Host-cell apoptosis in Taenia solium-induced brain granulomas in naturally infected pigs

To assess whether apoptosis occurs in pig brain granulomas due to Taenia solium cysticerci, brain tissues from 30 pigs naturally infected with T. solium cysticercosis were evaluated by terminal deoxynucleotidyl transferase-end labelling (TUNEL) staining. In addition, tissues were stained with CD3 marker to identify T lymphocytes. Examination of TUNEL-stained tissues showed apoptotic cells in early lesions that contained viable cysticerci. Apoptotic cells were primarily found interspersed with normal cell types, and were mostly located in the inflammatory infiltrate. Late or advanced granulomas with disintegrated scolices did not show TUNEL-positive cells. CD3+ cells were found in both early and advanced lesions and apoptosis mainly co-localized with CD3+ T lymphocytes. This suggests that these cells are constantly undergoing apoptosis and thus die as soon as they arrive at the site of infection. Apoptosis indeed may be one way by which T. solium cysticerci down-regulate the host's cellular immune response in early cysticercosis. Therefore, further research is needed to establish if other cells besides T-lymphocytes are also a target for destruction by cysticerci in early cysticercosis as well as studies to assess if cysteine protease is expressed by viable cysticerci in situ.     

Improving hydration in elite male footballers during a national team training camp – an observational case study

[Purpose] The purpose of this study was to (i) assess hydration levels in elite male football players during a national team training camp before and during qualifying matches, (ii) evaluate the effect of coaching strategies for hydration based on feedback from hydration monitoring, and (iii) assess possible relationships between hydration status and training load or wellness markers.[Methods] Thirty-one male players (age 27±4 yrs; height 185±6 cm; weight 82.9±6.7 kg; body fat 10.4±2.3%) representing a national team from the Union of European Football Associations (UEFA) participated. The players were studied during three different national team training camps related to the UEFA Nations League tournament. Urine specific gravity (U_SG) was measured to assess hydration status. During all camps, the players were actively coached on improving strategies for hydration and given individual feedback on their test results. The training load was measured using GPS technology, and wellness questionnaires were completed.[Results] U_SG decreased progressively and significantly (p<0.005) during camp 1 and hydration status improved over the three camps, with fewer dehydrated and more well-hydrated players identified during the last part of camp 3. Significantly (p<0.05) higher U_SG values were observed 2 days prior to a match (MD-2) than on match day (MD); consequently, 52% of the players were dehydrated on MD-2 and only 6% on MD. No correlations were observed between hydration status and training load or wellness markers.[Conclusion] Dehydration is a challenge in elite male football, but continuous monitoring of hydration status and coaching on hydration strategies can lead to major improvements and reduce the degree of dehydration.     

Long‐term tag retention on two species of small cetaceans

The effects of tagging on small cetaceans are difficult to assess due to logistical difficulties in recapturing the whales. In this study two narwhals, Monodon monoceros, and five harbor porpoises, Phocoena phocoena, were recaptured between 297 and 767 days after instrumentation with satellite transmitters. The transmitters were mounted by pins that were pushed through the fins of the porpoises or the backs of the narwhals. Overall body condition seemed unaffected by the instrumentations. Macroscopical examination revealed that umbilicalization of the tissue surrounding the pins was almost complete. On one of the narwhals the reepithelialization created a closed tunnel where the pins were isolated from the subdermal tissue, however the reepithelialization was incomplete around the middle of the pin and a low‐grade inflammation increased with decreasing thickness of epidermis. The inflammation consisted of mononuclear cells, mainly lymphocytes. With increasing inflammation the number of neutrophils and macrophages increased. In the lymphoid follicular hyperplasia macrophages and a few neutrophils were found, in one case accompanied by Splendore‐Hoeppli material with radiating eosinophilic clubs and Gram‐positive cocci. Immunohistochemical staining of the cocci for Staphylococcus aureus was positive. The observations from the recaptured cetaceans suggest that the instrumentations caused only temporary and low‐grade inflammatory responses.     

Brain microabscesses in a porcine model of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> sepsis

Background

Sepsis caused by Staphylococcus aureus often leads to brain microabscesses in humans. Animal models of haematogenous brain abscesses would be useful to study this condition in detail. Recently, we developed a model of S. aureus sepsis in pigs and here we report that brain microabscesses develop in pigs with such induced S. aureus sepsis.Twelve pigs were divided into three groups. Nine pigs received an intravenous inoculation of S. aureus once at time 0 h (group 1) or twice at time 0 h and 12 h (groups 2 and 3). In each group the fourth pig served as control. The pigs were euthanized at time 12 h (Group 1), 24 h (Group 2) and 48 h (Group 3) after the first inoculation. The brains were collected and examined histopathologically.

Results

All inoculated pigs developed sepsis and seven out of nine pigs developed brain microabscesses. The microabscesses contained S. aureus and were located in the prosencephalon and mesencephalon. Chorioditis and meningitis occurred from 12 h after inoculation.

Conclusions

Pigs with experimental S. aureus sepsis often develop brain microabscesses. The porcine brain pathology mirrors the findings in human sepsis patients. We therefore suggest the pig as a useful animal model of the development of brain microabscesses caused by S. aureus sepsis.

     

A Gly98Val Mutation in the N-Myc Downstream Regulated Gene 1 (NDRG1) in Alaskan Malamutes with Polyneuropathy

The first cases of early-onset progressive polyneuropathy appeared in the Alaskan Malamute population in Norway in the late 1970s. Affected dogs were of both sexes and were ambulatory paraparetic, progressing to non-ambulatory tetraparesis. On neurologic examination, affected dogs displayed predominantly laryngeal paresis, decreased postural reactions, decreased spinal reflexes and muscle atrophy. The disease was considered eradicated through breeding programmes but recently new cases have occurred in the Nordic countries and the USA. The N-myc downstream-regulated gene (NDRG1) is implicated in neuropathies with comparable symptoms or clinical signs both in humans and in Greyhound dogs. This gene was therefore considered a candidate gene for the polyneuropathy in Alaskan Malamutes. The coding sequence of the NDRG1 gene derived from one healthy and one affected Alaskan Malamute revealed a non-synonymous G>T mutation in exon 4 in the affected dog that causes a Gly98Val amino acid substitution. This substitution was categorized to be “probably damaging” to the protein function by PolyPhen2 (score: 1.000). Subsequently, 102 Alaskan Malamutes from the Nordic countries and the USA known to be either affected (n = 22), obligate carriers (n = 7) or healthy (n = 73) were genotyped for the SNP using TaqMan. All affected dogs had the T/T genotype, the obligate carriers had the G/T genotype and the healthy dogs had the G/G genotype except for 13 who had the G/T genotype. A protein alignment showed that residue 98 is conserved in mammals and also that the entire NDRG1 protein is highly conserved (94.7%) in mammals. We conclude that the G>T substitution is most likely the mutation that causes polyneuropathy in Alaskan Malamutes. Our characterization of a novel candidate causative mutation for polyneuropathy offers a new canine model that can provide further insight into pathobiology and therapy of human polyneuropathy. Furthermore, selection against this mutation can now be used to eliminate the disease in Alaskan Malamutes.     

Local osteogenic expression of cyclooxygenase-2 and systemic response in porcine models of osteomyelitis

It is suggested that cyclooxygenase 2 (COX-2) derived prostaglandins contributes to the progressive bone loss seen in osteomyelitis lesions. In the present study we examined the expression of COX-2 in bones from 23 pigs with experimental osteomyelitis. Osteomyelitis was induced with Staphylococcus aureus and groups of animals were euthanized following 6 h, 12 h, 24 h, 2 days, 5 days, 11 days and 15 days, respectively. Expression of COX-2 was evaluated immunohistochemically and combined with characterization of morphological changes in bone tissue. Furthermore, the serum concentrations of alkaline phosphatase and haptoglobin were measured. Extensive COX-2 expression by osteoblasts was present 2 days after inoculation together with many activated osteoclasts. Simultaneously, the serum concentration of alkaline phosphatase decreased whereas the haptoglobin concentration increased. This is the first in vivo study showing an early wave of COX-2 mediated bone resorption during osteomyelitis. Therefore, treatment aiming to reduce the break down of bone tissue directed by the COX-2 pathway might be suggested early in the course of the disease.     

Two decades of biomonitoring polar bear health in Greenland: a review

Background

Different animal models are used as fracture models in orthopaedic research prior to implant use in humans, although biomechanical forces can differ to a great extend between species due to variable anatomic conditions, particularly with regard to the gait. The rabbit is an often used fracture model, but biomechanical data are very rare. The objective of the present study was to measure axial forces, bending moments, and bending axis directly in the rabbit tibia in vivo. The following hypothesis was tested: Axial forces and bending moments in the mid-diaphysis of rabbit tibia differ from other experimental animals or indirectly calculated data.

Methods

A minifixateur system with 4 force sensors was developed and attached to rabbit tibia (n = 4), which were subsequently ostectomised. Axial forces, bending moments and bending angles were calculated telemetrically during weight bearing in motion between 6 and 42 days post operation.

Results

Highest single values were 201% body weight [% bw] for axial forces and 409% bw cm for bending moments. Whereas there was a continous decrease in axial forces over time after day 10 (P = 0.03 on day 15), a decrease in bending moments was inconsistent (P = 0.03 on day 27). High values for bending moments were frequently, but not consistently, associated with high values for axial forces.

Conclusion

Axial forces in rabbit tibia exceeded axial forces in sheep, and differed from indirectly calculated data. The rabbit is an appropriate fracture model because axial loads and bending moments in rabbit tibia were more closely to human conditions than in sheep tibia as an animal model.     

A pig model of acute Staphylococcus aureus induced pyemia

Background

Sepsis caused by Staphylococcus aureus constitutes an important cause of morbidity and mortality in humans, and the incidence of this disease-entity is increasing. In this paper we describe the initial microbial dynamics and lesions in pigs experimentally infected with S. aureus, with the aim of mimicking human sepsis and pyemia.

Methods

The study was conducted in anaesthetized and intravenously inoculated pigs, and was based on bacteriological examination of blood and testing of blood for IL-6 and C-reactive protein. Following killing of the animals and necropsy bacteriological and histological examinations of different organs were performed 4, 5 or 6 h after inoculation.

Results

Clearance of bacteria from the blood was completed within the first 2 h in some of the pigs and the highest bacterial load was recorded in the lungs as compared to the spleen, liver and bones. This probably was a consequence of both the intravenous route of inoculation and the presence of pulmonary intravascular macrophages. Inoculation of bacteria induced formation of acute microabscesses in the lungs, spleen and liver, but not in the kidneys or bones. No generalized inflammatory response was recorded, i.e. IL-6 was not detected in the blood and C-reactive protein did not increase, probably because of the short time course of the study.

Conclusion

This study demonstrates the successful induction of acute pyemia (microabscesses), and forms a basis for future experiments that should include inoculation with strains of S. aureus isolated from man and an extension of the timeframe aiming at inducing sepsis, severe sepsis and septic shock.