Data_Sheet_1_Nephrocalcinosis in farmed salmonids: diagnostic challenges associated with low performance and sporadic mortality.PDF

Disease conditions that involve multiple predisposing or contributing factors, or manifest as low performance and/or low-level mortality, can pose a diagnostic challenge that requires an interdisciplinary approach. Reaching a diagnosis may also be limited by a lack of available clinical profile parameter reference ranges to discriminate healthy fish from those affected by specific disease conditions. Here, we describe our experience investigating poorly performing rainbow trout (Oncorhynchus mykiss) in an intensive recirculation aquaculture, where reaching a final diagnosis of nephrocalcinosis was not as straightforward as one would wish. To list the issues making the diagnosis difficult, it was necessary to consider the creeping onset of the problem. Further diagnostic steps needed to ensure success included obtaining comparative data for fish blood profiles and water quality from both test and control aquacultural systems, excluding infections with salmonid pathogenic agents and evaluating necropsy findings. Major events in the pathophysiology of nephrocalcinosis could be reconstructed as follows: aquatic environment hyperoxia and hypercapnia → blood hypercapnia → blood acid-base perturbation (respiratory acidosis) → metabolic compensation (blood bicarbonate elevation and kidney phosphate excretion) → a rise in blood pH → calcium phosphate precipitation and deposition in tissues. This case highlights the need to consider the interplay between water quality and fish health when diagnosing fish diseases and reaching causal diagnoses.</p

Additional file 2: of White-nose syndrome detected in bats over an extensive area of Russia

Table S1. Bat species from Palearctic and Nearctic regions with confirmed WNS or Pseudogymnoascus destructans infection. Summarized from [10, 16, 19, 21, 40, 47]. (XLSX 13 kb

Additional file 1: of White-nose syndrome detected in bats over an extensive area of Russia

Figure S1. Infection intensity, measured as fungal load in nanograms on a log10 scale, for particular regions. Explanation: mid-point = median; box = inter-quartile range; whiskers = non-outlier minimum/maximum range; dots = outliers; stars = extremes. (DOCX 21 kb

Additional file 3: of White-nose syndrome detected in bats over an extensive area of Russia

Table S2. Datasets used and analyzed during the current study and not included in Tables 1, 2 and 3. (XLSX 31 kb

White-Nose Syndrome Fungus: A Generalist Pathogen of Hibernating Bats

<div><p>Host traits and phylogeny can determine infection risk by driving pathogen transmission and its ability to infect new hosts. Predicting such risks is critical when designing disease mitigation strategies, and especially as regards wildlife, where intensive management is often advocated or prevented by economic and/or practical reasons. We investigated <i>Pseudogymnoascus [Geomyces] destructans</i> infection, the cause of white-nose syndrome (WNS), in relation to chiropteran ecology, behaviour and phylogenetics. While this fungus has caused devastating declines in North American bat populations, there have been no apparent population changes attributable to the disease in Europe. We screened 276 bats of 15 species from hibernacula in the Czech Republic over 2012 and 2013, and provided histopathological evidence for 11 European species positive for WNS. With the exception of <i>Myotis myotis</i>, the other ten species are all new reports for WNS in Europe. Of these, <i>M. emarginatus, Eptesicus nilssonii, Rhinolophus hipposideros, Barbastella barbastellus</i> and <i>Plecotus auritus</i> are new to the list of <i>P. destructans</i>-infected bat species. While the infected species are all statistically phylogenetically related, WNS affects bats from two suborders. These are ecologically diverse and adopt a wide range of hibernating strategies. Occurrence of WNS in distantly related bat species with diverse ecology suggests that the pathogen may be a generalist and that all bats hibernating within the distribution range of <i>P. destructans</i> may be at risk of infection.</p></div

Neighbour-joining tree based on ecological and behavioural traits of bats from Europe and North America (rooted at midpoint).

<p>See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097224#pone-0097224-g002" target="_blank">Figure 2</a> for a description of the colour scheme.</p

Video_1_Case report: Filarial infection of a parti-coloured bat: Litomosa sp. adult worms in abdominal cavity and microfilariae in bat semen.MP4

BackgroundFilarial infections have been understudied in bats. Likewise, little is known about pathogens associated with the reproductive system in chiropterans. While semen quality is critical for reproductive success, semen-borne pathogens may contribute to reproductive failure.MethodsFor the first time we performed electroejaculation and used computer-assisted semen analysis to provide baseline data on semen quality in a parti-coloured bat (Vespertilio murinus).ResultsThe semen quality values measured in the V. murinus male appeared high (semen concentration = 305.4 × 106/mL; progressive and motile sperm = 46.58 and 60.27%, respectively). As an incidental finding, however, microfilariae were observed in the bat semen examined. At necropsy, eight adult filarial worms, later genetically identified as Litomosa sp., were found in the peritoneal cavity, close to the stomach, of the same particoloured bat male dying as a result of dysmicrobia and haemorrhagic gastroenteritis in a wildlife rescue centre. Histopathology revealed microfilariae in the testicular connective tissue and the epidydimal connective and fat tissues. A PCR assay targeting cytochrome c oxidase subunit 1 confirmed that adult worms from the peritoneal cavity and testicular microfilariae were of the same filarial species. Mildly engorged argasid mite larvae attached to the bat skin proved negative for filarial DNA and the adult filarial worms proved negative for endosymbiont Wolbachia.ConclusionWhile the standard filarial life cycle pattern involves a vertebrate definitive host and an invertebrate vector, represented by a blood-sucking ectoparasite, our finding suggests that microfilariae of this nematode species may also be semen-borne, with transmission intensity promoted by the polygynous mating system of vespertilionid bats in which an infected male mates with many females during the autumn swarming. Presence of microfilariae may be expected to decrease semen quality and transmission via this route may challenge the success of reproductive events in females after mating. Further investigation will be necessary to better understand the bat-parasite interaction and the life cycle of this filarial worm.</p

<i>Pseudogymnoascus destructans</i> infection in relation to chiropteran phylogeny and ecological similarity.

a<p> =  species using both types of shelters are also included in the previous categories.</p><p>Phylogenetic signal of explanatory variables on a phylogeny of bats from Europe and North America and of <i>P. destructans</i> infection on both phylogeny and a neighbour-joining tree based on squared Euclidean distances of ecological and behavioural traits of hibernating bat species. Values in bold indicate significant clustering or over-dispersion of <i>P. destructans</i> infection on the tree. Note that all categories of explanatory variables were tested here, but <i>n</i> - 1 dummy variables were included in the PGLS model. <i>N</i>  =  number of species scored positive for the given variable, MPD  =  mean phylogenetic distance, NRI  =  net relatedness index, MNTD  =  mean nearest taxon phylogenetic distance, NTI  =  nearest taxon index.</p

Histopathological skin lesions consistent with white-nose syndrome in ten European bat species.

<p>(A) <i>Myotis emarginatus</i>, (B) <i>Eptesicus nilssonii</i>, (C) <i>Rhinolophus hipposideros</i>, (D) <i>Plecotus auritus</i>, (E) <i>Barbastella barbastellus</i>, (F) <i>M. dasycneme</i>, (G) <i>M. nattereri</i>, (H) <i>M. daubentonii</i>, (I) <i>M. bechsteinii</i>, (J) <i>M. brandtii</i>. The photographs illustrate i) extensive infection of the wing membrane and cup-shaped epidermal erosions (A, E, H, J; long black arrow); ii) cup-like epidermal erosions in the pinna (B; long black arrow), iii) <i>Pseudogymnoascus destructans</i> hyphae obscuring the basement membrane and invading the dermis (A, B, C, E, H; black arrow); iv) a single cupping erosion packed with fungal hyphae in the wing membrane (C, D, G, I; long black arrow); v) colonisation of a hair follicle by <i>P. destructans</i>, fungal hyphae present in the associated sebaceous gland and regional connective tissue (F; black arrow); vi) marked signs of inflammation (B, F, J); and vii) a cellular inflammatory crust that sequesters fungal hyphae (A, J). White arrows within each photograph indicate the interface between epidermis and dermis. Periodic acid-Schiff stain; scale bar  =  50 µm. <i>M. myotis</i> not shown because WNS lesions in this species have already been documented elsewhere <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097224#pone.0097224-Pikula1" target="_blank">[18]</a>.</p

Boxplot of fitted values from the phylogenetic generalised least squares model of <i>P. destructans</i> infection.

<p>Predictions for infected and non-infected species overlap.</p