent. Os dados foram analisados no Software Ion Reporter(tm) e processados no programa Rockhopper. Foram obtidas 1.341.615 leituras com tamanho médio de 81pb, com 96% de alinhamento com o genoma de Pasteurella multocida subsp. multocida 3480 e 98,8% de acurácia. No mapeamento das leituras das duas condições, observou-se 2,652 transcritos e destes, 177 (6,7%) foram diferencialmente expressos, sendo 93 na condição controle (Fe+) e 84 na condição de privação (Fe-). Na condição de privação de Fe, o perfil de transcritos foram associados a função de transporte celular (fbp ABC, permease de alta afinidade com Fe2+/Pb2+ e proteína periplasmática de alta afinidade com Fe2+ ), reguladores transcricionais e proteínas hipotéticas. O perfil na condição controle (Fe+) apresentou transcritos diferencialmente expressos associados ao RNAs anti-sense (asRNA) e genes do metabolismo energético (fructose-1,6-bisfosfatase). O estudo comprovou que a restrição de Fe aumenta a expressão de genes envolvidos no transporte celular, reguladores transcricionais, proteínas hipotéticas e desconhecidas e permitiu ainda a identificação de novos genes como a permease de alta afinidade com Fe2+/Pb2+ e proteina periplasmática de alta afinidade com Fe2+ , que configuram uma possível via alternativa de absorção de Fe.(AU)

Iron (Fe) is an essential element and the ability to acquire it in vivo has been described in several pathogens as virulence factors. Global analyses of transcripts during iron deprivation have been described by microarray studies, however recently RNA-seq analysis showed superior results. The high pathogenic swine strain of Pasteurella multocida (BRMSA 1113) was grown in two conditions with different concentrations of Fe (control and deprivation) in order to analyze the differentially expressed transcripts. The total RNA of the two conditions was extracted and sequenced by new generation Ion Torrent plataform. Data were analyzed in Ion Reporter(tm) Software and processed in Rockhopper software. Sequence analysis shows 1,341,615 readings with median length of 81pb, with 96% of alignment to the reference genome Pasteurella multocida strain 3489, and 98.8% accuracy. Reads mapping to genome of P. multocida in these two conditions detected 2,652 transcripts, which 177 (6.7%) were differentially expressed, with 93 in the control condition (Fe+) and 84 provided with iron deprivation condition (Fe-). In condition (Fe-), differential expressed transcript profile were associated to function of cellular transport (fbpABC, high-affinity Fe2+/Pb2+ permease and periplasmic protein probably involved in hight-affinity Fe2+ ), transcptional regulators and hypothetical proteins. The control condition (Fe+) shows differential expressed transcripts profile associated to RNA anti-sense (asRNA) energetic metabolism genes (fructose-1,6-bisphosphatase). The study showed that the Fe restriction increases the expression of genes involved in cellular transport, transcriptional regulators, hypothetical and unknown proteins, and also allowed the identification of High-affinity Fe2+/Pb2+ permease e Periplasmic protein probably involved in high-affinity Fe2+, that constitute a possible alternative route for Fe absorption.(AU)

Identificação de transcritos diferencialmente expressos por Pasteurella multocida em condições de privação de ferro / Identification of differentially expressed transcripts by Pasteurella multocida iron-starved condition

Iron (Fe) is an essential element and the ability to acquire it in vivo has been described in several pathogens as virulence factors. Global analyses of transcripts during iron deprivation have been described by microarray studies, however recently RNA-seq analysis showed superior results. The high pathogenic swine strain of Pasteurella multocida (BRMSA 1113) was grown in two conditions with different concentrations of Fe (control and deprivation) in order to analyze the differentially expressed transcripts. The total RNA of the two conditions was extracted and sequenced by new generation Ion Torrent plataform. Data were analyzed in Ion Reporter(tm) Software and processed in Rockhopper software. Sequence analysis shows 1,341,615 readings with median length of 81pb, with 96% of alignment to the reference genome Pasteurella multocida strain 3489, and 98.8% accuracy. Reads mapping to genome of P. multocida in these two conditions detected 2,652 transcripts, which 177 (6.7%) were differentially expressed, with 93 in the control condition (Fe+) and 84 provided with iron deprivation condition (Fe-). In condition (Fe-), differential expressed transcript profile were associated to function of cellular transport (fbpABC, high-affinity Fe2+/Pb2+ permease and periplasmic protein probably involved in hight-affinity Fe2+ ), transcptional regulators and hypothetical proteins. The control condition (Fe+) shows differential expressed transcripts profile associated to RNA anti-sense (asRNA) energetic metabolism genes (fructose-1,6-bisphosphatase). The study showed that the Fe restriction increases the expression of genes involved in cellular transport, transcriptional regulators, hypothetical and unknown proteins, and also allowed the identification of High-affinity Fe2+/Pb2+ permease e Periplasmic protein probably involved in high-affinity Fe2+, that constitute a possible alternative route for Fe absorption.(AU)

Ferro (Fe) é um elemento essencial e a capacidade de adquiri-lo in vivo têm sido descrita em diversos agentes patogênicos através de fatores de virulência. Análises de transcritos durante a privação de Fe tem sido descritos através da técnica de "microarray", entretanto a técnica de RNA-seq recentemente tem demonstrado resultados superiores. Neste trabalho, o isolado de Pasteurella multocida (Pm 16759) altamente patogênico em suínos foi cultivado em duas condições com diferentes concentrações de Fe (controle e privação) com o objetivo de analisar transcritos diferencialmente expressos. O RNA total das duas condições foi extraído e sequenciado através da plataforma de nova geração Ion Torrent. Os dados foram analisados no Software Ion Reporter(tm) e processados no programa Rockhopper. Foram obtidas 1.341.615 leituras com tamanho médio de 81pb, com 96% de alinhamento com o genoma de Pasteurella multocida subsp. multocida 3480 e 98,8% de acurácia. No mapeamento das leituras das duas condições, observou-se 2,652 transcritos e destes, 177 (6,7%) foram diferencialmente expressos, sendo 93 na condição controle (Fe+) e 84 na condição de privação (Fe-). Na condição de privação de Fe, o perfil de transcritos foram associados a função de transporte celular (fbp ABC, permease de alta afinidade com Fe2+/Pb2+ e proteína periplasmática de alta afinidade com Fe2+ ), reguladores transcricionais e proteínas hipotéticas. O perfil na condição controle (Fe+) apresentou transcritos diferencialmente expressos associados ao RNAs anti-sense (asRNA) e genes do metabolismo energético (fructose-1,6-bisfosfatase). O estudo comprovou que a restrição de Fe aumenta a expressão de genes envolvidos no transporte celular, reguladores transcricionais, proteínas hipotéticas e desconhecidas e permitiu ainda a identificação de novos genes como a permease de alta afinidade com Fe2+/Pb2+ e [...](AU)

Outbreaks of avian cholera in Hope Bay, Antarctica.

During austral summers 1999-2000 and 2000-01, two outbreaks of avian cholera occurred in the Hope Bay area (63 degrees 24'S, 56 degrees 59'W), located on the tip of the Antarctic Peninsula. Eighty-six dead birds were found: five kelp gulls (Larus dominicanus), 36 skuas (Stercorarius sp.), and 45 Adelie penguins (Pygoscelis adeliae). The carcasses were studied using clinical, pathological, and microbiological criteria. Water samples from ponds where birds were settled and samples from 90 healthy birds also were analyzed during the second outbreak. Pasteurella multocida isolates were identified by biochemical tests, capsular type, somatic serotype, and susceptibility to nine antibiotics. Molecular subtyping was performed by ApaI and SmaI pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus (ERIC-PCR). In February 2000, mortality in skuas was 16% and 2% in kelp gulls. In the 2000-01 breeding season, mortality in south polar skuas was 47%, 24% in brown skuas, 1.4% in kelp gulls, and 0.01% in Adelie penguins. All birds had lesions of avian cholera. In kelp gulls the presentation was chronic, whereas skuas and penguins suffered subacute and acute disease, respectively. Fifty-five isolates recovered from dead birds and one from water were identified as P. multocida gallicida, type A:1. The strains presented a unique molecular pattern by PFGE and ERIC-PCR. A possible hypothesis to explain the origin of the outbreaks was that nonbreeder kelp gulls carried P. multocida gallicida to Hope Bay, and avian cholera was transmitted through water to skuas and penguins. This study reports avian cholera in new bird species, their potential role in the transmission of the disease, and the different responses of these species to the disease.

Avian cholera in a southern giant petrel (Macronectes giganteus) from Antarctica.

A southern giant petrel (Macronectes giganteus) was found dead at Potter Peninsula, King George Island, South Shetland, Antarctica. The adult male was discovered approximately 48 hr after death. Macroscopic and microscopic lesions were compatible with avian cholera and the bacterium Pasteurella multocida subsp. gallicida, serotype A1 was isolated from lung, heart, liver, pericardial sac, and air sacs. In addition, Escherichia coli was isolated from pericardial sac and air sacs. This is the first known report of avian cholera in a southern giant petrel in Antarctica.

Lechiguana (focal proliferative fibrogranulomatous panniculitis) in cattle.

Lechiguana is a disease of cattle characterized by large, hard, subcutaneous swellings that grow rapidly and result in death after 3-11 months in untreated animals. Cattle treated with antibiotics recover. The disease has been reported from five states in south and southeastern Brazil. Histologically, the lesion consists of focal proliferation of fibrous tissue infiltrated by plasma cells, eosinophils, lymphocytes and sometimes neutrophils. The primary lesion is an eosinophilic lymphangitis, which results in eosinophilic abscesses, with occasional rosettes containing bacteria in their centres. Much experimental and epidemiological evidence, reviewed in this article, supports the suggestion that lechiguana is caused by an association of Pasteurella granulomatis (syn: Mannheimia granutomatis) and Dermatobia hominis.

Filamentous intranuclear inclusion bodies in psittacine birds. A structural and ultrastructural study.

This paper concerns a disease affecting a group of African grey parrots, which involves intranuclear inclusion bodies composed of filamentous material. The disease was characterized by either sudden death or death within 2-3 days from onset of non-specific symptoms. At necropsy, gross lesions included enlarged liver, mild hepatic congestion and focal necrosis. Samples from five birds were fixed in 10% formol and routinely processed for light and electron microscopy. In four birds, numerous hepatocytes displayed an enlarged nucleus, with peripheral margination of chromatin; the nucleus was partially or wholly filled by a basophilic inclusion body. In the remaining bird, inclusion bodies were acidophilic and completely filled the nucleus; nuclear enlargement was less evident than in the other birds. At ultrastructural examination, and in both types of IIB, nuclei contained looped filaments but no evidence of viral structures. However, virion-like structures were observed in the cytoplasm of some hepatocytes.

Adherence of Pasteurella multocida to rabbit respiratory epithelial cells in vitro.

Adult clinically healthy New Zealand rabbits were sampled bacteriologically to detect carriers and non-carriers of Pasteurella multocida. Both groups of rabbits were killed separately to obtain samples of nasal, buccal, pharyngeal and tracheal epithelial cells. The cells were tested for adherence in vitro to 18 isolates of P. multocida from healthy and sick rabbits, from ovine, bovine, cat and swine. The number of bacteria adhered per cell up to 25 cells per preparation were registered. Analysis of variance was used to interpret the significance of results. Adherence of P. multocida was significantly higher to carrier rabbit cells than to non-carrier rabbit cells. Bacterial isolates from rabbits were more adherent to rabbit cells than to isolates from other species. The frequency was higher to buccal and pharyngeal cells than to nasal and tracheal cells. Isolates from healthy animals adhered better to rabbit cells than isolates from sick animals, except the isolates from sick animals which adhered better to nasal cells of non carriers than did isolates from healthy rabbits.

Pasteurella haemolytica infection in a Goeldie's monkey.

An adult male Callimico goeldii died spontaneously. At necropsy, small whitish foci were found randomly distributed on the liver surface. Histologically, the foci were composed of mixed inflammatory cells with predominant polymorphonuclear cell infiltration and central areas of necrosis. Microbiological cultures revealed a Gram-negative coccoid-bacilli with bipolar staining. Biochemical analysis revealed that the microorganism was Pasteurella haemolytica.

Bovine focal proliferative fibrogranulomatous panniculitis (Lechiguana) associated with Pasteurella granulomatis.

In southern Brazil, cattle are affected by a disease known locally as Lechiguana and characterized by large subcutaneous swellings. Eighteen cases were examined clinically; 17 of the cattle had a single swelling, and one had two swellings. In 14 of the 18 cases, the swellings were located over the scapula and adjacent regions. The subcutaneous masses reached maximum dimensions of 45 x 50 cm, with heights above the skin surface of 5-25 cm. Growth was rapid, often taking place in 15 to 60 days. Histologically, all lesions were focal proliferative fibrogranulomatous panniculitis and consisted of focal proliferation of fibrous tissue that was infiltrated by plasma cells, eosinophils, lymphocytes, and sometimes neutrophils. An eosinophilic lymphangitis was also present, which sometimes resulted in the destruction of the lymphatics and the formation of eosinophilic microabscesses. Small granulomas, sometimes containing radiating clubs, and Splendore-Hoeppli material were present in the regional lymph node. Pasteurella granulomatis was isolated from the subcutaneous masses of 14 of the 18 natural cases. All 11 of these cases recovered following treatment with 3 g of chloramphenicol daily for 5 days. Untreated animals died. Because the area of anatomic distribution is similar to that infested by Dermatobia hominis, we postulate that this insect may transmit the causative agent. In one steer, a subcutaneous injection of P. granulomatis caused a large subcutaneous swelling consisting of interlacing bundles of collagen infiltrated by neutrophils, eosinophils, and some lymphocytes. Microabscesses, but not lymphangitis and granulomas, were detected. In all 11 cattle inoculated either intramuscularly or subcutaneously with P. granulomatis, purulent abscesses were produced at the sites of the injection, and P. granulomatis was recovered from all lesions.

Leishmania braziliensis: effects of bacteria (Staphylococcus aureus and Pasteurella multocida) on the developing cutaneous leishmaniasis lesion in the golden hamster.

Experimentally induced lesions of cutaneous leishmaniasis and the effect of concurrent bacterial infection on the development of these lesions were studied in the golden hamster. Male outbred golden hamsters received intradermal injections at the base of the tail with approximately 10(7) promastigotes of Leishmania braziliensis panamensis, or promastigotes combined with Staphylococcus aureus or Pasteurella multocida or both, bacteria only, or sterile Eagle's minimal essential medium (MEME). The size of the resulting lesions was measured at least twice each week. Hamsters were killed at postinoculation Days 6, 13, 20, 27, 41, or 48, and each lesion was measured, aseptically excised, and bisected; half was used for bacteriologic culture and the other half was prepared for light microscopic examination. Lesions resulting from L. b. panamensis alone progressed from initial erythema to a granulomatous nodule and finally to a necrotic granuloma, often capped by a crateriform ulcer. Lesions resulting from a suspension of L. b. panamensis with added S. aureus or S. aureus and P. multocida, were initially larger, more erythemic and contained a greater proportion of neutrophils up to postinoculation Days 14-21 than did lesions resulting from L. b. panamensis alone. Concurrent infections with bacteria such as S. aureus and P. multocida had little effect on the development of ulcerating characteristics of lesions, but when S. aureus was present it appeared to enhance the severity of the early lesions. Between postinoculation Days 14-28, lesions produced by L. b. panamensis, with or without added bacteria had similar developmental progression of sufficient size for optimal testing of antileishmanial compounds.