A potentized homeopathic drug, Arsenicum Album 200, can ameliorate genotoxicity induced by repeated injections of arsenic trioxide in mice

Groundwater arsenic contamination has become a menacing global problem. No drug is available until now to combat chronic arsenic poisoning. To examine if a potentized homeopathic remedy, Arsenicum Album-200, can effectively combat chronic arsenic toxicity induced by repeated injections of Arsenic trioxide in mice, the following experimental design was adopted. Mice (Mus musculus) were injected subcutaneously with 0.016% arsenic trioxide at the rate of 1 ml/100 g body weight, at an interval of 7 days until they were killed at day 30, 60, 90 or 120 and were divided into three groups: (i) one receiving a daily dose of Arsenicum Album-200 through oral administration, (ii) one receiving the same dose of diluted succussed alcohol (Alcohol-200) and (iii) another receiving neither drug, nor succussed alcohol. The remedy or the placebo, as the case may be, was fed from the next day onwards after injection until the day before the next injection, and the cycle was repeated until the mice were killed. Two other control groups were also maintained: one receiving only normal diet, and the other receiving normal diet and succussed alcohol. Several toxicity assays, such as cytogenetical (chromosome aberrations, micronuclei, mitotic index, sperm head anomaly) and biochemical (acid and alkaline phosphatases, lipid peroxidation), were periodically made. Compared with controls, the drug fed mice showed reduced toxicity at statistically significant levels in respect of all the parameters studied, thereby indicating protective potentials of the homeopathic drug against chronic arsenic poisoning.     

Thirteen cases of tetanus in dogs

The records of 13 dogs with tetanus were reviewed, 12 of the dogs survived and were discharged, but the other died as a result of the acute onset of hyperthermia. Long-term follow up was available for 10 of the survivors of which nine were reported to be normal. One case was euthanased after a cervical spine fracture four months after it was discharged. Complications encountered during management included aspiration pneumonia in three cases, urinary tract infection in two cases, and upper respiratory tract obstruction, hiatal hernia, coxofemoral luxation, seizures and respiratory arrest in one case each. None of the dogs required ventilatory support, and the complications were managed successfully. The dogs were nursed intensively and monitored carefully.     

ULTRASONOGRAPHIC, GROSS, AND HISTOLOGIC EVALUATION OF A TENDINITIS DISEASE MODEL IN THE HORSE

Collagenase-induced lesions of the superficial digital flexor tendon were monitored using diagnostic ultrasound and subsequently compared with gross and microscopic findings. The postinjury sonograms demonstrated hypoechoic or anechoic areas corresponding to areas of hemorrhage and necrosis. The size of the lesion increased and the echogenicity decreased with increasing units of collagenase injected into the tendon. As postinjury time increased, echogenicity increased, which corresponded to fibrous-tissue invasion and collagen deposition. The extent of tendon injury produced correlated with the amount of collagenase injected into the tendon. Diagnostic ultrasound is useful in evaluating injury and healing in collagenase— induced tendinitis.     

Competition Horses Housed in Single Stalls (I): Behavior and Activity Patterns during Free Exercise According to Its Configuration

Although welfare of animals has become more important, housing horses in systems that limit natural behavior patterns is still widespread. Single stalls are the predominant housing system in Germany, especially for competition horses. Free exercise on fields or paddocks helps to improve welfare for the animals, but allowing it is not taken for granted by many horse keepers. In the present study, three common management practices were investigated with regard to their effect on the behavior of six competition horses housed in single stalls: 2-hour turnout after training in groups of two horses (group turnout [GT]), solitary turnout (ST) after training for 2 hours, and no turnout (NT) in addition to training. The aim of the study was to determine the differences in the horses’ behavior during turnout, training, and in the stable caused by the treatments. This article focuses on the behavior and activity patterns during turnout. The horses’ behavior was observed directly by the scan sampling method (5-minute interval), and the distance covered during turnout was measured by global positioning system devices. During turnout, the horses spent most of the time eating. After 1 hour of being turned out, eating decreased, and occupation, locomotion, and social interaction increased. When turnout was allowed in groups (GT), the horses spent more time in social interactions and less time walking and trotting than in ST. Therefore, they also covered a greater distance in ST than in GT. The distance covered in ST also showed a greater deviation both within and between days compared with GT.     

Exposure to Sarcocystis spp. in horses from Spain determined by Western blot analysis using Sarcocystis neurona merozoites as heterologous antigen

Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs.     

Gene expression in rabbit appendices infected with Eimeria coecicola

Eimeria coecicola causes intestinal coccidiosis in rabbits and, thereby, enormous economic losses in rabbit farms. Here, we investigate the final target site of E. coecicola, the appendix of rabbits, at the level of gene expression. Rabbits, orally infected with E. coecicola, begin to shed parasitic oocysts with their feces on day 5 p.i., and approximately 1.1 million oocysts are maximally shedded on day 7 p.i. At maximal shedding, the appendix has increased in size by about 2-3-folds and reveals increased hemorrhage which is associated with increases in nitrite/nitrate, malondialdehyde and catalase activity and a decrease in glutathione. Agilent 2-color oligo whole rabbit genome microarray, in combination with quantitative real-time PCR, detects 45 and 36 genes whose expression is more than 2-fold up- and down-regulated, respectively, by E. coecicola infection on day 7 p.i. The most dramatic increase by approximately 50-fold reveals the mRNA of the pro- and anti-inflammatory pleiotropic cytokine interleukin 6 (IL-6), whereas the largest decrease by approximately 13-fold is detected for mRNAs encoding for DBP, SULT3A1, CRP and glutathione-S transferase. Also, there are up- and down-regulations in the expression of genes encoding diverse regions of antibodies. Our data suggest that IL-6 plays a central role in the infection of the appendix of rabbits by E. coecicola, presumably involved in both pathological injuries, host defences and healing processes.     

Expression of receptors for luteinizing hormone,gastric-inhibitory polypeptide,and vasopressin in normal adrenal glands and cortisol-secreting adrenocortical tumors in dogs

Hypercortisolism caused by an adrenocortical tumor (AT) results from adrenocorticotropic hormone (ACTH)-independent hypersecretion of glucocorticoids. Studies in humans demonstrate that steroidogenesis in ATs may be stimulated by ectopic or overexpressed eutopic G protein-coupled receptors. We report on a screening of 23 surgically removed, cortisol-secreting ATs for the expression of receptors for luteinizing hormone (LH), gastric-inhibitory polypeptide (GIP), and vasopressin (V_1a, V_1b, and V₂). Normal adrenal glands served as control tissues. Abundance of mRNA for these receptors was quantified using quantitative polymerase chain reaction (QPCR), and the presence and localization of these receptors were determined by immunohistochemistry. In both normal adrenal glands and ATs, mRNA encoding for all receptors was present, although the expression abundance of the V_1b receptor was very low. The mRNA expression abundance for GIP and V₂ receptors in ATs were significantly lower (0.03 and 0.01, respectively) than in normal adrenal glands. The zona fasciculata of normal adrenal glands stained immunonegative for the GIP receptor. In contrast, islands of GIP receptor-immunopositive cells were detected in about half of the ATs. The zona fasciculata of both normal adrenal glands and AT tissue were immunopositive for LH receptor; in ATs in a homogenous or heterogenous pattern. In normal adrenal glands, no immunolabeling for V_1bR and V₂ receptor was present, but in ATs, V₂ receptor-immunopositive cells were detected. In conclusion, QPCR analysis did not reveal overexpression of LH, GIP, V_1a, V_1b, or V₂ receptors in the ATs. However, the ectopic expression of GIP and V₂ receptor proteins in tumorous zona fasciculata tissue may play a role in the pathogenesis of canine cortisol-secreting ATs.     

Tumor necrosis factor-α and acute-phase proteins in early pregnant ewes after challenge with peptidoglycan-polysaccharide

Bacterial infection shortly after mating interferes with establishment of pregnancy. Injection of peptidoglycan-polysaccharide (PG-PS), a component of gram-positive bacteria, into sheep on day 5 after mating reduces pregnancy rate. Experiments were designed to evaluate the acute-phase response (APR) in ewes to injection of PG-PS on day 5 after mating (day 0). Catheters were inserted into the jugular and posterior vena cava on day 4. On day 5, ewes were challenged with saline or 30 μg/kg body weight (BW) PG-PS (Exp 1) or 60 μg/kg BW PG-PS (Exp 2). Blood samples were collected every 15 min for 6 h (Exp 1) and every 15 min for 2 h, hourly for 12 h, and at 24, 36, and 48 h (Exp 2). Body temperature and clinical signs of infection were monitored in Exp 2. Plasma was assayed for concentrations of a pro-inflammatory cytokine, tumor necrosis factor-α (TNF-α); 2 APR proteins, serum amyloid A (SAA) and haptoglobin (Hp); and progesterone (P₄). Ewes injected with 60 μg/kg BW PG-PS exhibited fever, vaginal discharge, loss of appetite, and lethargy. After challenge with either 30 μg/kg or 60 μg/kg BW PG-PS, TNF-α increased in the posterior vena cava. Concentrations of SAA and Hp in the jugular increased after challenge with 60 μg/kg BW PG-PS. Only half (5/10) of the ewes treated with 60 μg/kg BW PG-PS had ultrasonically visible embryos, and none of them had functional corpora lutea (CL) (<1 ng/mL of P₄) on day 21. On the other hand, 8/9 (88.9%) control ewes had visible embryos and all had functional CL on day 21. Using logistic regression, pregnancy on day 21 was predicted to depend on concentrations of TNF-α and Hp on day 5 and concentration of P₄ on day 14. In summary, injection of PG-PS on day 5 after mating resulted in fever; increased concentrations of TNF-α, Hp, and SAA on the day of and the day after the PG-PS challenge; and decreased concentrations of P₄ on days 14 and 21. These factors were related to failure to establish pregnancy.     

Exfoliative cutaneous lupus erythematosus in German shorthaired pointer dogs: disease development,progression and evaluation of three immunomodulatory drugs (ciclosporin,hydroxychloroquine, and adalimumab) in a controlled environment

Six German shorthaired pointer dogs (two females, four males) with exfoliative cutaneous lupus erythematosus (ECLE) were studied in a controlled setting until disease progression necessitated euthanasia. During investigations into the heredity of disease, five dogs received immunomodulatory drugs to alleviate clinical signs (lameness, erythema, scaling, erosions/ulcers). One dog served as a control and received only baths and oral fatty acids. Four dogs received ciclosporin (5–10 mg/kg once daily) for 4.5 months to 2 years. Ciclosporin decreased erythema and arthralgia, but did not halt worsening of lesions. Three dogs received hydroxychloroquine (5–10 mg/kg once daily) for 8 weeks, 7 months, and 9 months, respectively, with no side effects. Hydroxychloroquine appeared to slow clinical progression in two dogs on extended treatment and normalized globulin levels in all three dogs while receiving the drug. Four dogs, including the control dog, were euthanized between 1 and 4.5 years of age. Two remaining male dogs received a tumour necrosis factor (TNF)‐α antagonist, adalimumab, at 0.5 mg/kg every 2 weeks for 8 weeks then weekly for 8 weeks. Serum TNF‐α levels were not significantly altered nor were quantifiable changes seen in skin lesions or lameness. Subsequently, the dogs were maintained on hydroxychloroquine for another year. This is the first study to evaluate the use of a TNF‐α inhibitor for canine lupus and the first to address the safety of long‐term administration of hydroxychloroquine, albeit in a small number of dogs. The study documents the progression of ECLE and generally poor response to therapy.