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Abstract: The objective of the present study was to evaluate the effect of immunomodulatory therapy (glucocorticoids (GC) and mycobacterium cell wall extract (MCWE)) on the endometrial gene expression of inflammatory cytokines in susceptible mares with induced infectious endometritis. Endometrial gene expression of pro- and anti-inflammatory cytokines; interleukin (IL)-1β, IL-6, IL-8, IL-10, tumor necrosis factor (TNF)-α, IL-1 receptor antagonist (ra), acute phase protein (APP) serum amyloid A (SAA) and clinical parameters were evaluated. Five mares were classified as susceptible to persistent endometritis based on their endometrial histopathology and ability to clear an induced uterine inflammation. To investigate the effect of immunomodulatory therapy, the mares were inoculated with 105 colony forming units (CFU) Escherichia coli in three consecutive estrus cycles in a modified cross-over study design. Thus, each mare served as its own control and the treatment type was performed in randomized order. The effect of treatment with MCWE (1.5 mg Settle IV), dexamethasone (0.1 mg per kg IV) or no treatment was investigated. All mares were free from uterine inflammation before each E. coli inoculation. Endometrial biopsies were recovered 3, 24 and 72 h post inoculation. Relative gene-expression analyses were performed by quantitative reverse transcriptase PCR (qRT-PCR). Endometrial gene expression of inflammatory cytokines was modulated by administration of GC. Expression of proinflammatory cytokines (IL-1β, IL-6, IL-8) and SAA was significantly lower in the GC treated group late in the study period (72 h) compared to “no treatment” and MCWE treatment. Increased expression of the anti-inflammatory cytokine IL-10 was observed 3 and 24 h after E. coli infusion and GC treatment. A significant decrease of SAA expression was observed after MCWE treatment compared to “no treatment”. MCWE and GC treatment had a significant effect on the clearance of uterine pathogens and number of mares retaining fluid after E. coli infusion. The results of the current investigation suggest that GC is capable of effectively modulating the innate immune response to induced infectious endometritis in susceptible mares. [Copyright &y& Elsevier]

Threatened by Devil Facial Tumor Disease, the Tasmanian devil populations are vulnerable and decreasing. Additionally, the devils' biting behaviour elevates their risk of acquiring bite wound infections caused by members of the bacterial Pasteurellaceae family that are natural inhabitants of the oral microbiota. In medical management of such bite wounds, antimicrobial susceptibility profiles are crucial. Prior to this investigation, no available data on minimal inhibitory concentration ( MIC) values existed. A total of 26 isolates obtained from the oral cavity of 26 healthy Tasmanian devils were tested for their antimicrobial susceptibility by broth micro dilution. Most prominently, high MIC values for clindamycin (≥4 μg ml−1), gentamicin (≥8 μg ml−1) and amikacin (≥32 μg ml−1), were observed for 92, 77 and 73% of the strains tested respectively. This study may be used as a guideline for antimicrobial therapy against bite wound infections caused by Pasteurellaceae originating from the oral cavity of Tasmanian devils. Significance and Impact of the Study Tasmanian devils' aggressive behaviour makes bite wounds in fellow devils and human caretakers a common entity. Pasteurellaceae bacteria are common inhabitants of the oral microbiota of Tasmanian devils and a likely cause of bite wound infections. Here, for the first time, we report antimicrobial sensitivity profiles from a broad collection of Pasteurellaceae isolates obtained from the oral cavity of Tasmanian devils. Low MIC values were observed for the majority of the 22 antimicrobial agents included, yet nearly all strains were tolerant to clindamycin and the aminoglycosides. The work can serve as a guide for clinicians involved in treatment of bite wounds inflicted by devils in animals and humans. [ABSTRACT FROM AUTHOR]

Endometritis in horses caused by Streptococcus equi subspecies zooepidemicus ( S. zooepidemicus ) may be underdiagnosed due to traditional diagnostic methods lacking sensitivity and specificity. We serendipitously identified a bacterial growth medium (bActivate) that appeared capable of inducing growth of dormant S. zooepidemicus , which subsequently allowed detection by standard diagnostics. To assess the effect of bActivate we compared its ability to activate dormant S. zooepidemicus in a group of potentially infected subfertile mares with phosphate-buffered saline (PBS). All mares had to test negative for S. zooepidemicus on a low-volume uterine lavage , be negative on endometrial cytology and without clinical signs of endometritis to be included in the investigation. The mares were instilled with bActivate or PBS in the uterus. Growth of S. zooepidemicus was induced by bActivate in 64% (16/25) and PBS in 8% (1/12) of the mares, respectively ( p < 0.002). In vitro studies supported that some strains of S. zooepidemicus were able to form persister cells tolerating 32-times of the minimal inhibitory concentration of penicillin compared to normal growing cells. Persister cells had not acquired penicillin resistance, but seemed to tolerate the antimicrobial due to dormancy. This is, to our knowledge, the first description of controlled growth induction of dormant bacteria from a subclinical infection. Moreover we demonstrated how endometritis can origin from a reservoir of dormant bacteria residing within the endometrium, and not only as an ascending infection. Further studies should aim at determining the prevalence of dormant S. zooepidemicus, impact of activation on diagnostic and treatment efficacy, uterine health and mare fertility. [ABSTRACT FROM AUTHOR]

Detailed longitudinal studies of the genetic stability of Pasteurella multocida ssp. multocida , the cause of fowl cholera, have not previously been carried out. Consequently, the aim of the present study was to provide detailed information on the genetic stability and diversity of P. multocida ssp. multocida in poultry flocks over time, enabling new insights into the molecular epidemiology of this important poultry pathogen. Longitudinal investigations of the rate and causes of mortality were carried out on two free-range layer farms (A and B) over a period of 11 months. The total mortality of two flocks, A 1 and A 2 , on farm A were 62 and 91%, respectively, while the total mortality of a single flock B 1 on farm B was 6%. Postmortem examinations were performed on 708 layers from flocks A 1 and A 2 and in 159 from flock B 1 . Fowl cholera was the main cause of mortality on both farms. Pasteurella multocida isolates recovered from layers on both farms were characterized phenotypically and genotypically, and 322 isolates were identified as P. multocida ssp. multocida . The genetic diversity of 99 isolates from farm A and 31 from farm B was characterized by restriction endonuclease analysis and amplified fragment length polymorphism analysis. The isolates on each farm had a unique restriction endonuclease analysis and amplified fragment length polymorphism analysis type, suggesting a single introduction of a successful clone. Furthermore the clone on farm A was identical to clones previously isolated from outbreaks in the avifauna of Denmark in 1996 and 2001 and in Sweden in 1998. This study provides convincing evidence for the clonal stability of outbreak clones of P. multocida . Stabilité clonale de Pasteurella multocida chez des poules pondeuses élevées en plein air affectées par le choléra aviaire Jusqu'à présent, des études longitudinales et détaillées de la stabilité génétique de Pasteurella multocida ssp. multocida, l'agent responsable du choléra, n'avaient été réalisées. En conséquence, le but de cette étude a été de fournir des informations détaillées sur la stabilité génétique et la diversité de P. multocida ssp. multocida dans les troupeaux de volailles dans le temps, permettant de fournir de nouveaux aperçus sur l'épidémiologie moléculaire de cet agent pathogène important pour les volailles. Les investigations longitudinales des taux et des causes de mortalité ont été réalisées dans deux élevages de poules pondeuses élevées en plein air (A et B) sur une période de 11 mois. Les mortalités totales de deux troupeaux A1 et A2 appartenant à l'élevage A ont été respectivement 62% et 91%, alors que la mortalité totale du seul troupeau B1 de l'élevage B a été de 6%. Les examens post mortem ont été réalisés sur 708 pondeuses des troupeaux A1 et A2 et sur 159 du troupeau B1. Le choléra aviaire a été la principale cause de mortalité au niveau des deux élevages. Les souches de Pasteurella multocida isolées à partir des pondeuses des deux fermes ont été caractérisées sur les plans phénotypique et génétique et 322 souches ont été identifiées comme P. multocida ssp. multocida . La diversité génétique de 99 souches de l'élevage A et de 31 souches de l'élevage B a été caractérisée par l'analyse de restriction enzymatique (REA) et l'analyse du polymorphisme de taille d'un fragment amplifié (AFLP). Les souches de chaque élevage ont présenté un seul type de REA et AFLP suggérant l'introduction d'un clone unique. De plus, le clone de l'élevage A était identique aux clones isolés précédemment de cas pathologiques de l'avifaune au Danemark en 1996 et en 2001 ainsi qu'en Suède en 1998. Cette étude fournit une évidence convaincante de la stabilité clonale des souches épidémiques de P. multocida . Klonale Stabilität von Pasteurella multocida bei von Gefügelcholera betroffenen Freilandhühnern Bislang sind detaillierte Langzeitstudien zur genetischen Stabilität von Pasteurella multocida ssp. multocida , dem Erreger der Geflügelcholera nicht durchgeführt worden. Somit war das Ziel der vorliegenden Studie, detaillierte Informationen über die genetische Stabilität und Vielfalt von P. multocida ssp. multocida in Geflügelherden über einen langen Zeitraum zu gewinnen und damit neue Einsichten in die molekulare Epidemiologie dieses bedeutsamen Krankheitserregers beim Geflügel zu ermöglichen. In zwei Legehennenauslaufhaltungen (A und B) wurden über einen Zeitraum von 11 Monaten Langzeituntersuchungen zu den Mortalitätsraten und ihren Ursachen durchgeführt. Die Mortalität in den beiden Herden A 1 und A 2 auf der Farm A betrug insgesamt 62 bzw. 91 %, während in der einen Herde B 1 auf der Farm B eine Mortalität von 6 % erreicht wurde. Bei 708 Legehennen aus den Herden A 1 und A 2 und bei 169 Hennen aus der Herde B 1 wurden Sektionen vorgenommen. Auf beiden Farmen war die Geflügelcholera die Haupttodesursache. Pasteurella multocida -Isolate aus Legehennen beider Farmen wurden phänotypisch und genotypisch charakterisiert und 322 Isolate wurden als Pasteurella multocida ssp. multocida identifiziert. Die genetische Diversität von 99 Isolaten aus Farm A und 31 aus Farm B wurde mittels Restriktionsendonukleaseanalyse (REA) und amplifizierter Framentlängenpoly-morphismusanalyse (AFLP) charakterisiert. Die Isolate auf jeder Farm gehörten jeweils zu nur zu einem einzigen REA- und AFLP-Typ, was auf eine einmalige Einschleppung eines erfolgreichen Klons schließen lässt. Außerdem war der Klon von Farm A identisch mit Klonen, die bereits bei Ausbrüchen in Geflügelbeständen 1996 und 2001 in Dänemark und 1998 in Schweden isoliert worden waren. Diese Studie erbringt einen überzeugenden Beweis der klonalen Stabilität von bei Krankheitsausbrüchen isolierten P. multocida -Klonen. Estabilidad de los clones de Pasteurella multocida en ponedoras en extensivo afectadas por cólera aviar Estudios longitudinales detallados de la estabilidad genética de Pasteurella multocida ssp. m ultocida, agente causal del cólera aviar, no se han desarrollado con anterioridad. Por lo tanto, el objetivo de este estudio fue obtener información detallada en el tiempo de la estabilidad genética y de la diversidad de P. multocida ssp. Multocida en lotes de aves, obteniendo nuevas ideas acerca de la epidemiología molecular de este importante patógeno aviar. Se llevaron a cabo estudios longitudinales del porcentaje y de las causas de mortalidad en dos granjas de ponedoras en extensivo (A y B) durante un período de 11 meses. La mortalidad total en ambos lotes de la granja A, A 1 y A 2 , fue del 62% y del 91% respectivamente, mientras que la mortalidad total del único lote en la granja B, el B 1 , fue del 6%. Se realizaron estudios postmortem en 708 gallinas ponedoras de los lotes A 1 y A 2 , y 159 del lote B 1 . El cólera aviar fue la principal causa de mortalidad en ambas granjas. Se caracterizaron fenotípica y genotípicamente aislamientos de Pasteurella multocida obtenidos de ponedoras de ambas granjas, y 322 aislamientos fueron identificados como P. multocida ssp. multocida . Se caracterizó la diversidad genética de 99 aislamientos de la granja A y 31 de la granja B mediante análisis de la restricción con endonucleasas (REA) y análisis del polimorfismo de la longitud de los fragmentos amplificados (AFLP). Los aislamientos de cada granja mostraron patrones característicos de REA y AFLP, sugiriendo una única entrada de un clon exitoso. Además el clon de la granja A era idéntico a clones aislados previamente de brotes en la avifauna de Dinamarca en 1996 y 2001y en Suecia en 1998. Este estudio proporciona evidencias de la estabilidad de los clones causantes de brotes de P. multocida. [ABSTRACT FROM AUTHOR]