Septicemia In Animals Research Articles

Development of ELISA-based diagnostic methods for the detection of haemorrhagic septicaemia in animals

Haemorrhagic septicaemia (HS) is an acute infection of cattle and buffaloes caused by the B:2 serotype of Pasteurella multocida. This disease is highly endemic in South Asia. In some peracute cases, there is 100% mortality in infected animals within a few hours of infection. Therefore, timely diagnosis of infection may contribute to its treatment and control to minimize economic losses. The current work reported the development of ELISA-based assays for the detection of anti-P. multocida antibodies and pathogen i.e. P. multocida. Owing to high immunogenicity, membrane proteins (MPs) extracted from local isolates of P. multocida serotype B:2 (PM1, PM2, and PM3) were employed as a potential diagnostic antigen for the development of indirect ELISA (i-ELISA) to detect HS antibodies in animals. MPs extracted from PM1, PM2 and PM3 isolates showed very low heterogeneity; hence MPs from the PM3 isolate were selected for the development of i-ELISA. The concentration of MPs (as coating antigen) of 3.13 μg/well and test sera dilution 1:100 was found to be optimal to perform i-ELISA. The developed method was validated through the detection of anti-P. multocida antibodies in sera of mice, immunized with MPs and formalin killed cells from the three local isolates (PM1, PM2 and PM3) of P. multocida. The significantly higher antibody titer in immunized mice was determined compared to unimmunized mice with the cut off value of 0.139. To detect P. multocida directly from the blood of infected animals, whole cell-based ELISA (cb-ELISA) assay was developed. A better detection signal was observed in the assay where bacterial cells were directly adsorbed on plate wells as compared to poly L-lysine (PLL) assisted attachment at a cell concentration of 106 CFU and 107 CFU respectively. The developed assays can be scaled up and potentially be used for the rapid detection of HS antibodies to gauge the immune status of the animal as well as vaccination efficacy and pathogen detection.

Use of black soldier fly (Hermetia illucens) prepupae amino acids as anti Aeromonas hydrophila enterotoxin in vivo

Aeromonas hydrophila is an opportunistic freshwater. These bacteria cause gastroenteritis and septicemia in animals and humans. Hemolysin and aerolysin, are important in the pathogenesis of A. hydrophila. Prepupae Black soldier fly (BSF) can be used as an antibacterial using its active substance against hemolysin and aerolysin. This study aimed to determine how the interaction of prepupae BSF amino acids with A. hydrophila enterotoxin in silico and protein level in various substrates in vivo. The study consisted of BSF larva of T1 (fed fruit waste), T2 (fed fermented fruit waste), T3 (fed tofu waste), T4 (fed fermented tofu waste), and T5 (fed fermented fruit waste and tofu waste). Data on the difference of protein level of prepupae among groups were analyzed statistically using the ANOVA test. The study showed that the highest protein content of BSF prepupae was found in treatment T3 dan T4. Protein docking analysis showed that L-arginine had the most hydrogen interaction (11 H-bonds) with aerolysin and 10 H-bonds against hemolysin, indicating an antibacterial role. The most favorable interacting residues of 17 amino acids against hemolysin were ARG73, ASP74, THR541, ALA523, and ASN483, while the residues of the active site against aerolysin were ASP92, ARG394, SER354, TYR348, ARG356, VAL396, PRO395, and ASP350. Amino could inhibit the hemolytic toxin of Aeromonas by interacting with binding site residues. The better the nutritional value of the substrate given to BSF larvae, the higher the protein content of BSF prepupae. Proteins from BSF prepupae can be antibacterial candidates against A. hydrophila.

Characterization of Streptococcus pluranimalium from a cattle with mastitis by whole genome sequencing and functional validation

BackgroundStreptococcus pluranimalium is a new member of the Streptococcus genus isolated from multiple different animal hosts. It has been identified as a pathogen associated with subclinical mastitis, valvular endocarditis and septicaemia in animals. Moreover, this bacterium has emerged as a new pathogen for human infective endocarditis and brain abscess. However, the patho-biological properties of S. pluranimalium remain virtually unknown. The aim of this study was to determine the complete genome sequence of S. pluranimalium strain TH11417 isolated from a cattle with mastitis, and to characterize its antimicrobial resistance, virulence, and carbon catabolism.ResultsThe genome of S. pluranimalium TH11417, determined by single-molecule real-time (SMRT) sequencing, consists of 2,065,522 base pair (bp) with a G + C content of 38.65%, 2,007 predicted coding sequence (CDS), 58 transfer RNA (tRNA) genes and five ribosome RNA (rRNA) operons. It contains a novel ISSpl1 element (a memeber of the IS3 family) and a Ф11417.1 prophage that carries the mef(A), msr(D) and lnu(C) genes. Consistently, our antimicrobial susceptibility test confirmed that S. pluranimalium TH11417 was resistant to erythromycin and lincomycin. However, this strain did not show virulence in murine pneumonia (intranasal inoculation, 107 colony forming unit – CFU) and sepsis (intraperitoneal inoculation, 107 CFU) models. Additionally, this strain is able to grow with glucose, lactose or galactose as the sole carbon source, and possesses a lactose-specific phosphoenolpyruvate-dependent phosphotransferase system (PTS).ConclusionsWe reported the first whole genome sequence of S. pluranimalium isolated from a cattle with mastitis. It harbors a prophage carrying the mef(A), msr(D) and lnu(C) genes, and is avirulent in the murine infection model.

Prevalence of plasmid-mediated quinolone resistance in Escherichia coli isolated from diseased animals in Taiwan.

Escherichia coli (E. coli) is a zoonotic pathogen that often causes diarrhea, respiratory diseases or septicemia in animals. Fluoroquinolones are antimicrobial agents used to treat pathogenic E. coli infections. In this study, 1,221 E. coli strains were isolated between March, 2011 and February, 2014. The results of the antimicrobial susceptibility testing showed a high prevalence of quinolone resistance. The antimicrobial resistance rates of these E. coli isolates to nalidixic acid (NAL) were 72.0% in swine, 81.9% in chickens, 81.0% in turkeys, 64.0% in ducks and 73.2% in geese. Among these isolates, the positive rate for the plasmid-mediated quinolone resistance (PMQR) determinant was 14.8% (181/1,221); the detection rate for qnrS1 was the highest (10.2%), followed by aac(6’)-Ib-cr (4.5%) and qnrB2 (0.3%). The quinolone-resistance determining regions (QRDRs) analysis for the PMQR-positive isolates showed that the strains with mutations at codon 83 or 87 in GyrA were resistant to NAL. To the best of our knowledge, this is the first report of occurrence of qnrB2, qnrS1 and aac(6’)-Ib-cr genes and high frequency (56.4%; 102/181) of mutation in gyrA or parC among PMQR-positive E. coli strains derived from diseased animals in Taiwan.

Rabbit Tularemia and Hepatic Coccidiosis in Wild Rabbit

Rabbit Tularemia and Hepatic Coccidiosis in Wild Rabbit

Phase variation of F165(1) (Prs-like) fimbriae from Escherichia coli causing septicaemia in animals.

Escherichia coli O115:F165 strains are associated with septicaemia in young pigs and synthesize fimbriae involved in virulence, designated as F165(1). F165(1) fimbriae belong to the P fimbrial family and are encoded by the foo gene cluster. The foo regulatory region of strain 5131 possesses characteristics similar to that of members of the P regulatory family, including papI and papB homologues, and two GATC sites separated by 102 bp, targets of differential Dam methylation. In wild-type strains, the synthesis of F165(1) is repressed by leucine and the fimbriae undergo phase variation. Immunofluorescence staining showed that phase variation of F165(1) results in a majority of cells (98%) in the ON phase, in contrast with phase variation of other members of this regulatory family, for which the majority of the cells are in the OFF state. Using a translational fusion in strain 5131 between phoA and fooA, encoding for the major structural subunit of F165(1), it was shown that leucine inhibits the OFF to ON switch and modulates the basal transcription of the foo operon.

気管支拡張症に併発した<I>Pasteurella multocida</I>感染症の1例

Pasteurella multocide (P. multocida), a small gram-negative bacillus, has been known to be the causative agent of hemorrhagic septicemia in animals. P. multocida infection in human was reported as skin abscess and/or septicemia after an animal bit or scratch. Pulmonary infections of P. multocida have been developed in the patients with chronic pulmonary diseases such as bronchiectasis. In Japan, however, P. multocida respiratory tract infections are rare. In this report, a 80-year-old female with bronchiectasis was admitted on August, 1985. She had a productive cough, hemosputum, and a low grade fever. The chest X-P on admission showed an atelectasis of the left middle lobe and severe bronchiectatic changes of the left lower lobe. P. multocida was isolated from her sputa. The chemotherapy of CTM resulted in clinical improvement. On May 1988, she complained of a productive cough and a low grade fever again. P. multocida was isolated from the sputum on several occasions in significant numbers (1 x 10(8)/ml). Recently, the cases of the chronic respiratory diseases have been increasing. We think, P. multocida is important and should be considered as a pathogen in the care of chronic pulmonary diseases.

Pasteurella multocida empyema.

PASTEURELLA multocida is a small, nonmotile, gram-negative, ovoid rod showing bipolar staining. Although it has been isolated from the respiratory and intestinal tracts of apparently normal cats, dogs, sheep, horses, fowl, swine, cattle and rats, it has long been known as the causative organism of hemorrhagic septicemia in animals.1 Human infections, though rarely reported, are probably far commoner than the literature leads one to believe.2 These infections may be of two types: a localized form resulting from an animal bite, usually that of a cat, with local abscess formation, sometimes resulting in osteomyelitis of the underlying bone; and a systemic . . .

Accessory Growth Factor Requirements of the Members of the Genus Pasteurella

Since preliminary work in this laboratory has shown that most of the members of the Pasteurella group could not be cultivated ini'synthetic mediums composed of amino acids, inorganic salts and glucose, a study of the accessory growth factor requirements of these organisms was undertaken. The organisms commonly included in this group are: (1) the hemorrhagic septicemia strains, which at present are designated according to their zoological source, such as Pasteurella avicida, Pasteurella boviseptica, etc., (2) Pasteurella pestis, (3) Pasteurella pseudotuberculosis and (4) Pasteurella tularense. Pasteurella hemolytica is the name suggested by Newsom and Cross1 for those organisms isolated from cases of hemorrhagic septicemia in animals, which nevertheless show certain physiological characteristics different from the typical forms (Past, avicida, Past, boviseptica, etc.). There has been some discussion as to whether or not the causative agent of tularemia belongs in the genus Pasteurella.