Kinetics of local and systemic leucocyte and cytokine reaction of calves to intrabronchial infection with Chlamydia psittaci

PLOS One; Volume 10, Band 8, p.1-21   https://doi.org/10.1371/journal.pone.0135161

(Annette Prohl, Katharina Wolf, Corinna Weber, Elisabeth Müller, Christian Menge, Konrad Sachse, Juergen Roedel, Petra Reinhold, Angela Berndt)

Abstract

Infection of cattle with chlamydiae is ubiquitous and, even in the absence of clinical sequeleae, has a quantifiable negative impact on livestock productivity. Despite recent progress, our knowledge about immune response mechanisms capable of counteracting the infection and preventing its detrimental effects is still limited. A well-established model of bovine acute respiratory Chlamydia (C.) psittaci infection was used here to characterize the kinetics of the local and systemic immune reactions in calves. In the course of two weeks following inoculation, leukocyte surface marker expression was monitored by flow cytometry in blood and bronchoalveolar lavage fluid (BALF). Immune-related protein and receptor transcription were determined by quantitative real-time reverse transcription PCR in blood, BALF and lung tissue. An early increase of IL2RA, IL10 and HSPA1A mRNA expressions was followed by a rise of lymphocytes, monocytes, and granulocytes exhibiting activated phenotypes in blood. Monocytes showed elevated expression rates of CD11b, CD14 and MHC class II. The rates of CD62L expression on CD8hi T cells in blood and on CD4+ T cells in BALF were also augmented and peaked between 2 and 4 dpi. Notably, CD25 antigen expression was significantly elevated, not only on CD8dim/CD62L+ and CD8-/CD62L+ cells in blood, but also on granulocytes in blood and BALF between 2–3 dpi. From 4 dpi onwards, changes declined and the calves recovered from the infection until 10 dpi. The findings highlight the effectiveness of rapid local and systemic immune reaction and indicate activated T cells, monocytes and granulocytes being essential for rapid eradication of the C. psittaci infection.

Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle

Karanikola et al. Parasites & Vectors (2015) 8:335 

(Sofia N. Karanikola, Jürgen Krücken, Sabrina Ramünke, Theo de Waal, Johan Höglund, Johannes Charlier, Corinna Weber, Elisabeth Müller, Slawomir J.Kowalczyk, Jaroslaw Kaba, Georg von Samson-Himmelstjerna and Janina Demeler)

Abstract

A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. The capability of a multiplex platform for the simultaneous detection of three pasture associated parasite species was evaluated and compared to existing ELISAs. The recombinant antigens 14.2 kDa ES protein for Cooperia oncophora, major sperm protein for Dictyocaulus viviparus and Cathepsin L1 for Fasciola hepatica were recombinantly expressed either in Escherichia coli or Pichia pastoris. Antigens were covalently coupled onto magnetic beads. Optimal concentrations for coupling were determined following the examination of serum samples collected from experimentally mono-infected animals, before and after their infection with the target species. Absence of cross-reactivity was further determined with sera from calves mono-infected with Haemonchus contortus, Ostertagia ostertagi and Trichostrongylus colubriformis. Examination of negative serum samples was characterised by low median fluorescence intensity (MFI). Establishment of the optimal serum dilution of 1:200 was achieved for all three bead sets. Receiver Operating Characteristic analyses were performed to obtain cut-off MFI values for each parasite separately. Sensitivity and specificity at the chosen cut-off values were close to, or 100 % for all bead sets. Examination of serum samples collected on different days post infection from different animals showed a high reproducibility of the assays. Serum samples were additionally examined with two already established ELISAs, an in-house ELISA using the recombinant MSP as an antigen and a DRG ELISA using Cathepsin L1 for liver fluke. The results between the assays were compared and kappa tests revealed an overall good agreement. A versatile bead-based assay using fluorescence detection (xMAP® technology) was developed to simultaneously detect antibodies against C. oncophora, D. viviparus and F. hepatica in cattle serum samples. This platform provides rapid, high-throughput results and is highly sensitive and specific in comparison to existing serological as well as coproscopical diagnostic techniques.

Detection of feline coronavirus spike gene mutations as a tool to diagnose feline infectious peritonitis

(Felten S, Weider K, Doenges S, Gruendl S, Matiasek K, Hermanns W, Mueller E, Matiasek L, Fischer A, Weber K, Hirschberger J, Wess G, Hartmann K.)

Abstract

OBJECTIVES: Feline infectious peritonitis (FIP) is an important cause of death in the cat population worldwide. The ante-mortem diagnosis of FIP in clinical cases is still challenging. In cats without effusion, a definitive diagnosis can only be achieved post mortem or with invasive methods. The aim of this study was to evaluate the use of a combined reverse transcriptase nested polymerase chain reaction (RT-nPCR) and sequencing approach in the diagnosis of FIP, detecting mutations at two different nucleotide positions within the spike (S) gene.

METHODS: The study population consisted of 64 cats with confirmed FIP and 63 cats in which FIP was initially suspected due to similar clinical or laboratory signs, but that were definitively diagnosed with another disease. Serum/plasma and/or effusion samples of these cats were examined for feline coronavirus (FCoV) RNA by RT-nPCR and, if positive, PCR products were sequenced for nucleotide transitions within the S gene.

RESULTS: Specificity of RT-nPCR was 100% in all materials (95% confidence interval [CI] in serum/plasma 83.9-100.0; 95% CI in effusion 93.0-100.0). The specificity of the sequencing step could not be determined as none of the cats of the control group tested positive for FCoV RNA. Sensitivity of the 'combined RT-nPCR and sequencing approach' was 6.5% (95% CI 0.8-21.4) in serum/plasma and 65.3% (95% CI 50.4-78.3) in effusion.

CONCLUSIONS AND RELEVANCE: A positive result is highly indicative of the presence of FIP, but as none of the control cats tested positive by RT-nPCR, it was not possible to confirm that the FCoV mutant described can only be found in cats with FIP. Further studies are necessary to evaluate the usefulness of the sequencing step including FCoV-RNA-positive cats with and without FIP. A negative result cannot be used to exclude the disease, especially not when only serum/plasma samples are available.

Herd-level seroprevalence of Neospora caninuminfectionin dairy cattle in central and northeastern Poland

W. Stefański Institute of Parasitology, PAS   DOI: 10.1515/ap-2016-0006

Acta Parasitologica, 2016, 61(1), 63–65; ISSN 1230-2821

(Sławomir J. Kowalczyk, Michał Czopowicz, Corinna N. Weber, Elisabeth Müller, Lucjan Witkowski and Jarosław Kaba)

Abstract

A serosurvey was carried out to estimate the herd-level seroprevalence of Neospora caninuminfection in cattle in central andnortheasternPoland. Ninety seven dairy cattle herds from 2 provinces of Poland (Podlaskie, 47 herds and Łódzkie, 50 herds)were randomly enrolled in the study using two-stage cluster method. A simple random selection was applied within each herdto select a sample of adult cows (≥18 month-old). A total number of 734 cows were enrolled in the study. The animals werescreened with a commercial competitive ELISA (Bio-X Diagnostics, Belgium). To calculate true herd-level seroprevalence testsensitivity and specificity were adjusted from an individual- to a herd-level using FreeCalc method. The true overall herd-levelseroprevalence of N. caninuminfection was 56.7% (95% CI: 47.5%, 65.9%). The true herd-level seroprevalence in Podlaskiewas 63.3% (95% CI: 43.0%, 83.6%) and 50.5% (95% CI: 32.8%, 68.2%) in Łódzkie province and these figures did not differsignificantly between the two provinces (chi2test p = 0.238). One hundred forty three of 734 cows (19.5%) were seropositivewhich gave the true overall individual-level seroprevalence of 20.1% (95% CI: 17.4%, 23.2%). Percentage of seropositivecows in each herd varied from 6% to 80%. This study is the first epidemiological investigation of herd-level seroprevalence ofN. caninuminfection in Polish dairy cattle population. In conclusion, the result of the study confirmed previous data that N. can-inuminfection is widespread in the Polish cattle population and thus should be considered as a potential cause of spontaneousabortions.

Encephalitozoon cuniculi causes focal anterior cataract and uveitis in dogs

Journal: Tierärztliche Praxis Kleintiere, Heft 5 2015 ; p.337-344

dx.doi.org/10.15654/TPK-141053

(B. Nell, J. Csokai, A. Fuchs-Baumgartinger, G. Maaß)

Abstract

Three mongrel dogs, aged 10 months (case 1), 14 months (case 2) and 7.5 years (case 3), were presented because of ophthalmologic disorders of 4 months, 6 months and 7 years duration, respectively. All three dogs were offspring of stray dogs from Hungary and Serbia and had positive serum antibody titres against Encephalitozoon (E.) cuniculi. The two young dogs showed unilateral, the older dog bilateral chronic anterior uveitis with posterior synechia and focal anterior cortical cataract. The fundi that could be evaluated developed focal tapetal hyporeflective lesions in the course of the disease. Dogs 1 and 2 underwent removal of the lens via phacoemulsification. PCR of the lens material was positive for E. cuniculi strains IV and II, respectively. In dog 2 findings suggestive of microsporidia were detected underneath the anterior lens capsule by immunohistochemical staining. In all cases medical treatment consisted of systemic fenbendazole, prednisolone, and topical anti-inflammatory drugs, and additional brinzolamid/timolol for dog 3. For the time being all cases (follow up 23 months, 6 months and 3 months, respectively) are still on topical anti-inflammatory therapy. It is concluded that E. cuniculi infections can cause cataract and chorioretinal lesions in dogs.

Identification of snake arenaviruses in live boas and pythons in a zoo in Germany

(T. Aqrawi; A. C. Stöhr; T. Knauf-Witzens; A. Krengel; K. O. Heckers; R. E. Marschang)

Tierärztliche Praxis Kleintiere, (4-2015) Volume 43

dx.doi.org/10.15654/TPK-140743

Summary

Objective: Recent studies have described the detection and characterisation of new, snake specific arenaviruses in boas and pythons with inclusion body disease (IBD). The objective of this study was to detect arenaviral RNA in live snakes and to determine if these were associated with IBD in all cases. Samples for arenavirus detection in live aniated with IBD in all cases. Samples for arenavirus detection in live animals were compared. Detected viruses were compared in order to understand their genetic variability.

PDF Download >>

Detection of Bacteria in Oral Swabs from Healthy Common Musk Turtles (Sternotherus odoratus) and West African Mud Turtles (Pelusios castaneus)

Journal of Herpetological Medicine and Surgery Volume 25, No. 1–2, 2015; P.33-39

(Verena Heynol, Kim O. Heckers, Helge Behncke, Anton Heusinger, Rachel E. Marschang)

Abstract

The bacterial flora of clinically healthy reptiles can be extremely diverse. The objectives of this study were to determine the bacterial flora in oral swabs from two different species of water turtles and to study growth characteristics and antibiotic resistance of these isolates. Oral swabs were collected from 20 clinically healthy common musk turtles (Sternotherus odoratus) and 20 West African mud turtles (Pelusios castaneus) and incubated at two different temperatures (25 and 37°C; 77.0 and 98.6°F) on various agar plates. All isolates were tested for susceptibility to a panel of antibiotics. A total of 66 distinct bacterial types were collected, 63 (95.45%, 95% confidence interval [CI]: 90–100) of which were Gram negative and 3 (4.55%, 95% CI: 0–27) of which were Gram positive. The most commonly isolated genera were Citrobacter spp. (in 97.5% of the animals tested, 95% CI: 93–100), Aeromonas spp. (in 92.5% of the animals tested, 95% CI: 84–100), Chryseobacterium spp. (in 80% of the animals tested, 95% CI: 68–92), and Salmonella spp. (in 80% of the animals tested, 95% CI: 68–92). The most commonly isolated bacterium was Aeromonas hydrophila (77.5%, 95% CI: 65–90). The oral bacterial flora of all of the examined animals consisted of a wide mixture of bacteria, many of which were potential pathogens. The combination of bacteria detected differed between individual animals. Incubation at 25 and 37°C led to the detection of distinct populations of bacteria. The resistance testing showed that many of the bacteria detected were resistant to a wide range of antibiotics.

Sequencing and phylogenetic analysis identifies candidate members of a new picornavirus genus in terrestrial tortoise species

Arch Virol. 160, 2015: Springer-Verlag Wien 811-816

(Farkas SL, Ihász K, Fehér E, Bartha D, Jakab F, Gál J, Bányai K, Marschang RE)

Abstract

Near-complete genome sequences of seven picornavirus (PV) strains isolated from different terrestrial tortoise species were determined and characterized. The genome organization of the strains proved to be similar and displayed a typical PV layout, and the polyprotein-encoding regions showed low similarity to those of other PVs. The predicted regions of the tortoise PV genomes were related to the corresponding genome parts of viruses belonging to distinct genera, implying modular evolution of these novel viruses. Our results suggest that these tortoise PVs belong to a prototype species in a separate proposed genus in the family Picornaviridae, tentatively called Topivirus (Tortoise picornavirus).

Detection of a new alphaherpesvirus in West African mud turtles (Pelusios castaneus)

Proceedings 2nd ICARE Paris, France, 18.-23. April 2015.

(Marschang R, Heckers K, Heynol V, Weider K, Behncke H.)

Complete sequencing of ranavirsues detected in European reptiles

Proceedings 2nd ICARE Paris, France, 18.-23. April 2015.

(Stöhr AC, López-Bueno A, Blahak S, Herberg AA, Marschang RE.)

Formal pathogenesis and virus distribution after experimental infection of corn snakes with a ferlavirus

Proceedings 2nd ICARE Paris, France, 18.-23. April 2015.

(Pees M, Neul A, Plenz B, Truyen U, Leinecker N, Marschang RE, Starck M, Müller K, Schmidt V)

Detection of arenaviruses in live snakes

Farkas Proceedings 2nd ICARE Paris, France, 18.-23. April 2015. 

(Marschang RE, Kolesnik E, Aqrawi T, Diez J, Heckers KO)

Distribution and Host Range of Ranaviruses

Gray MJ, Chinchar VG (eds.). Ranaviruses: Lethal Pathogens of Ectothermic Vertebrates. 

SpringerOpen. DOI 10.1007/978-3-319-13755-1. Pp. 9-58.

(Duffus ALJ, Waltzek TB, Stöhr AC, Allender MC, Gotesman M, Whittington RJ, Hick P, Hines MK, Marschang RE)

Greek tortoises – Testudos don’t need tob e all Greek to you

Proceedings of the NAVC Conference Volume 29,

Orlando, Florida, USA, 17.-21. January 2015. 

(Marschang RE)

Why the dead crickets matter – invertebrate iridoviruses in reptiles and amphibians

Proceedings of the NAVC Conference Volume 29,

Orlando, Florida, USA, 17.-21. January 2015. 

(Marschang RE)

The 411 on inclusion body disease – what you need to know

Proceedings of the NAVC Conference Volume 29,

Orlando, Florida, USA, 17.-21. January 2015. 

(Marschang RE)

Reptile Virology – understanding it and how to make it profitable for your practice

Proceedings of the NAVC Conference Volume 29,

Orlando, Florida, USA, 17.-21. January 2015. 

(Wellehan JFX, Marschang RE)

Phyologeny and differentiation of reptilian and amphibian ranaviruses detected in Europe

(Stöhr AC, López-Bueno A, Blahak S, Caeiro MF, Rosa GM, Alves de Matos AP, Martel A, Alejo A, Marschang RE.)

PLOS ONE 10(2) 2015: e0118633. doi:10.1371/journal.pone.0118633