[Occurrance of antibodies against Leptospira in horses in Middle Germany]. / Vorkommen von Antikörpern gegen Leptospiren bei Pferden im mitteldeutschen Raum.

Aim of the study was to detect antibodies and potential risk factors for an infec- tion with Leptospira in horses in Middle Germany. Serum samples of 314 horses were examined retrospectively by microscopic agglutination test for the presence of antibodies against eight Leptospira serovars. In total, 17.2% (n = 54) of the horses were positive for one or more of the serovars analyzed. The most prevalent serovar was lcterohaemorrhagiae (11.1%), followed by serovar Bratislava (9.6 %) and Grippotyphosa (1.9%). Mares showed a significantly higher occurrence of antibodies (p < 0.05) than geldings or stallions. Horses used for breeding have a significantly lower risk than horses used in sport or horses used for leisure activity. There was also a significantly higher prevalence (p < 0.05) in summer than in the other seasons. No significant influence of breed, husbandry conditions and age on the antibody occurrence was observed (p > 0.05). The clinical chemical parameters did not differ significantly between horses with positive or negative Leptospira antibody result (p > 0.05). It became apparent that horses can be infected with Leptospira without developing of clinical symptoms.

[Detection of leptospira by culture of vitreous humor and detection of antibodies against leptospira in vitreous humor and serum of 225 horses with equine recurrent uveitis]. / Kultureller Nachweis von Leptospiren in Glaskörperflüssigkeit und Antikörpernachweis gegen Leptospiren in Glaskörperflüssigkeit und Serum von 225 Pferden mit equiner rezidivierender Uveitis (ERU).

In the ongoing discussion regarding the aetiopathogenesis of equine recurrent uveitis (ERU) it was the aim of the present study to elucidate the relationship of leptospira infection and ERU. In a population of 225 horses leptospira were examined in vitreous humor by culture and leptospira antibody were detected in vitreous humor and serum samples. Preoperative serum samples were collected from 221/225 ERU patients of different age, gender and breed. Undiluted vitreous humor was aseptically taken from 198/225 patients that underwent pars plana vitrectomy at the beginning of surgery and from 27/225 patients' eyeball after enucleation: Serum and vitreous humor were tested for specific leptospiral antibodies by microscopic agglutination test (MAT). Furthermore, vitreous humor was examined by culture. 20 patients which were euthanized due to a live-threatening disease other than ERU served as a control group. A total of 127/221 (57.5%) horses had serum antibodies (≥ 1:100). Most frequently antibodies against L. interrogans serovar Grippotyphosa were detected (79/127), followed by L. interrogans serovar lcterohaemorrhagiae (34/127) and L. interrogans serovar Bratislava (29/127). Only 79/225 horses (35.1%) had leptospiral antibodies in vitreous humor, in which L. interrogans serovar Grippotyphosa (67/79) was identified most frequently followed by L. interrogans serovar Pomona (18/79) and L. interrogans serovar lcterohaemorrhagiae (8/79) which was identified as single or multiple reaction. Isolation of leptospira from vitreous humor was positive in 34/212 horses (16%). 10/20 control horses had a positive antibody titer against leptospira in serum and 2/20 horses in vitreous humor, whereas there was no leptospira detected in culture. The result of 84% negative cultures from vitreous humor of 212 ERU patients is decisive for the diagnosis and therapy of ERU.

[Pituitary syndrome caused by neoplasia in a 17-month-old Holstein Friesian heifer]. / Hirnbasissyndrom infolge eines Tumors bei einer 17 Monate alten Deutsch-Holstein-Färse.

A 17-month-old Holstein Friesian heifer was presented after one day with dysphagia, slight paralysis of the tongue and swelling of the eyelids. Clinical examination of the animal revealed an extended posture of the head and neck, severely increased salivation, reduced lingual tone and mandibular paralysis with complete absence of the swallowing reflex. The right eye showed a drooping eyelid, mucopurulent discharge, exposure keratitis, corneal opacity and miosis. On the left side, a moderate exophthalmos and slight mucous discharge from the nostril were observed. Neurological examination revealed the absence of multiple cranial nerve reflexes suggesting a pituitary syndrome. On X-rays, a soft-tissue opacity with sharp margins and a diameter of approximately 5 cm was seen. It was located ventral to the ethmoid bone with possible intraneurocranial origin. Rhinoscopically, a mass located distal to the ethmoid bone with an uneven, slightly reddish surface partly covered by purulent exudate was visualised. Post-mortem examination of the euthanized animal confirmed neoplasia, which ranged from the fossa hypophysialis of the corpus ossis basisphenoidalis to the ethmoid bone. Histopathologic findings matched a small, round, blue cell tumour.

Impact of diet on 24-hour intragastric pH profile in healthy horses.

An electrode incorporated into a polyethylene hose was introduced under endoscopic control into the stomach of six fasting adult horses for long-lasting pH measurements. The intragastric pH was recorded every four seconds for a period of 24 hours. The Warmblood horses were assigned randomly to receive hay ad libitum (H group); 1.5 kg hay/100 kg BW/day and 1 kg concentrate/100 kg BW/ day (C group) or protocol C plus 75 g pectin-lecithin supplement/100 kg BW/day (P group). The horses were adapted to each diet for 14 days. The 24-hour median pH value for protocol H (2.69) was significantly lower compared to protocol C (3.35) and P (3.44) (p < 0.05). The horses in protocol P had a significant higher percentage (40.1 %) of 24-hour intragastric pH values ≥ 4 than in protocol C (36.2 %) or in protocol H (25.3 %) (p < 0.05).

[Evaluation of an immunochromatographic dipstick test for the assessment of tetanus immunity in horses]. / Evaluierung eines Schnelltestes zur Feststellung des Tetanus-Immunstatus bei Pferden.

Knowledge of tetanus immunity in equine patients is crucial in cases of injuries, elective surgeries, or when effective vaccination protocols are to be designed. The Fassisi® TetaCheck is a stall-side rapid test which was developed to address these issues. The purpose of the present study was to evaluate its performance parameters. To this end, the qualitative test results obtained by two blinded observers were compared to tetanus toxoid antibody levels from 99 serum samples, measured with a double antigen ELISA. Additionally the colour intensities of the test window were quantified using a camera and photo editing software. Assuming that the protective level of tetanus toxoid antibodies is ≥ 0.1 IE/ml, the tetanus quick stick (TQS) showed a sensitivity of 83.6% and a specificity of 100%. almost perfect (K = 0.88). Exchanging the observer did not affect the interpretation of theTQS (K = 0.80; K = 0.84). The definition of five distinct colour intensities of the "test window" enabled a clear differentiation of unprotected individuals from those with a protective immunity. There was a linear relationship between the objectively measured colour intensities and the tetanus toxoid antibody concentration (r2 = 0.74). The TQS thus proved to be a robust and reliable test in the stall-side assessment of tetanus immunity in horses. Its implementation in equine daily practice can help to avoid unnecessary immunizations in adult horses and therefore minimize vaccination side effects.

Influence of laxatives on gastric emptying in healthy warmblood horses evaluated with the acetaminophen absorption test.

The use of laxatives is crucial in the treatment of horses with caecal or large colon impaction.To reach the large intestinal contents and resolve the impaction, laxatives must leave the stomach and pass through the small intestine.The aim of this study was to prove whether isotonic solutions of saline cathartics do not affect gastric emptying rate in contrast to hypertonic solutions. Six, fasted, healthy, adult Warmblood horses were used in a randomized study design with 1.8% sodium sulfate (1.8% Na2SO4), 4.2% magnesium sulfate (4.2% MgSO4), 25% sodium sulfate (25% Na2SO4), 25% magnesium sulfate (25% MgSO4) and water at either 20 ml/kg BW (Water 20) or 4 ml/kg BW (Water 4), administered via nasogastric intubation. For indirect measurement of liquid-phase gastric emptying, the liquid-phase passage marker acetaminophen (20 mg/kg BW in 200 ml water) was added to each trial. Serum samples were collected at predetermined time points for pharmacokinetic analysis. The time to reach maximum serum concentration (Tmax) was considered as gastric emptying rate. Compared to Water 4, Tmax of 25% Na2SO4 and 25% MgSO4 was reached significantly later, the maximum serum concentration (Cmax) of acetaminophen was significantly lower and the area under the curve determined up to 90 min (AUC90) was significantly smaller. Isotonic solutions of saline cathartics (1.8% Na2SO4, 4.2% MgSO4) did not influence the gastric emptying rate. Hypertonic solutions of saline cathartics (25% Na2SO4, 25% MgSO4) significantly delayed the gastric emptying rate.

Dysphagia caused by focal guttural pouch mycosis: mononeuropathy of the pharyngeal ramus of the vagal nerve in a 20-year-old pony mare.

A 20-year-old pony mare was presented to the equine hospital with a ten-day history of dysphagia, regurgitation and coughing. An obstruction of the oesophagus was excluded via endoscopy, but the proximal oesophagus appeared to be distended and circular contractions were missing. A guttural pouch endoscopy revealed a single, black-mottled plaque on the pharyngeal ramus of the vagus nerve in the left guttural pouch, causing a local swelling of this nerve. The pharyngeal ramus seemed to be atrophic distal to the lesion. A biopsy was taken from the lesion and histopathological findings proved the reasonable suspicion of a guttural pouch mycosis with a high degree of purulent-necrotic inflammation and invasion of fungal hyphae. There were no signs of neoplasia, such as melanoma. Daily guttural pouch irrigations with a clotrimazole emulsion (20 g Canesten(®) Gyn(4) solved in 500 ml water), led to a good recovery of the mucosa above the nerve. Periodic endoscopic examination of the left guttural pouch showed that local thickening and distal atrophy of this pharyngeal ramus did not improve, neither did the clinical symptoms. Due to progressive weight loss, acute respiratory distress and aspiration pneumonia, the 20-year-old pony mare unfortunately had to be euthanized three weeks after discharge. This case report emphasizes the enormous importance of a single nerve for the realization of the swallowing process. The one-sided loss of function of the pharyngeal branch of the vagal nerve cannot be compensated neither by the remaining ipsilateral nerves nor by the contralateral normal functioning glossopharyngeal and vagal nerves and thus inevitably leads to severe dysphagia.

Influence of laxatives on gastric emptying in healthy Warmblood horses evaluated with the D-xylose absorption test.

The use of laxatives is crucial in the treatment of horses with large colon impaction. To reach the impacted mass, the laxative must leave the stomach and pass through the small intestine. The aim of this study was to determine whether the most frequently used saline and lubricant laxatives influence gastric emptying. Six fasted normal adult Warmblood horses were used in a randomized study design with five laxative trials (1.8% sodium sulfate [1.8% Na2SO4], 4.2% magnesium sulfate [4.2% MgSO4], mineral oil [MOil], 25% sodium sulfate [25% Na2SO4], 25% magnesium sulfate [25% MgSO4]) and two trials with water (at either 20 ml/kg BW [Water 20] or 4 ml/kg BW [Water 4]), administered via nasogastric intubation. For indirect measurement of liquid-phase gastric emptying, a liquid passage marker (0.5 g D-xylose/kg BW as 10% solution) was added to each trial. Serum samples were collected at pre-determined time points for pharmacokinetic analysis. The time to reach maximum serum concentration (T(max)) was considered as gastric emptying rate. Significant differences were detected for T(max) of 4.2% MgSO4 compared to Water 20 and for T(max), the maximum serum concentration (C(max)) and the area under the curve determined up to 90 min (AUC90) of 25% Na2SO4 and 25% MgSO4 compared to Water 4. Neither 1.8% Na2SO4, nor MOil delayed gastric emptying rate compared to water (Water 20, Water 4, respectively). 4.2% MgSO4 as well as 25% Na2SO4 and 25% MgSO4 significantly delayed gastric emptying rate in comparison to water (Water 20, Water 4, respectively).

A follow up study on antibodies against Lawsonia intracellularis in mares and foals from two breeding farms in Germany.

Equine proliferative enteropathy (EPE) caused by Lawsonia (L.) intracellularis is an emerging disease in foals, particularly in North America. Since a case report in Germany exists, the objective of this study was to examine the incidence of L. intracellularis-antibodies in healthy horses from two German breeding farms. In group 1, serum samples from 24 (year 1) and 16 (year 2) Haflinger mares and their foals were taken. In group 2, over a period of five months, serum samples of six warmblood mares and foals were collected monthly from birth until the foals became seronegative. Serum samples were tested using an ELISA system. Results are expressed as Percentage of Inhibiton (PI). All adult mares (100%) of both groups were seropositive at each point in time (PI-value > 30). In group 1,7/24 foals (29.2%) in year 1 and 4/16 foals (25%) in year 2 had antibodies.The seropositive foals from year 2 had the same dams as the seropositive foals from year 1. In group 2 five of six foals were seropositive after birth. Antibodies decreased from March to July in mares and foals. In July, all five foals tested negative for the first time between the ages of 82 and 141 days (median 115 days). PI-values of mares were significantly correlated with PI-values of their foals. Higher PI-values were seen in younger foals and early in spring. Loss of antibodies in foals at the age of three to five months could be a risk factor for infection and appearance of EPE.

Differentiation of viable, apoptotic and necrotic cells in bronchoalveolar lavage fluid of normal horses and horses with recurrent airway obstruction.

Proinflammatoric cytokines are released extracellularly during necrosis. These lead to inflammation and destruction of surrounding tissues. The aim of this study was to compare the number of viable, apoptotic and necrotic cells in the bronchoalveolar lavage fluid (BALF) of normal horses and horses with recurrent airway obstruction (RAO) and to determine if fluorescence microscopy is a reliable method for this examination. A group of six normal horses and a group of ten horses with RAO were examined. Samples were assessed using annexin-V and propidium iodide immunofluorescence assay and examined by fluorescence microscopy (16 horses) and flow cytometry (nine of 16 horses). We found no significant differences in percentages of apoptotic and viable cells between both groups. The number of necrotic cells was significantly increased in horses with RAO counted by fluorescence microscopy. Cells with high granularity and macrophages had a significantly higher percentage of necrotic cells than lymphocytes. There was a good agreement between both methods. No significant differences were detected. The correlation between both methods is significant. Higher amounts of necrotic cells in the bronchial lumina of horses with RAO could be a reason for tissue damage and continuous lung tissue inflammation. Fluorescence microscopy was applicable for examination of BALF. Therapy should be aimed at the reduction of necrotic cells in the bronchial lumina. Further studies are required to find ways to reduce inflammatory cell infiltration and necrosis in bronchial lumina.

Effect of Buscopan compositum on the motility of the duodenum, cecum and left ventral colon in healthy conscious horses.

Seven adult, healthy, conscious warmblood horses were used in a crossover study. They were fed twice a day on 1 kg hay/100 kg BW and 0.5 kg concentrates with unlimited access to water. One hour after feeding, the contractive motility of the descending duodenum, cecal body and left ventral colon were measured using a 5 MHz transcutaneus ultrasonographic transducer. Each horse was treated with 0.9% NaCl (5 ml/100 kg BW; i.v.), and with Buscopan compositum (BC) at its therapeutic dosage (25 mg/kg BW; metamizol-sodium, 0.2 mg/kg BW; N-butylscopolammonium bromide, i.v.) in a control and an experimental trial respectively. The contractive motility was counted in a 3-min period immediately and 5, 15, 30, 45, 60, 120, 180 and 240 min after application in both trials. There was an immediate, rapid and significant (p < 0.05) reduction of duodenal, cecal and left ventral colon contractions after BC administration. Cecal and left ventral colon contractions rapidly restored their normal contractions after 30 min, while duodenal contractions returned to the normal rate after 120 min of BC administration. It was concluded that Buscopan compositum at its therapeutic dosage has an immediate, potent, short-lived reductive effect on cecum and left ventral colon contractions but a minor, longer effect on the duodenal contractions. Therefore, it is recommended for treatment of spasmodic colic as well as spasms resulting from acute simple impaction in horses but it is not recommended for repeated short-interval administration.

[Free hemoglobin and tumor necrosis factor-alpha in the blood of horses with colic or acute colitis]. / Freies Hämoglobin und Tumor-Nekrose-Faktor-alpha im Blut von Pferden mit Kolik oder akuter Kolitis.

A total of 50 adult horses were classified into 4 groups: healthy horses (group 1, n = 14), horses with non-strangulation obstruction (group 2, n = 13), horses with strangulation obstruction (group 3, n = 11) and horses with acute colitis and severe inflammation (peritonitis or thrombophlebitis) (group 4, n = 12). Eleven euthanized horses recreated from group 3 (4 horses with small intestinal strangulation obstruction and hemolytic peritoneal fluid) and group 4 (7 horses with septic peritonitis) were taken as group 5. Free hemoglobin (fr. Hb) and tumor necrosis factor-alpha (TNFa) were measured by method of Kahn and by Duo Set ELISA Development Kit. The concentrations of fr. Hb and TNF-a were significantly higher (p <0.01) in group 3 (medians: 8.0 micromol/l; 14665 pg/ml) and group 4 (9.1; 15081) in comparison to healthy horses (0.61; 573) and group 2 (0.5; 821), whereas group 2 showed no significant difference with group 1.There was a positive correlation between fr. Hb and TNF-alpha (r = 0.82; P = 0.01) in group 4. Concentrations of both fr. Hb and TNF-alpha were significantly (p <0,01) higher in euthanized horses of group 5 than in medically treated horses of group 2.The high concentration of fr. Hb could be a proinflammatory mediator to stimulate high production ofTNF-alpha known as a proinflammatory cytokine. Both of these parameters could contribute to the pathogenesis of systemic inflammatory response syndrome: fever, tachycardia, tachypnoe, leukopenia, thrombocytopenia, and lactic acidosis. Based on this, increased concentrations of both fr. Hb and TNF-alpha could be a guarded prognostic indicator in horses with strangulation obstruction, acute colitis or septic peritonitis.