Welfare of extensively managed Swedish Gotland ponies.

It has been suggested that grazing horses could be used as a credible tool for landscape conservation which would, at the same time, improve horse welfare as opposed to conventional housing. A study was conducted between May 2014 and April 2015 on 12 one year old Gotland ponies managed extensively without supplementary feed. Monthly animal welfare assessments (n = 13) revealed welfare issues in most of the horses, i.e. low body condition score (BCS < 3/5), recurring poor skin condition in 11/12 horses and ocular discharge in 7/12 horses. At the end of the study, compared to the beginning, chafing and poor skin condition increased while coat condition improved. A correlation was found between a negative reaction (score > 0) in the human approach test and BCS < 3 and ocular discharge. Avoidance Distance test values were correlated with faecal parasite counts (> 350 eggs per gram [EPG]). These results indicate that the horses had acceptable welfare during late spring/summer (May-September) and that some horses required additional feed during winter. The animal welfare protocol proved to be an efficient tool for monitoring welfare. The results showed that factors important for extensive management are: daily monitoring; enclosures that provide sufficient feed; access to recovery enclosure; and habituation of horses to human approach.

The sGC Activator Runcaciguat Has Kidney Protective Effects and Prevents a Decline of Kidney Function in ZSF1 Rats.

Chronic kidney disease (CKD) progression is associated with persisting oxidative stress, which impairs the NO-sGC-cGMP signaling cascade through the formation of oxidized and heme-free apo-sGC that cannot be activated by NO. Runcaciguat (BAY 1101042) is a novel, potent, and selective sGC activator that binds and activates oxidized and heme-free sGC and thereby restores NO-sGC-cGMP signaling under oxidative stress. Therefore, runcaciguat might represent a very effective treatment option for CKD/DKD. The potential kidney-protective effects of runcaciguat were investigated in ZSF1 rats as a model of CKD/DKD, characterized by hypertension, hyperglycemia, obesity, and insulin resistance. ZSF1 rats were treated daily orally for up to 12 weeks with runcaciguat (1, 3, 10 mg/kg/bid) or placebo. The study endpoints were proteinuria, kidney histopathology, plasma, urinary biomarkers of kidney damage, and gene expression profiling to gain information about relevant pathways affected by runcaciguat. Furthermore, oxidative stress was compared in the ZSF1 rat kidney with kidney samples from DKD patients. Within the duration of the 12-week treatment study, kidney function was significantly decreased in obese ZSF1 rats, indicated by a 20-fold increase in proteinuria, compared to lean ZSF1 rats. Runcaciguat dose-dependently and significantly attenuated the development of proteinuria in ZSF1 rats with reduced uPCR at the end of the study by -19%, -54%, and -70% at 1, 3, and 10 mg/kg/bid, respectively, compared to placebo treatment. Additionally, average blood glucose levels measured as HbA1C, triglycerides, and cholesterol were increased by five times, twenty times, and four times, respectively, in obese ZSF1 compared to lean rats. In obese ZSF1 rats, runcaciguat reduced HbA1c levels by -8%, -34%, and -76%, triglycerides by -42%, -55%, and -71%, and cholesterol by -16%, -17%, and -34%, at 1, 3, and 10 mg/kg/bid, respectively, compared to placebo. Concomitantly, runcaciguat also reduced kidney weights, morphological kidney damage, and urinary and plasma biomarkers of kidney damage. Beneficial effects were accompanied by changes in gene expression that indicate reduced fibrosis and inflammation and suggest improved endothelial stabilization. In summary, the sGC activator runcaciguat significantly prevented a decline in kidney function in a DKD rat model that mimics common comorbidities and conditions of oxidative stress of CKD patients. Thus, runcaciguat represents a promising treatment option for CKD patients, which is in line with recent phase 2 clinical study data, where runcaciguat showed promising efficacy in CKD patients (NCT04507061).

Effects of Finerenone Combined with Empagliflozin in a Model of Hypertension-Induced End-Organ Damage.

INTRODUCTION: The nonsteroidal mineralocorticoid receptor (MR) antagonist finerenone and sodium-glucose cotransporter-2 (SGLT2) inhibitors have demonstrated clinical benefits in CKD patients with type 2 diabetes. Clinical data analyzing the potential value of a combination therapy are currently limited. We therefore investigated cardiorenal protection of respective mono- and combination therapy in a preclinical model of hypertension-induced end-organ damage. METHODS: Cardiovascular (CV) morbidity and mortality were studied in hypertensive, N(ω)-nitro-L-arginine methyl ester-treated, renin-transgenic (mRen2)27 rats. Rats (10- to 11-week-old females, n = 13-17/group) were treated once daily orally for up to 7 weeks with placebo, finerenone (1 and 3 mg/kg), empagliflozin (3 and 10 mg/kg), or a combination of the respective low doses. Key outcome parameters included mortality, proteinuria, plasma creatinine and uric acid, blood pressure, and cardiac and renal histology. RESULTS: Placebo-treated rats demonstrated a 50% survival rate over the course of 7 weeks. Drug treatment resulted in variable degrees of survival benefit, most prominently in the low-dose combination group with a survival benefit of 93%. Monotherapies of finerenone or empagliflozin dose-dependently reduced proteinuria, while low-dose combination revealed an early, sustained, and over-additive reduction in proteinuria. Empagliflozin induced a strong and dose-dependent increase in urinary glucose excretion which was not influenced by finerenone coadministration in the combination arm. Low-dose combination but not respective low-dose monotherapies significantly reduced plasma creatinine and plasma uric acid after 6 weeks. Treatment with finerenone and the low-dose combination significantly decreased systolic blood pressure after 5 weeks. There was a dose-dependent protection from cardiac and kidney fibrosis and vasculopathy with both agents, while low-dose combination therapy was more efficient than the respective monotherapy dosages on most cardiorenal histology parameters. DISCUSSION/CONCLUSIONS: Nonsteroidal MR antagonism by finerenone and SGLT2 inhibition by empagliflozin confer CV protection in preclinical hypertension-induced cardiorenal disease. Combination of these 2 independent modes of action at low dosages revealed efficacious reduction in important functional parameters such as proteinuria and blood pressure, plasma markers including creatinine and uric acid, cardiac and renal lesions as determined by histopathology, and mortality indicating a strong potential for combined clinical use in cardiorenal patient populations.

Direct Blood Pressure-Independent Anti-Fibrotic Effects by the Selective Nonsteroidal Mineralocorticoid Receptor Antagonist Finerenone in Progressive Models of Kidney Fibrosis.

INTRODUCTION: The nonsteroidal mineralocorticoid receptor (MR) antagonist finerenone and sodium-glucose cotransporter-2 (SGLT2) inhibitors have demonstrated clinical benefits in chronic kidney disease patients with type 2 diabetes. Precise molecular mechanisms responsible for these benefits are incompletely understood. Here, we investigated potential direct anti-fibrotic effects and mechanisms of nonsteroidal MR antagonism by finerenone or SGLT2 inhibition by empagliflozin in 2 relevant mouse kidney fibrosis models: unilateral ureter obstruction and sub-chronic ischemia reperfusion injury. METHODS: Kidney fibrosis was induced in mice via unilateral ureteral obstruction or ischemia. In a series of experiments, mice were treated orally with the MR antagonist finerenone (3 or 10 mg/kg), the SGLT2 inhibitor empagliflozin (10 or 30 mg/kg), or in a direct comparison of both drugs. Interstitial myofibroblast accumulation was quantified via alpha-smooth muscle actin and interstitial collagen deposition via Sirius Red/Fast Green staining in both models. Secondary analyses included the assessment of inflammatory cells, kidney mRNA expression of fibrotic markers as well as functional parameters (serum creatinine and albuminuria) in the ischemic model. Blood pressure was measured via telemetry in healthy conscious compound-treated animals. RESULTS: Finerenone dose-dependently decreased pathological myofibroblast accumulation and collagen deposition with no effects on systemic blood pressure and inflammatory markers in the tested dose range. Reduced kidney fibrosis was paralleled by reduced kidney plasminogen activator inhibitor-1 (PAI-1) and naked cuticle 2 (NKD2) expression in finerenone-treated mice. In contrast, treatment with empagliflozin strongly increased urinary glucose excretion in both models and reduced ischemia-induced albuminuria but had no effects on kidney myofibroblasts or collagen deposition. DISCUSSION/CONCLUSION: Finerenone has direct anti-fibrotic properties resulting in reduced myofibroblast and collagen deposition accompanied by a reduction in renal PAI-1 and NKD2 expression in mouse models of progressive kidney fibrosis at blood pressure-independent dosages.

Assessing the Use of the sGC Stimulator BAY-747, as a Potential Treatment for Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder, affecting one in 3500 to 5000 boys worldwide. The NO-sGC-cGMP pathway plays an important role in skeletal muscle function, primarily by improving blood flow and oxygen supply to the muscles during exercise. In fact, PDE5 inhibitors have previously been investigated as a potential therapy for DMD, however, a large-scale Phase III clinical trial did not meet its primary endpoint. Since the efficacy of PDE5i is dependent on sufficient endogenous NO production, which might be impaired in DMD, we investigated if NO-independent sGC stimulators, could have therapeutic benefits in a mouse model of DMD. Male mdx/mTRG2 mice aged six weeks were given food supplemented with the sGC stimulator, BAY-747 (150 mg/kg of food) or food alone (untreated) ad libitum for 16 weeks. Untreated C57BL6/J mice were used as wild type (WT) controls. Assessments of the four-limb hang, grip strength, running wheel and serum creatine kinase (CK) levels showed that mdx/mTRG2 mice had significantly reduced skeletal muscle function and severe muscle damage compared to WT mice. Treatment with BAY-747 improved grip strength and running speed, and these mice also had reduced CK levels compared to untreated mdx/mTRG2 mice. We also observed increased inflammation and fibrosis in the skeletal muscle of mdx/mTRG2 mice compared to WT. While gene expression of pro-inflammatory cytokines and some pro-fibrotic markers in the skeletal muscle was reduced following BAY-747 treatment, there was no reduction in infiltration of myeloid immune cells nor collagen deposition. In conclusion, treatment with BAY-747 significantly improves several functional and pathological parameters of the skeletal muscle in mdx/mTRG2 mice. However, the effect size was moderate and therefore, more studies are needed to fully understand the potential treatment benefit of sGC stimulators in DMD.

Finerenone, a novel selective nonsteroidal mineralocorticoid receptor antagonist protects from rat cardiorenal injury.

Pharmacological blockade of the mineralocorticoid receptor (MR) ameliorates end-organ damage in chronic heart failure. However, the clinical use of available steroidal MR antagonists is restricted because of concomitant hyperkalemia especially in patients with diminished kidney function. We have recently identified a novel nonsteroidal MR antagonist, finerenone, which uniquely combines potency and selectivity toward MR. Here, we investigated the tissue distribution and chronic cardiorenal end-organ protection of finerenone in comparison to the steroidal MR antagonist, eplerenone, in 2 different preclinical rat disease models. Quantitative whole-body autoradiography revealed that [C]-labeled finerenone equally distributes into rat cardiac and renal tissues. Finerenone treatment prevented deoxycorticosterone acetate-/salt-challenged rats from functional as well as structural heart and kidney damage at dosages not reducing systemic blood pressure. Finerenone reduced cardiac hypertrophy, plasma prohormone of brain natriuretic peptide, and proteinuria more efficiently than eplerenone when comparing equinatriuretic doses. In rats that developed chronic heart failure after coronary artery ligation, finerenone (1 mg·kg·d), but not eplerenone (100 mg·kg·d) improved systolic and diastolic left ventricular function and reduced plasma prohormone of brain natriuretic peptide levels. We conclude that finerenone may offer end-organ protection with a reduced risk of electrolyte disturbances.

Finnish and Swedish riding school pupils' motivation towards participation in non-riding education.

Many horse enthusiasts have insufficient knowledge about horse behavior and welfare (BW) and learning and human-horse communication (LC), which poses a risk for both horse welfare and human safety. The main objective of this study was to investigate why riding school pupils participate or do not participate in non-riding education in BW and LC, using Self-determination theory (SDT). SDT posits that the quality of motivation is related to the individual's basic psychological needs. A convenience sample of 568 riding school pupils from Finland and Sweden completed an online questionnaire. The results showed that forty percent of the riding schools offered education in BW, and thirty-two in LC. Twenty-seven percent of the respondents participated in education in BW, and twenty-five in LC at their riding school. The respondents were autonomously motivated to participate in education, i.e., they would participate because it is interesting and personally important. Perceived needs satisfaction at the riding school predicted autonomous motivation to participate. Education was offered to a greater extent in Swedish riding schools and Swedish respondents participated more often, as well as experienced more autonomous motivation, relatedness and competence satisfaction compared with Finnish respondents. To our knowledge, this study is the first to explore riding school pupils' motivation towards non-riding education.

Mineralocorticoid receptor-mediated DNA damage in kidneys of DOCA-salt hypertensive rats.

Epidemiological studies exploring the connection between hypertension and cancer demonstrate a higher cancer incidence, especially of kidney cancer, and a higher cancer mortality in hypertensive patients. Hormones elevated in hypertension, i.e., aldosterone and angiotensin II, which exert genotoxic effects in vitro, could contribute to carcinogenesis in hypertension. The present study was conducted to investigate the possible DNA-damaging effect of aldosterone receptor activation in vivo. Crl:CD (Sprague-Dawley) rats were treated for 6 wk with desoxycorticosterone acetate (DOCA) and salt to induce a mineralocorticoid-dependent hypertension. DOCA-salt treatment caused increased blood pressure (+26 mmHg) compared to untreated rats, elevated markers of kidney failure (up to 62-fold for Kim-1), and the induction of several proinflammatory genes and proteins (up to 2.6-fold for tissue MCP-1). The mineralocorticoid receptor (MR) antagonist spironolactone (MR IC(50) 24 nM) and the novel nonsteroidal antagonist BR-4628 (MR IC(50) 28 nM) decreased these damage markers. DOCA-salt treatment also caused 8.8-fold increased structural DNA damage, determined with the comet assay, double-strand breaks (3.5-fold), detected immunohistochemically, and oxidative stress. Furthermore, the oxidatively modified mutagenic DNA base 7,8-dihydro-8-oxo-guanine (8-oxodG), quantified by LC-MS/MS, was almost 2-fold higher in DOCA-salt-treated kidneys. Our results suggest a mutagenic potential of high mineralocorticoid levels, frequent in hypertensive individuals.

Urinary miRNA Profiles in Chronic Kidney Injury-Benefits of Extracellular Vesicle Enrichment and miRNAs as Potential Biomarkers for Renal Fibrosis, Glomerular Injury, and Endothelial Dysfunction.

Micro-RNAs (miRNAs) are regulators of gene expression and play an important role in physiological homeostasis and disease. In biofluids, miRNAs can be found in protein complexes or in extracellular vesicles (EVs). Altered urinary miRNAs are reported as potential biomarkers for chronic kidney disease (CKD). In this context, we compared established urinary protein biomarkers for kidney injury with urinary miRNA profiles in obese ZSF1 and hypertensive renin transgenic rats. Additionally, the benefit of urinary EV enrichment was investigated in vivo and the potential association of urinary miRNAs with renal fibrosis in vitro. Kidney damage in both rat models was confirmed by histopathology, proteinuria, and increased levels of urinary protein biomarkers. In total, 290 miRNAs were elevated in obese ZSF1 rats compared with lean controls, whereas 38 miRNAs were altered in obese ZSF1 rats during 14-26 weeks of age. These 38 miRNAs correlated better with disease progression than established urinary protein biomarkers. MiRNAs increased in obese ZSF1 rats were associated with renal inflammation, fibrosis, and glomerular injury. Eight miRNAs were also changed in urinary EVs of renin transgenic rats, including one which might play a role in endothelial dysfunction. EV enrichment increased the number and detection level of several miRNAs implicated in renal fibrosis in vitro and in vivo. Our results show the benefit of EV enrichment for miRNA detection and the potential of total urine and urinary EV-associated miRNAs as biomarkers of altered kidney physiology, renal fibrosis and glomerular injury, and disease progression in hypertension and obesity-induced CKD.

Rein tension in harness trotters during on-track exercise.

Horseracing is under public scrutiny with increasing demands to safeguard horse welfare. It is accepted that, as a result of bit pressure and/or equipment, mouth lesions accompany many types of horse use, including racing. However, there are currently no data available on the range of bit pressures in driven trotters. Our aim was to investigate whether rein tension (RT, proxy for bit pressures) differs among gaits, between tempo within gait, between horses and drivers, and between left/right reins. Standardbreds (n = 9), driven by experienced drivers (n = 11), performed exercise tests on a racetrack (cross-over design; total 31 tests, data available from 26 tests). Horses' motion symmetry was measured before tests (trotting in hand). Rein tension, speed and heart rate were measured during exercise. A moving-window filter was applied to RT raw data. Median, maximum and interquartile range for the estimated stride median RT were determined for each rein (left/right) and segment: walk; circling in slow trot followed by transition to faster trot; fast (racing) trot; and slowing down to walk. Mixed models were used for statistical analysis. Least square means for segment median RT ranged between 17-19 N in walk, 34-40 N during circling-accelerating, 51-62 N in fast trot, and 53-71 N for slowing down. Segment maximum RT was between 60-81 N in walk, 104-106 N during circling-accelerating, 72-86 N in fast trot, and 86-129 N during slowing down. Interquartile ranges were between 7-9 N in walk, 28-31 N during circling-accelerating, 8-10 N in fast trot, and 12-18 N for slowing down. Hind limb asymmetry exceeded the recommended threshold in three horses and was associated with higher median (48 N) and maximum (106 N) RT than symmetric horses (29 N and 73 N, respectively, p < 0.01). Consistent left-right asymmetry in RT was more common among horses than among drivers. Rein tension increased with increasing heart rate (p ≤ 0.0006). Rein tensions were higher than those reported during riding or in horses worked from the ground. The findings of high RT, taken together with the high reported prevalence of oral injuries in harness trotters, call for further research into RT, motion symmetry and use of equipment.

From the Horse's Perspective: Investigating Attachment Behaviour and the Effect of Training Method on Fear Reactions and Ease of Handling-A Pilot Study.

The study investigated equine responses to novelty and handling, aiming to reveal whether horse-human relationships reflect criteria of an attachment bond. Twelve adult Standardbreds were subjected to a fear-eliciting test (novel objects presented close to two humans) and a handling test (being led passing novel objects) to study attachment-related behaviours and ease of handling. The tests were performed both before (pre-test) and after (post-test) horses had been trained by the same female handler (10 sessions of 15 min). Horses were assigned to three groups of four, each of which underwent different operant conditioning protocols: negative reinforcement (NR; pressure, release of lead, and whip tap signals) or combined NR with either positive reinforcement using food (PRf) or wither scratching (PRs). Results showed that neither familiarity of the person nor training method had a significant impact on the horses' behavioural responses in the post-tests. However, horses showed decreased heart rates between pre- and post-tests, which may indicate habituation, an effect of training per se, or that the presence of the familiar trainer served to calm the horses during the challenging situations. There were large individual variations among the horses' responses and further studies are needed to increase our understanding of horse-human relationships.

Runcaciguat, a novel soluble guanylate cyclase activator, shows renoprotection in hypertensive, diabetic, and metabolic preclinical models of chronic kidney disease.

Chronic kidney diseaQueryse (CKD) is associated with oxidative stress which can interrupt the nitric oxide (NO)/soluble guanylyl cyclase (sGC) signaling and decrease cyclic guanosine monophosphate (cGMP) production. Low cGMP concentrations can cause kidney damage and progression of CKD. The novel sGC activator runcaciguat targets the oxidized and heme-free form of sGC, restoring cGMP production under oxidative stress. The purpose of this study is to investigate if runcaciguat could provide an effective treatment for CKD. Runcaciguat was used for the treatment not only in rat CKD models with different etiologies and comorbidities, namely of hypertensive rats, the renin transgenic (RenTG) rat, and angiotensin-supplemented (ANG-SD) rat, but also in rats with diabetic and metabolic CKD, the Zucker diabetic fatty (ZDF) rat. The treatment duration was 2 to 42 weeks and runcaciguat was applied orally in doses from 1 to 10 mg/kg/bid. In these different rat CKD models, runcaciguat significantly reduced proteinuria (urinary protein to creatinine ratio; uPCR). These effects were also significant at doses which did not or only moderately decrease systemic blood pressure. Moreover, runcaciguat significantly decreased kidney injury biomarkers and attenuated morphological kidney damages. In RenTG rats, runcaciguat improved survival rates and markers of heart injury. These data demonstrate that the sGC activator runcaciguat exhibits cardio-renal protection at doses which did not reduce blood pressure and was effective in hypertensive as well as diabetic and metabolic CKD models. These data, therefore, suggest that runcaciguat, with its specific mode of action, represents an efficient treatment approach for CKD and associated CV diseases.

Effect of work on body language of ranch horses in Brazil.

The horses' responses to exercise are commonly monitored using physiological variables, nonetheless physical and mental states can also be expressed through body language. The aims of this study were: (i) to identify how facial expressions and other behavioural variables change in ranch horses after a routine workday, and (ii) to investigate if these changes can be used as indicators of physical tiredness by relating them to known variables of physical fitness and workload. Fourteen crossbred ranch horses were assessed pre- and post-workday on two farms, recording the body language, physiological and workload variables. Statistical analysis consisted of four stages: (i) comparisons between the sampling times (pre- vs post-workday) using linear mixed-effects models with repeated measures and a paired Wilcoxon test; (ii) selection of the most powerful variables by applying Kaiser-Meyer-Olkin test and principal components analyses (PCA); (iii) evaluations of the relationships within these selected variables utilizing PCA and Spearman rank coefficients; and (iv) identifying a critical level of the most robust behavioural indicators using a non-hierarchical cluster analysis. Results showed that after a workday the horses increased the frequency/duration of body language indicative of resting. They also decreased the frequency/duration of body language indicative of attention and movements to avoid flies. However, some of these behaviours are also shown when horses are in pain, leading us to suggest that some ranch horses were probably experiencing a combination of of tiredness and slight soreness. Of particular interest, because of the ease with which it can be assessed on the farm and generalized to other situations, we suggest that the frequency of shifting weight between the forelegs has potential to be used as an indicator of physical tiredness in horses. The results can also be used in the development of tools to improve the welfare of ranch horses as well as horses used in other activities, although more research is needed to validate this assumption.

A Novel Model of Chronic Kidney Disease in Rats: Dietary Adenine in Combination with Unilateral Nephrectomy.

BACKGROUND: Adenine at 0.75% in the diet (AD) triggers renal impairment in rats. This model of kidney disease is largely reversible when AD feeding is stopped. Testing of novel drugs parallel to AD administration may result in unwanted interference. OBJECTIVES: We hypothesized that combining AD with unilateral nephrectomy (UNx) would result in progressive chronic kidney disease (CKD) even after cessation of AD. METHODS: In an explorative study, 16 rats with UNx (AD-1K rats) and 10 sham-operated rats (AD-2K rats) received AD-supplemented feed for 3 weeks, followed by 4 weeks of standard chow. Ten sham-operated rats receiving only standard chow served as controls. Laboratory parameters in blood and urine were frequently assessed during and after cessation of AD feeding. Comprehensive pathological examinations were performed in all rats at the end of the experiment. RESULTS: Rats with UNx were more affected by impaired glomerular filtration rate, anemia, hyperphosphatemia, and hypocalcemia. After cessation of AD feeding, recovery was poorest in AD-1K rats, paralleled by increased proteinuria indicative of progressive CKD. Scores in histopathological damage of the kidneys indicative of CKD were seen in both AD-fed groups, with key parameters being more affected in AD-1K rats. Histopathological changes in the heart were most prominent in AD-1K rats. CONCLUSIONS: Combining AD feeding with UNx provides a time window after cessation of AD feeding for the testing of drugs without interference. Our findings in rats may have implications for research in other target animal species as well.

Reactive and proliferative changes of splenic reticulum cells of rats investigated with special staining methods and immunohistochemistry.

In the rat spleen, reactive and proliferative changes of the reticulum cells are rare events and seem to occur almost exclusively in the red pulp. The normal structure of the splenic reticulum cell and fiber lattice and examples of spontaneous and induced pathological alterations were investigated by immunohistochemistry (smooth muscle-actin, vimentin, S100 and proliferating cell nuclear antigen) and special stains for extracellular fibers (silver impregnation, azan). In response to congestion, systemic tumor growth or treatment with a hematotoxic compound, the scaffold cells increased either their contractile properties or their production of extracellular fibers. Primary focal hyperplasias of stromal cells which had developed without obvious cause were characterized by vanishing of sinuses, increased fiber content, increased expression of sm-actin or foci of lipomatosis. The borders of focal hyperplasias were indistinct and they did not infiltrate the white pulp compartments. Neoplasms of the stromal reticulum cells resembled soft tissue tumors in other organs. Specific tumor entities as described in other species have so far not been observed in the rat.

ESTP comments on the draft updated OECD test guideline 407.

The revision of the OECD TG 407 test guideline (repeated dose 28-day oral toxicity study in rodents) focuses on endpoints to detect endocrine activities of chemicals. The new endpoints are likely to influence other previously established core endpoints of this study type. An expert group of pathologists and toxicologists within the European Society of Toxicologic Pathologists (ESTP) has contributed to the scientific discussion of the draft guideline. The advantages and disadvantages of methodical changes as necropsy of all females in dioestrus, blood collection for clinical chemistry and haematology at the same cycle stage, weighing of the thyroid gland and separate weighing of ventral and dorsolateral lobes of the prostate are considered. Possible alternatives are pointed out covering scientific as well as practical aspects.

Evaluation of the rodent Hershberger bioassay using three reference (anti)androgens.

Under the umbrella of the Organization for Economic Cooperation and Development (OECD) the rodent Hershberger bioassay is being validated as an in vivo screen for the detection of (anti)androgens. As part of the validation work we studied trenbolone (TREN), 1,1-bis-(4-chlorophenyl)-2,2-dichloroethylene (p,p'-DDE) and vinclozolin (VIN). Oral intubation of castrated rats with TREN [0.3-1.5-8-40 mg kg(-1) body weight (b.w.)] for ten days increased androgen-sensitive tissue weights (ASTW) at the high dose. p,p'-DDE (5-16-50-160 mg kg(-1) b.w.) and VIN (0-3-10-30-100 mg kg(-1) b.w.) orally administered for ten days produced a dose-dependent decrease in ASTW in castrated rats subcutaneously supplemented with testosterone propionate (0.4 mg kg(-1) b.w.). p,p'-DDE also strongly increased liver weights and induced hepatocellular hypertrophy and thyroid follicular cell hypertrophy that was most likely mediated by liver enzyme induction. Our data strongly suggest that the OECD protocol of the rodent Hershberger bioassay describes a sensitive in vivo screen, capable of detecting weakly active (anti)androgens. Furthermore, our data may also indicate that thyroid effects could be assessed, if the protocol is amended accordingly.

24-h sheltering behaviour of individually kept horses during Swedish summer weather.

BACKGROUND: Provision of shelter for horses kept on summer pasture is rarely considered in welfare guidelines, perhaps because the benefits of shelter in warm conditions are poorly documented scientifically. For cattle, shade is a valued resource during summer and can mitigate the adverse effects of warm weather on well-being and performance. We found in a previous study that horses utilized shelters frequently in summer. A shelter with a roof and closed on three sides (shelter A) was preferred and can reduce insect pressure whereas a shelter with roof and open on three sides was not utilized. However, shelter A restricts the all-round view of a horse, which may be important for horses as flight animals. Therefore, we studied whether a shelter with roof, where only the upper half of the rear wall was closed (shelter B), would be utilized while maintaining insect protection properties and satisfying the horses' sense for security. A third shelter was offered with walls but no roof (shelter C) to evaluate whether the roof itself is an important feature from the horse's perspective. Eight Warmblood horses were tested each for 2 days, kept individually for 24 h in two paddocks with access to shelters A and B, or shelters A and C, respectively. Shelter use was recorded continuously during the night (1800-2400 h, 0200-0600 h) and the following day (0900-1600 h), and insect defensive behaviour (e.g., tail swish) in instantaneous scan samples at 5-min intervals during daytime. RESULTS: Seven horses used both shelters A and B, but when given the choice between shelters A and C, shelter C was scarcely visited. There was no difference in duration of shelter use between night (105.8 ± 53.6 min) and day (100.8 ± 53.8, P = 0.829). Daytime shelter use had a significant effect on insect defensive behaviours (P = 0.027). The probability of performing these behaviours was lowest when horses used shelter A compared to being outside (P = 0.038). CONCLUSIONS: Horses only utilized shelters with a roof whilst a shelter with roof and closed on three sides had the best potential to lower insect disturbance during daytime in summer.

Bilateral noncystic renal dysplasia in a Wistar-rat.

In a young Wistar rat a bilateral renal malformation was observed microscopically. Clinical chemistry gave no evidence of impaired kidney function. The kidney weight was slightly elevated and the kidneys showed no gross pathological changes. The lesion was located in the inner cortex of both kidneys and consisted of multiple foci of abnormal renal parenchyma similar to fetal kidney. Three components could be distinguished in the foci: primitive glomerular/tubular structures, tubules resembling collecting ducts and mesenchyme. For further characterisation, histological stains (H&E, PAS, Novotny) and immunohistochemistry (vimentin, pan-cytokeratin, S 100, proliferating cell nuclear antigen, and terminal desoxyribosyl-transferase mediated dUTP nick end labelling) were applied. The glomerular and tubular structures were hyperplastic and positive for proliferating cell nuclear antigen and vimentin. The collecting duct-like tubules were positive for pan-cytokeratin and gave no evidence of proliferation. The two epithelial components of the foci were surrounded by mesenchymal cells which extended also between the normal cortical tubules so that no clear demarcation was discernible. The mesenchymal cells were uniformly spindle-shaped and associated with reticulin fibers. Immunohistochemically they were vimentin-positive and non-proliferative. With terminal desoxyribosyl-transferase mediated dUTP nick end labelling (TUNEL) and S 100 all components were nearly negative. Based on morphology and immunohistochemistry this malformation containing structures derived from the ureteric bud and from the metanephric blastema associated with oligonephronia probably represents a noncystic renal dysplasia. Transition to neoplasia was not observed. A specific cause of this unusual developmental anomaly which was not previously reported in rats could not be determined.

Effects of infused administration volume on clinicochemical parameters and histopathology in vehicle control rats.

Infusion studies run during the last 10 years in three rat strains with different flow rates of saline as vehicle were compared as to a potential impact of infused volume on clinicochemical values and histopathology. There were no effects on white and red blood counts, except for a possible slight decrease of red blood cells with concomitant increase of reticulocytes after infusion of higher volumes over several weeks. There was also no volume-related effect on enzyme activities, metabolites/substrates, thyroid hormones and electrolytes in peripheral blood. As anticipated urinalyses proved an increase of excreted urine volume with increased infusion volume, while excreted protein, urine density and pH were not changed. Histopathological changes specific for intravenous infusion via indwelling catheter were seen in lungs, spleen and along the course of the catheter. Neither frequency nor severity of these findings was changed in relation to volume in the three rat strains examined. However, animal numbers are small for each rat strain examined and studies were run over almost 10 years. The impact of potential genetic drift, methodical changes and/or a change of infusion accessories (especially catheter) is unknown. An exchange of data and experience between laboratories should help differentiate between incidental and true effects.