ISApl1, a novel insertion element of Actinobacillus pleuropneumoniae, prevents ApxIV-based serological detection of serotype 7 strain AP76

Actinobacillus pleuropneumoniae, a gram-negative rod of the Pasteurellaceae family, causes pleuropneumonia in pigs. Establishing A. pleuropneumoniae free herds is difficult due to the occurrence of persistently infected animals. The ApxIV toxin is expressed by A. pleuropneumoniae in vivo and an ELISA based on the toxin is used to detect infection and to differentiate between infected and vaccinated animals. In this study, we have identified a 1070bp insertion element of the IS30 family, designated ISApl1, in the A. pleuropneumoniae serotype 7 strain AP76. ISApl1 contains a 924bp ORF encoding a transposase, which is flanked by 27bp inverted repeats showing six mismatches. We investigated the occurrence of ISApl1 in other A. pleuropneumoniae strains, and its possible interference with virulence associated factors. Four insertion sites were identified in AP76: within the apxIVA toxin ORF, within a putative autotransporter adhesin ORF, upstream of a capsular polysaccharide biosynthesis gene cluster, and downstream of a beta-lactamase gene. ISApl1 is also present in some serotype 7 field isolates, but not in reference or field strains of other serotypes. In A. pleuropneumoniae AP76, the transposase gene is transcribed in vitro. The insertion in the apxIVA toxin gene remains stable after animal passage. Since this insertion should disrupt toxin expression, we tested 7 pigs infected with AP76 at day 21 post-infection. All were negative in the ApxIV ELISA but four out of seven were positive in an ApxII toxin ELISA. These results show that insertion elements can affect the detection of A. pleuropneumoniae infected animals.     

PCR specific for Actinobacillus pleuropneumoniae serotype 3

Serotypes 3 and 8 of Actinobacillus pleuropneumoniae, the aetiological agent of porcine pleuropneumonia, have been reported to predominate in the UK. Direct serotyping of isolates of the organism is typically determined by the immunological reactivity of rabbit serum to its surface polysaccharides, but the method has limitations, for example, cross-reactions between serotypes 3, 6 and 8. This study describes the development of a serotype 3-specific pcr, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The pcr test was evaluated on 266 strains of A pleuropneumoniae and 121 strains of other organisms, including all the major respiratory bacterial pathogens of pigs. The test was highly specific and sensitive and should be useful for differentiating strains of serotypes 3, 6 and 8, and in seroprevalence and epidemiological surveys in regions where serotype 3 is prevalent, such as the UK.     

Nitric Oxide Response in Critically III Dogs

Alterations in nitric oxide (NO)production may play a role in critical illness. Total serum nitrate/nitrite concentrations [SNN (uM/L)], the stable metabolites of NO, have been used as an indirect measure of NO in people, with increased concentrations reported in cases of critical illness. The relationship of nitric oxide (NO) to criticalillness in dgos is unknown. We tested the hypothesis that critically ill intensive care unit (ICU) canine illness in dogs is unknown. We tested the hypothesis that critically ill intesive care unit (ICU) canine patients would have increased SNN as compared to healthy dogs and non-critically ill dogs. An organ failure index score (OFI) was assigned to dogs admitted to the ICU to evaluate trends between disease severtiy and SNN. Critically ill dogs had significantly (p < 0.05) higher SNN (median 10.53) as compared to non-critically ill dogs (median 2.3) and healthy dogs (median 1.92). Critically ill dogs with the most severe disease (as based on OFI) had higher SNN concentrations. Survival of critically ill dogs with SNN of > 15 upon ICU admission (12% survival) was significantly less than survival of critically ill dogs with SNN ≤ 15 (91%) survival).l (Vet. Emerg. & Crit. Care, 9: 195–202, 1999)     

EVALUATION OF CANINE LUMBOSACRAL STENOSIS USING INTRAVENOUS CONTRAST-ENHANCED COMPUTED TOMOGRAPHY

The objective of this study was to evaluate intravenous contrast-enhanced computed tomography as a technique for predicting the within-level location(s) of compressive soft tissues in the canine lumbosacral spine. Pre-operative intravenous contrast-enhanced computed tomography of the L5-S3 vertebral levels was performed in 12 consecutive large breed dogs with lumbosacral stenosis. The images were evaluated for enhancement of soft tissues by two radiologists who were unaware of the surgical findings. For each within-level location (dorsal canal, ventral canal, right lateral recess, left lateral recess) enhancement was classified as present, absent or equivocal. The results were compared with the results of surgical exploration and histopathology of excised tissues. The positive predictive values of intravenous contrast-enhanced computed tomography for compressive soft tissues involving the dorsal canal, ventral canal and lateral recesses were 83%, 100%, and 81% respectively. Negative predictive values for compressive soft tissues involving these locations were 29%, 50%, and 40% respectively.     

RIGHT LEG MUSCLE ATROPHY AND OSTEOPENIA CAUSED BY RENAL ADENOCARCINOMA IN A COCKATIEL (MELOPSITTACUS, UNDULATUS)

A 7-year-old cockatiel presented with a 1 week history of right leg lameness. A renal adenocarcinoma invading and constricting the right ischiatic nerve resulted in disuse atrophy of the affected leg and radiographic evidence of osteopenia. This report illustrates the natural behavior, radiographic, and pathologic appearance of malignant renal tumors in birds.     

Habitat selection by spotted owls after a megafire reflects their adaptation to historical frequent-fire regimes

Context

Climate and land-use change have led to disturbance regimes in many ecosystems without a historical analog, leading to uncertainty about how species adapted to past conditions will respond to novel post-disturbance landscapes.

Objectives

We examined habitat selection by spotted owls in a post-fire landscape. We tested whether selection or avoidance of severely burned areas could be explained by patch size or configuration, and whether variation in selection among individuals could be explained by differences in habitat availability.

Methods

We applied mixed-effects models to GPS data from 20 spotted owls in the Sierra Nevada, California, USA, with individual owls occupying home ranges spanning a broad range of post-fire conditions after the 2014 King Fire.

Results

Individual spotted owls whose home ranges experienced less severe fire (<?5% of home range severely burned) tended to select severely burned forest, but owls avoided severely burned forest when more of their home range was affected (~ 5–40%). Owls also tended to select severe fire patches that were smaller in size and more complex in shape, and rarely traveled?>?100-m into severe fire patches. Spotted owls avoided areas that had experienced post-fire salvage logging but the interpretation of this effect was nuanced. Owls also avoided areas that were classified as open and/or young forest prior to the fire.

Conclusions

Our results support the hypothesis that spotted owls are adapted to historical fire regimes characterized by small severe fire patches in this region. Shifts in disturbance regimes that produce novel landscape patterns characterized by large, homogeneous patches of high-severity fire may negatively affect this species.

     

Sexing accuracy and indicators of maturation status in captive Murray cod Maccullochella peelii peelii using non-invasive ultrasonic imagery

Macroscopic- and histological-based assessments of gonad condition were compared with ultrasound images to determine the feasibility of this technology as a non-invasive diagnostic tool for identifying sex and assessing maturation status of Murray cod. Four age-classes (1+, 2+, 3+ and 6+ years), were sub-sampled at monthly intervals throughout their annual reproductive cycle and scanned with a 5 MHz linear transducer. An interpretation of sex was made from the resulting images and maximum cross-sectional gonad diameter and area were recorded. Fish were subsequently dissected to confirm gender, and the weights and maturation status of gonads determined and then compared with their respective image profile. Ovaries of females were usually a distinctive feature in ultrasound images, being particularly obvious in older and/or more developed fish. In contrast, the identification of male testis was more problematic. Nonetheless, identifying sex from ultrasound images was consistently achieved by recording the presence/absence of a female ovary (96% total sexing accuracy). Maximum cross-sectional ovary diameter and area were highly correlated with gonad weight (r² = 0.90 and 0.89, respectively) suggesting that indices of maturation status, comparable to the gonadosomatic index (GSI), can be obtained non-destructively from ultrasound scans of females. A less distinct relationship occurred between these dimensions and weight of testes (r² = 0.41). Significant increases (P < 0.05) in mean gonad index (GI, calculated from gonad diameter) occurred for most gonad development stages. However, differences in mean GI between maturation stages were confounded by phenotypic variability, indicating that GI may be limited to population level studies. Nevertheless, ultrasound images of ovaries at each development stage were visually distinctive and enabled qualitative evaluations of maturity, thereby complementing quantitative GI assessments. Repeated serial-monitoring of the same population using ultrasound appears to have great potential for tracking maturation-induced changes in broodfish.