Dispersal distances for airborne spores based on deposition rates and stochastic modeling

ABSTRACT A new modeling framework for particle dispersal is explored in the context of the particles being fungal spores dispersed within a field. The model gives rise to both exponentially decreasing and polynomially decreasing two-dimensional densities of deposited fungal spores. We reformulate the model in terms of time to deposition, and show how this concept is equivalent to the deposition rate for fungal spores. Special cases where parameter values for wind and gravitation lead to exponentially or polynomially decreasing densities are discussed, and formulas for one- and two-dimensional densities of deposited spores are given explicitly in terms of parameters for diffusion, wind, gravitation, and spore release height.     

Streptococcus equi subsp. zooepidemicus isolates from equine infectious endometritis belong to a distinct genetic group

Streptococcus equi subsp. zooepidemicus is the pathogen most commonly isolated from the uterus of mares. S. zooepidemicus is an opportunistic pathogen and part of the resident flora in the caudal reproductive tract. The aim of this study was to investigate whether a genotypically distinct subpopulation of S. zooepidemicus is associated with endometritis in the mare, by genotyping and comparing uterine S. zooepidemicus strains with isolates from the vagina and clitoral fossa. Mares with (n = 18) or without (n = 11) clinical symptoms of endometritis were included. Uterine samples were obtained using a guarded endometrial biopsy punch, whereas a swab was used to recover samples from the cranial vagina and the clitoral fossa. If S. zooepidemicus was present, up to three colonies were selected from each anatomical location (max. 9 isolates per mare). Bacterial isolates were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). S. zooepidemicus was isolated from the endometrium of 12 mares. A total of 88 isolates were analyzed by PFGE: 31 from the endometrium, 26 from the cranial vagina and 31 isolates from the clitoral fossa. For MLST 21 isolates were chosen. Results demonstrated a higher genetic similarity of the isolates obtained from infectious endometritis compared to isolates obtained from the caudal reproductive tract. In conclusion, we demonstrate for the first time that a genetically distinct group of S. zooepidemicus is associated with infectious endometritis in the mare.     

The rarely reported tet(31) tetracycline resistance determinant is common in Gallibacterium anatis

The present investigation was undertaken to identify and characterize the tetracycline resistance determinant in 22 Gallibacterium anatis strains for which no determinant was identified using primers specific for tet(A, B, C, D, E, G, H, K, L, M, O). A recent study found tet(B) to be the most prevalent tetracycline resistance determinant in a larger collection of G. anatis field strains from Mexico and Denmark. However, in 41% of the tetracycline resistant strains no determinant could be assigned. Here we demonstrate that tet(31) is a common determinant in G. anatis originating from chickens from very different production systems and localities. In addition, tet(31) was identified in strains isolated over a 30-year period. This is the first report on tet(31) since its original identification in Aeromonas salmonicida.     

A non-radiometric method for measuring serum thymidine kinase activity in malignant lymphoma in dogs

The aim of this study was to evaluate an enzyme-linked immunosorbent assay (ELISA), for determination of serum thymidine kinase 1 (sTK1) activity in dogs with malignant lymphoma (ML) and compare it with a thymidine kinase (TK) radioenzymatic assay (TK-REA). The TK-REA has recently been shown to be useful in determining the clinical stage and prognosis in canine ML. In addition, serum lactate dehydrogenase (LDH) was measured. Forty-five dogs were included in the study. Sixty serum samples from these dogs, stored in a tumour serum sample bank (stored at -20 degrees C), were analysed. Apart from 37 dogs with ML, four normal dogs as well as two dogs with mammary carcinomas, one dog with bladder carcinoma, and one dog with malignant fibrous histiocytoma were included. Staging of ML was based on the modified World Health Organization (WHO) staging system for canine ML. The diagnosis of all tumours was verified by histopathology. The TK activity (units per litre [U/L]) ranged from 1.0 to 607.9 in the TK-REA analysis and from 1.1 to 510 in the TK-ELISA (normal reference value <7U/L). The range for LDH was between 12 and 1194 U/L (normal reference value <228 U/L). There was a significant correlation between the TK-REA and the TK-ELISA. The correlation coefficient (CC) was 0.97 and the standard error of the estimate (SEE) was 3.7 U/L. There was no correlation between LDH and either the TK-REA or the TK-ELISA (CC=0.53 for both assays; SEE=26.7 and 12.7 U/L, respectively). Most of the variation in LDH was still within the normal reference range. The mean LDH in dogs with high-stage (stage IV+V) disease was 201.9 U/L. The corresponding values for the TK-REA and TK-ELISA were 109 and 109.9 U/L, respectively. The significant relation between the TK-REA and the TK-ELISA was confirmed by Bland-Altman analysis. The TK-ELISA assay, because of its relative simplicity, will permit measurement of TK in cases of ML in dogs to become a routine procedure.     

In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

Metabolic characterization of rumen epithelial tissue from dairy calves fed different starter diets using 1H NMR spectroscopy

The metabolic profile of calf rumen epithelial tissue was for the first time characterized by ¹H NMR spectroscopy of chloroform/methanol extracts. The metabolite profiles comprised a number of amino acids, creatine, taurine, short-chain fatty acids and triglycerides. The effects of two dietary interventions; i) four levels of milk allowance with a concomitant different uptake of starter concentrate, and ii) two diets with varying content of starch and fibre in the concentrate, were elucidated. Partial least square regression analysis revealed that the intensity of NMR signals assigned to leucine, isoleucine and valine (0.90  ppm), propionate (1.06 and 2.18  ppm), lactate (1.32  ppm), butyrate (1.56  ppm), acetate (1.93  ppm), glutamine/glutamate (2.18, 2.35 and 3.75  ppm), creatine (3.04 and 3.94  ppm) and glycine (3.55  ppm) decreased with increasing milk allowance. In addition, the analysis revealed that the main difference between the two diets was the content of propionate in the epithelia tissue extracts. Previous morphological analyses of the same rumen epithelia were not able to detect any significant effects of either milk allowance or dietary starch content. Accordingly, the present study demonstrated that ¹H NMR spectroscopy applied on extracts is a useful tool for metabolite profiling of epithelial tissue and for following the development of epithelia tissue in young calves, and that the technique may be more sensitive to dietary effects than morphological studies.     

Immunochemical analyses of serum antibodies from pig herds in a Salmonella non-endemic region.

In a large comparative survey of Danish and Swedish slaughter pig herds performed prior to this work, it was unexpectedly found that some Swedish herds harbored seropositive pigs. Serum samples from the Swedish herds had moderate responses in the Salmonella mix-ELISA (detecting serogroup B and C1 infections) compared to the Danish herds classifying some of them as seropositive using a cut-off value at 40 OD%. In Sweden, extensive Salmonella control is carried out by bacteriological screening of feces and lymph nodes, and the overall prevalence has been proven to be below 0.1%. The serological positive results were therefore unexpected; hence the reactivities of the Swedish sera were studied by a number of immunochemical analyses (Western blot, indirect ELISA, inhibition ELISA, avidity ELISA) and compared to sera from Danish pig herds with verified Salmonella infections ("the reference sera").In Western blot, the Swedish sera had high binding reactivities against Salmonella Typhimurium LPS of different molecular weights, and gave binding patterns similar to that of the reference sera. Pre-incubation with free S. Typhimurium LPS or PS (the polysaccharide part of LPS) was able to inhibit the reactivity of the Swedish sera in the mix-ELISA. Reactivities against other related bacterial LPS such as Citrobacter freundii LPS and Yersinia enterocolitica O:3 LPS were observed in the Swedish sera, but these LPS antigens were unable to inhibit the reactivities in the mix-ELISA as efficiently as S. Typhimurium LPS. Furthermore, the Swedish sera did not bind Salmonella LPS of another serogroup (S. Meleagridis LPS, serogroup E1) or rough Salmonella LPS, both lacking the specific O-antigenic parts of S. Typhimurium LPS. The avidity of the Swedish sera was much lower than the avidity of the reference sera, which could indicate the presence of transient low-dose infections or stimulation by inactivated bacteria in feed. The results obtained in this investigation strongly indicate that the Swedish sera contain antibodies directed against the O-antigenic part of LPS from S. Typhimurium or possibly on as yet unknown bacterium.     

Genetic relatedness and netB prevalence among environmental Clostridium perfringens strains associated with a broiler flock affected by mild necrotic enteritis

In a previous study we investigated pulsed-field gel electrophoresis (PFGE) genotype diversity and prevalence of the netB toxin gene in Clostridium perfringens (CP) isolates recovered from a broiler flock (flock 1) affected by necrotic enteritis (NE). In this follow-up work, we examined samples collected before placement of flock 1, to see if NE during rearing could be traced back to the cleaned and empty building or the day-old chicks. Litter from the next flock in the same building (flock 2) was also examined. We detected 25 different PFGE genotypes, five of which were found only in litter from flock 2. Six genotypes which had been found in flock 1 during rearing were detected in samples collected before placement. NetB positive isolates belonging to two of these genotypes had been recovered from NE lesions during rearing, suggesting that virulent strains were transmitted from the cleaned and disinfected broiler house. NetB frequency among isolates from the empty building was 45%, indicating that netB positive strains were prevalent in a building that previously had housed a healthy flock offered in-feed narasin (flock 0). NetB frequency among isolates from litter used by flock 2 was 22%, indicating that netB positive strains were present in the environment of a 14-days-old healthy flock offered in-feed narasin. Two prevalent genotypes were consistently either netB negative or netB positive. However, the presence of genotypes represented by both negative and positive isolates may suggest that the gene can spread horizontally among different CP strains.     

Single Layer Centrifugation of Stallion Spermatozoa through Androcoll™‐E does not Adversely Affect their Capacitation‐Like Status,as Measured by CTC Staining

This study was designed to evaluate the effect of single layer centrifugation (SLC) and subsequent cold storage on stallion sperm capacitation‐like status and acrosome reaction. Three stallions were included in the study, with three ejaculates per stallion. The samples were examined 4, 24 and 72 h after collection, extension and SLC, with storage at 6°C. Sperm capacitation‐like status was investigated using the fluorescent dye chlortetracycline (CTC). There was no difference in capacitation‐like status between colloid‐selected and non‐selected spermatozoa. Sperm motility decreased significantly during cold storage, whereas the proportion of apparently capacitated spermatozoa increased. There was no change in the proportion of acrosome‐reacted spermatozoa. In conclusion, SLC through Androcoll?‐E does not adversely affect the capacitation‐like status of stallion spermatozoa, although it did increase with time during cold storage.