Chlamydia pecorum Ovine Abortion: Associations between Maternal Infection and Perinatal Mortality.

Chlamydia pecorum is a common gastrointestinal inhabitant of livestock but infections can manifest in a broad array of clinical presentations and in a range of host species. While C. pecorum is a known cause of ovine abortion, clinical cases have only recently been described in detail. Here, the prevalence and sequence types (STs) of C. pecorum in ewes from a property experiencing high levels of perinatal mortality (PNM) in New South Wales (NSW), Australia, were investigated using serological and molecular methods. Ewes that were PNM+ were statistically more likely to test seropositive compared to PNM- ewes and displayed higher antibody titres; however, an increase in chlamydial shedding from either the rectum, vagina or conjunctiva of PNM+ ewes was not observed. Multilocus sequence typing (MLST) indicated that C. pecorum ST23 was the major ST shed by ewes in the flock, was the only ST identified from the vaginal site, and was the same ST detected within aborted foetal tissues. Whole genome sequencing of C. pecorum isolated from one abortion case revealed that the C. pecorum plasmid (pCpec) contained a unique deletion in coding sequence 1 (CDS1) that was also present in C. pecorum ST23 shed from the ewes. A further unique deletion was noted in a polymorphic membrane protein gene (pmpG) of the C. pecorum chromosome, which warrants further investigation given the role of PmpG in host cell adherence and tissue tropism.This study describes novel infection parameters in a sheep flock experiencing C. pecorum-associated perinatal mortality, provides the first genomic data from an abortigenic C. pecorum strain, and raises questions about possible links between unique genetic features of this strain and C. pecorum abortion

A controlled antibiotic release system to prevent orthopedic-implant associated infections: An in vitro study

A new device for local delivery of antibiotics is presented, with potential use as a drug-eluting fixation pin for orthopedic applications. The implant consists of a stainless steel hollow tubular reservoir packed with the desired antibiotic. Release takes place through several orifices previously drilled in the reservoir wall, a process that does not compromise the mechanical properties required for the implant. Depending on the antibiotic chosen and the number of orifices, the release profile can be tailored from a rapid release of the load (ca. 20 h) to a combination of rapid initial release and slower, sustained release for a longer period of time (ca. 200 h). An excellent bactericidal action is obtained, with 4-log reductions achieved in as little as 2 h, and total bacterial eradication in 8 h using 6-pinholed implants filled with cefazolin

Biocompatibility Studies of Local Antibiotic-eluting Devices for Orthopaedic Applications

Introduction: Efficient local antibiotic-eluting devices could be an alternative for delivery of locally therapeutic antibiotics into tissues, avoiding bacterial contamination on implanted materials and minimizing side effects. A proper assessment of biocompatibility of the biomaterials used is important to improve safety after implantation. We present cytotoxicological and implantation tests results to evaluate biocompatibility of two drug-eluting systems with potential use in orthopaedic implants. Materials and Methods: Cytotoxicological studies were carried out by evaluating the in-vitro dose-dependent effect of cefazolin and linezolid on fibroblasts, keratinocytes, macrophages and osteoblasts. Cells were incubated with antibiotic concentrations ranging from 0.25 to 1.5 mg/ml. Cellular viability was assessed by the Alamar blue test. Cell cycle and apoptosis were measured by flow cytometry. Short-term implantation tests were performed in an ovine model to assess the local effects of the device. Two implants were used: (1) a macroporous stainless steel reservoir loaded with linezolid and (2) stainless steel pins with orifices drilled in the reservoir wall loaded with cefazolin. Implants were placed in sheep tibias. Tissues were studied by pathological means, determining the local effect and tissue response from the implant. (Ethical committee approval number: PI36/14) Results: Cytotoxic effects of cefazolin and linezolid were only found at 1.5 mg/ml on keratinocytes and osteoblasts, respectively. There were no significant changes on cell cycle and apoptosis at 1.0 mg/ml. Sheep with both antibiotic-loaded implants did not show local or systemic adverse effects. Conclusions: These results showed no potential toxic effects for the designed devices. However, the antibiotic local concentration should not exceed 1.0 mg/ml

Case report : synucleinopathy associated with phalaris neurotoxicity in sheep

Chronic intoxication with tryptamine-alkaloid-rich Phalaris species (spp.) pasture plants is known colloquially as Phalaris staggers syndrome, a widely occurring neurological disorder of sheep, cattle, horses, and kangaroos. Of comparative interest, structurally analogous tryptamine-alkaloids cause experimental parkinsonism in primates. This study aimed to investigate the neuropathological changes associated with spontaneous cases of Phalaris staggers in sheep with respect to those encountered in human synucleinopathy. In sheep affected with Phalaris staggers, histological, immunohistochemical, and immunofluorescence analysis revealed significant accumulation of neuromelanin and aggregated α-synuclein in the perikaryon of neurons in the cerebral cortex, thalamus, brainstem, and spinal cord. Neuronal intracytoplasmic Lewy bodies inclusions were not observed in these cases of ovine Phalaris staggers. These important findings established a clear link between synucleinopathy and the neurologic form of Phalaris plant poisoning in sheep, demonstrated in six of six affected sheep. Synucleinopathy is a feature of a number of progressive and fatal neurodegenerative disorders of man and may be a common endpoint of such disorders, which in a variety of ways perturb neuronal function. However, whether primary to the degenerative process or a consequence of it awaits clarification in an appropriate model system

A dilated pore of winer in a young Iberian lynx (Lynx Pardinus)

A 1.5-year-old male Iberian lynx born in captivity presented with a 2.2 cm exophytic nodular alopecic mass at the left tarsus medial aspect, before being released into the wild. The lesion had keratinised material protruding through a 1.2 cm pore, forming a cutaneous horn. Histopathology revealed a cystic dermo-epidermal lesion lined by psoriasiform hyperplasia of the epithelium, thicker at the base of the cyst, together with abrupt to progressive keratinization and keratin pearls. The central cystic cavity was filled with fibrillary keratin and ghost cells. Immunohistochemistry for pan-cytokeratin showed strong positive immunolabeling in the hyperplastic epithelium and a weaker immunolabeling at the content of the cyst. A final diagnosis of a Dilated Pore of Winer was made. This paper is the first reported case of a DPW in a lynx and a wild felid

Epithelial–Mesenchymal Transition in a Case of Metastatic Thyroid Carcinoma in a Brown Bear (Ursus arctos)

A 20-year-old male brown bear (Ursus arctos) with a 20 × 25 cm necrotic mass adjacent to the trachea was diagnosed as having an anaplastic thyroid carcinoma. Metastases were observed in the lungs and one adrenal gland and, histologically, these had anaplastic and follicular carcinoma patterns, respectively. E-cadherin labelling was observed in the adrenal mass only, while N-cadherin immunolabelling was detected in the thyroid gland and lung masses. Thyroid-specific markers (thyroid transcription factor-1, thyroglobulin) were expressed in the adrenal gland metastasis. This case illustrates an example of a primary epithelial–mesenchymal transition (EMT) enabling metastasis to distant organ sites, followed by a mesenchymal–epithelial transition within the adrenal gland microenvironment, allowing invasion and reacquisition of thyroid epithelial cell features. EMTs help to understand the phenomenon of carcinoma cell plasticity in enabling colonization and growth of metastases

On the improvement of alveolar-like microfluidic devices for efficient blood oxygenation

In this work, we study alveolar-like microfluidic devices with a horizontal membrane arrangement that demonstrate a great potential as small-scale blood oxygenator. The design criteria for the fabricated devices were to maximize the oxygen saturation level and minimize liquid chamber volume while ensuring the physiological blood flow in order to avoid thrombus formation and channel blockage during operation. The liquid chamber architecture was iteratively modified upon analysis of the fluid dynamics by computer modelling. Accordingly, two alveolar type architectures were fabricated, Alveolar Design 1 (AD1) and Alveolar Design 2 (AD2), and evaluated for oxygenation of sheep blood. The attained O2 transfer rate at 1 mL/min of blood flow rate for both devices was rather similar: 123 mL·min-1 ·m-2 and 127 mL·min-1 ·m-2 for AD1 and AD2 microfluidic devices, respectively. Among the studied, AD2 type geometry would lead to the lowest pressure drop and shear stress value upon implementation in a scaled microfluidic artificial lung (µAL) to satisfy oxygenation requirements of a 2.0 kg neonate.Government of Aragon and the Education, Audiovisual and Culture Executive Agency (EU-EACEA) within the EUDIME – ‘Erasmus Mundus Doctorate in Membrane Engineering’ program (FPA 2011-0014, SGA 2012-1719, http://eudime.unical.it). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008–2011 financed by the Instituto de Salud Carlos III with the assistance of the European Regional Development Fund

Diagnosing infection with small ruminant lentiviruses of genotypes A and B by combining synthetic peptides in ELISA

The major challenges in diagnosing small ruminant lentivirus (SRLV) infection include early detection and genotyping of strains of epidemiological interest. A longitudinal study was carried out in Rasa Aragonesa sheep experimentally infected with viral strains of genotypes A or B from Spanish neurological and arthritic SRLV outbreaks, respectively. Sera were tested with two commercial ELISAs, three based on specific peptides and a novel combined peptide ELISA. Three different PCR assays were used to further assess infection status.The kinetics of anti-viral antibody responses were variable, with early diagnosis dependent on the type of ELISA used. Peptide epitopes of SRLV genotypes A and B combined in the same ELISA well enhanced the overall detection rate, whereas single peptides were useful for genotyping the infecting strain (A vs. B). The results of the study suggest that a combined peptide ELISA can be used for serological diagnosis of SRLV infection, with single peptide ELISAs useful for subsequent serotyping.Funded by CICYT (AGL2010-22341-C04-01 and AGL2013-49137-C3-1R) and Navarra's Government (IIQ010449.RI1 and IIQ14064.RI1). L. Sanjosé was a FPI-fellow of the Spanish MINECO and R. Reina had a contract of the Public University of Navarra.Peer Reviewe

Cognition and behavior in sheep repetitively inoculated with aluminum adjuvant-containing vaccines or aluminum adjuvant only

Sheep health management strategies often include the use of aluminum (Al)-containing vaccines. These products were associated with the appearance of the ovine autoimmune/inflammatory syndrome induced by adjuvants (ASIA syndrome), which included an array of ethological changes in the affected animals. The aim of this pilot study was to investigate cognitive and behavioral changes in sheep subjected to a protocol of repetitive inoculation with Al-containing products. Twenty-one lambs were assigned to three groups (n = 7 each): Control, Adjuvant-only, and Vaccine. Vaccine group was inoculated with commercial Al- hydroxide containing vaccines; Adjuvant-only group received the equivalent dose of Al only (Alhydrogel®), and Control group received Phosphate-buffered saline. Sixteen inoculations were administered within a 349-day period. Ethological changes were studied in late summer (7 inoculations) and mid-winter (16 inoculations). Animals in Vaccine and Adjuvant-only groups exhibited individual and social behavioral changes. Affiliative interactions were significantly reduced, and aggressive interactions and stereotypies increased significantly. They also exhibited a significant increase in excitatory behavior and compulsive eating. There were increased levels of stress biomarkers in these two groups. In general, changes were more pronounced in the Vaccine group than they were in the Adjuvant-only group. Some changes were already significant in summer, after seven inoculations only. This study is the first to describe behavioral changes in sheep after having received repetitive injections of Al-containing products, and may explain some of the clinical signs observed in ovine ASIA syndrome

Growth performance and clinicopathological analyses in lambs repetitively inoculated with aluminum-hydroxide containing vaccines or aluminum-hydroxide only

Aluminum (Al) hydroxide is an effective adjuvant used in sheep vaccines. However, Al-adjuvants have been implicated as potential contributors to a severe wasting syndrome in sheep— the so-called ovine autoimmune-inflammatory syndrome induced by adjuvants (ASIA syndrome). This work aimed to characterize the effects of the repetitive injection of Al-hydroxide containing products in lambs. Four flocks (Flocks 1–4; n = 21 each) kept under different conditions were studied. Three groups of seven lambs (Vaccine, Adjuvant-only, and Control) were established in each flock. Mild differences in average daily gain and fattening index were observed, indicating a reduced growth performance in Vaccine groups, likely related to short-term episodes of pyrexia and decreased daily intake. Clinical and hematological parameters remained within normal limits. Histology showed no significant differences between groups, although there was a tendency to present a higher frequency of hyperchromatic, shrunken neurons in the lumbar spinal cord in the Adjuvant-only group. Although Al-hydroxide was linked to granulomas at the injection site and behavioral changes in sheep, the results of the present experimental work indicate that injected Al-hydroxide is not enough to fully reproduce the wasting presentation of the ASIA syndrome. Other factors such as sex, breed, age, production system, diet or climate conditions could play a role