Impact of spontaneous donor hypothermia on graft outcomes after kidney transplantation.

A previous donor intervention trial found that therapeutic hypothermia reduced delayed graft function (DGF) after kidney transplantation. This retrospective cohort study nested in the randomized dopamine trial (ClinicalTrials.gov identifier: NCT000115115) investigates the effects of spontaneous donor hypothermia (core body temperature <36°C) on initial kidney graft function, and evaluates 5-year graft survival. Hypothermia assessed by a singular measurement in the intensive care unit 4-20 hours before procurement was associated with less DGF after kidney transplantation (odds ratio [OR] 0.56, 95% confidence interval [CI] 0.34-0.91). The benefit was greater when need for more than a single posttransplant dialysis session was analyzed (OR 0.48, 95%CI 0.28-0.82). Donor dopamine ameliorated dialysis requirement independently from hypothermia in a temporal relationship with exposure (OR 0.93, 95%CI 0.87-0.98, per hour). A lower core body temperature in the donor was associated with lower serum creatinine levels before procurement, which may reflect lower systemic inflammation and attenuated renal injury from brain death. Despite a considerable effect on DGF, our study failed to demonstrate a graft survival advantage (hazard ratio [HR] 0.83, 95%CI 0.54-1.27), whereas dopamine treatment was associated with improved long-term outcome (HR 0.95, 95%CI 0.91-0.99 per hour).

Analysis of gene expression in granulosa cells post-maturation to evaluate oocyte culture systems in the domestic cat.

Maturation of oocytes is a prerequisite for successful embryo development. The fertilization competence of in vivo derived oocytes is significantly higher than that of oocytes matured in vitro. Commonly evaluated morphological criteria for oocyte maturation do not reflect the complexity and quality of maturation processes. Oocytes and granulosa cells are communicating closely in a bidirectional way during follicular growth and maturation. Assessing the mRNA expression of specific genes in granulosa cells could be a non-invasive way to evaluate the conditions of in vitro oocyte maturation. The objective of this study was to elucidate the influence of two different FSH additives on the in vitro maturation rate and gene expression of cumulus-oocytes complexes in domestic cat. Feline oocytes were matured in a medium, supplemented with LH and 0.02 IU/ml porcine FSH versus 0.02 IU or 1.06 IU/ml human FSH. Granulosa cells were separated from oocytes directly after 24 hr of maturation or after additional 12 hr of in vitro fertilization. Gene expression levels were analysed by quantitative PCR for aromatase, antimullerian hormone, follicle stimulating hormone receptor (FSHR), luteinizing hormone/choriogonadotropin receptor (LHCGR) and prostaglandin E synthase. Neither oocyte maturation rate nor gene expression levels differed after 24 or 36 hr in all three groups. However, variations were discovered in correlations of expression levels, for instance for FSHR and LHCG, indicating differences in the fine-tuning of in vitro maturation processes under varying FSH supplementations. We suppose that correlation between gene expressions of selected genes suggests a superior maturation quality of feline oocytes.

Efficacy of toltrazuril (Baycox 5% suspension) in natural infections with pathogenic Eimeria spp. in housed lambs.

A field study was conducted to evaluate the effect of a single oral treatment with 20 mg/kg body weight (BW) of toltrazuril (Baycox 5% suspension)--TOL--in comparison to a single oral treatment with 1 mg/kg BW of diclazuril (Vecoxan suspension orale, 2.5 mg/ml)--DIC--and an untreated control group (CTRL) on naturally acquired Eimeria infections in lambs. On a French sheep farm with a known history of coccidiosis, 75 housed lambs aged 10-14 days were randomised and allocated to one of three groups. During an observation period of 60 days after treatment, clinical (faecal consistency, BW) and parasitological parameters (oocyst excretion) were evaluated. Excretion in the negative control group started 3 days after treatment and peaked on the 31st day with a prevalence of 80%. Animals were predominantly infected with Eimeria ovinoidalis. Treatment with toltrazuril, but not with diclazuril, resulted in significantly reduced numbers of excreting animals. The number of excretion days and the average oocyst excretion decreased significantly in both the TOL and the DIC groups compared to the CTRL, with the TOL group showing significantly fewer excretion days and excretion intensities than the DIC group. Changes in the faecal consistency were moderate throughout the study and not significantly different between the groups. Daily weight gains were higher in the TOL group compared to the DIC and CTRL groups which did not differ. This study demonstrates the good efficacy of toltrazuril administered orally to lambs in the prepatent period in subclinical natural Eimeria infections in housed lambs.

Efficacy of various anticoccidials against experimental porcine neonatal isosporosis.

The efficacies of 20 mg/kg body weight (BW) of toltrazuril (Toltra) 2 days post-infection (dpi), 2 mg/kg BW of diclazuril 2 and 3 dpi and 200 mg/kg BW of sulphadimidine 2, 3 and 4 dpi were compared in a model for piglet isosporosis. Weight gain (first 4 weeks of life) and diarrhoea and oocyst excretion from 4 to 11 dpi were evaluated (10-12 piglets/group). Additionally, animals were killed and examined for pathohistological changes of the small intestines 5, 7, 11 and 14 dpi (n = 3 per group and time point) and lengths of the intestinal villi. Diarrhoea (semi-liquid or liquid faeces) was seen from 5 dpi in all groups except Toltra, and the differences in prevalence and intensity of diarrhoea were statistically significant (p < 0.05) between Toltra and the other groups, which were similar (trial 1). Oocyst excretion was greatly reduced in the Toltra group, which was also statistically significant for the mean and median excretion rates and the percentage of excreting piglets between Toltra and the other groups (p < 0.05). Weight gain was highest in Toltra (p < 0.05). Histopathology revealed mostly villous necrosis and atrophy in the small intestines except the duodenum, which peaked at 7 dpi, in all groups except Toltra. Between 5 and 11 dpi, the Toltra group had significantly longer villi than the other groups. Reduced weight gain and diarrhoea caused by Isospora suis was controlled by a single application of Toltra in the pre-patent period, while neither diclazuril nor sulphadimidine improved the clinical picture of isosporosis.

Canine isosporosis - epidemiology of field and experimental infections.

Isospora spp. are the causative agents of canine isosporosis. Of the 3590 diagnostic samples from Austrian dogs (< or = 2 years old), 8.7% contained Isospora oocysts, 78% of which from dogs up to 4 months of age. Non-haemorrhagic and haemorrhagic diarrhoea were significantly more prevalent in Isospora-infected animals than in coccidia-negative ones. Twelve of 15 litters from a large commercial dog breeding unit (examined from the third to the 10th week of life) also excreted Isospora (average prevalence: 36.4%) in intensities from 333 to 35,000 oocysts per gram of faeces (opg). In experimental trials 26 3-week-old Beagle puppies were infected with low (600-6000), medium (10,000) or high (20,000) dose of Isospora ohioensis-group or Isospora canis field isolates. Additionally 21 puppies were infected as above and treated with a symmetrical triazintrione. Parasitological and clinical parameters were examined. The two Isospora species differed significantly concerning intensity and duration of excretion. The pre-patent period was 6-7 days for I. ohioensis and 10-12 days for I. canis. The latter species showed significantly longer excretion and higher opg. This was not influenced by simultaneous infections with both species. Individual patterns of faecal consistency were very variable, irrespective of the infection dose. Treatment significantly reduced both the intensity and the duration of oocysts excretion as well as diarrhoea in comparison with the infected, untreated group and thus proved to be effective against coccidiosis in experimental infections.

Isospora suis: an experimental model for mammalian intestinal coccidiosis.

Piglets experimentally infected with 10,000 oocysts of Isospora suis in three identical trials (n = 50) were examined clinically and coproscopically from 5 to 11 days post-infection (d.p.i.), weighed in weekly intervals until the fourth week of life and compared to age-matched asymptomatic controls (n = 17). Furthermore, 17 infected piglets were histologically examined on days 5-14 p.i. Infected animals had a significantly lower weight gain than the controls and showed diarrhoea throughout, with maximum prevalence and intensity on 6 d.p.i. Half of the animals had diarrhoea for only 2 days or less. The number of diarrhoea days was negatively correlated with weight gain. Oocyst excretion started on 5 d.p.i. with peak prevalences and declined afterwards; a smaller peak was seen on 10 d.p.i. All animals excreted parasites at least once, and most of them excreted for 5-7 days. Oocyst excretion intensity paralleled the prevalence and ranged from 220 to 251,501 oocysts per gram of faeces (opg). Most samples contained 4 x 10(3) to 4 x 10(4) opg. The opg values were negatively correlated with faecal scores (samples with diarrhoea contained less oocysts) of the same day and the previous day. Histologically, necrosis followed by atrophy of the villi was most pronounced in the early stage of infection throughout the jejunum and ileum but declined thereafter. On 14 d.p.i., villous atrophy was still noticeable in the jejunum. Histology is difficult to quantify and requires large animal numbers, although the effects are visible for some time. Weight gain and faecal score can be affected by other factors than parasite infection. From the compiled data, we conclude that the established model is suitable to study piglet isosporosis with oocyst excretion being the most reliable parameter, although individual variations are considerable. A negative correlation between excretion and diarrhoea may be responsible for the difficulties in the detection of the parasite in field samples.

Control of clinical coccidiosis of calves due to Eimeria bovis and Eimeria zuernii with toltrazuril under field conditions.

The efficacy of metaphylactic treatment with toltrazuril (Baycox 5% suspension) against natural infections with Eimeria bovis and/or Eimeria zuernii in calves was investigated. The study was conducted with 208 calves on five calf-rearing farms in Germany and the Czech Republic. All participating farms had a notable incidence of coccidiosis. Animals were treated 14 days after stabling in the respective facility. One group was treated with 15 mg toltrazuril/kg body weight, and a second group served as the sham-treated control. Assessment of efficacy was based on faecal consistency and oocyst excretion of E. bovis and E. zuernii, both investigated throughout the study. Duration and rate of oocyst excretion as well as number of scour days with E. bovis or E. zuernii oocyst shedding and the severity of diarrhoea were significantly lower in the toltrazuril-treated groups. It is concluded that a single metaphylactic treatment with toltrazuril controls coccidiosis of housed calves under various field conditions.

Pathology and treatment of Eimeria zuernii coccidiosis in calves: investigations in an infection model.

Two studies were conducted in the Eimeria zuernii infection model in order to investigate the pathology of E. zuernii coccidiosis and the efficacy of toltrazuril (Baycox 5% suspension) in this infection. For this purpose, a total of 30 calves were infected experimentally with E. zuernii oocysts and faecal samples taken regularly from the rectum and examined for faecal consistency and oocyst excretion. Six of the calves underwent pathological examination at various points in time after infection. Significant macroscopic and microscopic changes were demonstrated and parasitic stages were identified in the intestinal mucosa of infected calves during the late prepatent and patent period. Inflammatory reactions revealed by light microscopy were confirmed by electron microscopical investigations. Treatment of calves with toltrazuril during the late prepatent period resulted in significantly lower frequencies of diarrhoea and levels of oocyst excretion, and weight gain was significantly higher than in shamtreated animals.

Occurrence of Isospora suis in Germany, Switzerland and Austria.

Nationwide surveys for the occurrence of Isospora suis were carried out in Germany, Austria and Switzerland including a questionnaire regarding herd size, health status and management practices and a coccidiosis sampling kit for pooled faecal samples from litters of suckling piglets. A total of 184 veterinary practices participated in the survey and returned 1745 samples (331 kits) from 324 farms in the north (n = 98), south (n = 84), centre/east (n = 42) and west (n = 10) of Germany, Austria (n = 61) and Switzerland (n = 29) with larger farms in north and centre/east (average number of sows: 270 and 500) and smaller ones in the south (95), Austria (60) and Switzerland (43). Larger farms tended to have better hygienic standards (slatted floors, disinfection of the farrowing units). The majority of the participating farms (93.5%) reported problems with diarrhoea in piglets at 2-3 weeks of age, significantly associated (P < 0.001) with uneven weaning weights (94.9%). Toltrazuril (5%; Baycox) was used only rarely; however, in these farms unevenness of weaning weights was less frequently observed (P = 0.011). A 76.2% of the farms were positive for I. suis (samples contained mostly low or moderate oocyst numbers), especially in the south (P < 0.001). Oocysts were more frequently found in samples from farms with reported diarrhoea (P = 0.011), uneven weight gain (P = 0.019) or in herds of small size (P < 0.001). Disinfection, floor type or treatment with toltrazuril did not affect the frequency of observation of oocysts.

World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines for evaluating the efficacy of anticoccidial drugs in chickens and turkeys.

These guidelines have been written to aid in the design, implementation and interpretation of studies for the assessment of drug efficacy against Eimeria species in chickens and turkeys. The information provided deals with many aspects of how to conduct controlled studies in battery cages (dose determination), floor pens (dose confirmation), and commercial facilities (field effectiveness studies), the selection of birds, housing, feeding, preparation of medicated rations, record keeping, diagnostic techniques, and methods for the preparation, maintenance and use of parasites. These guidelines are also intended to assist investigators in conducting specific studies, provide specific information for registration authorities involved in the decision-making process, assist in the approval and registration of new anticoccidial drugs, and facilitate the world-wide adoption of standard procedures.

Autofluorescence microscopy for the detection of nematode eggs and protozoa, in particular Isospora suis, in swine faeces.

Parasites from swine faeces were examined for autofluorescence. Oocysts of Eimeria polita, E. scabra and Isospora suis, cysts of Balantidium coli and eggs of Oesophagostomum dentatum, Strongyloides ransomi and Trichuris suis (but not those of Ascaris suum) emitted light after excitation with UV light. I. suis oocyst counts in McMaster chambers utilising autofluorescence were compared to those from conventional bright field microscopy. Similarly, faecal smears containing I. suis were examined using the same techniques. Autofluorescence was superior to bright field microscopy in detecting oocysts after flotation and was highly significantly more sensitive when direct smears were examined.

Toltrazuril treatment of cystoisosporosis in dogs under experimental and field conditions.

Coccidia of the genus Cystoisospora cause mild to severe diarrhoea in dogs. The effects of toltrazuril treatment on cystoisosporosis were studied under experimental and field conditions. Twenty-four puppies were experimentally infected each with 4 x 10(4) oocysts of the Cystoisospora ohioensis group. Three groups of six puppies were treated 3 dpi with 10, 20 or 30 mg/kg body weight of toltrazuril suspension (5%); the remaining six puppies served as non-treated controls. Toltrazuril suspension or microgranulate were given once in a dose of 10 or 20 mg/kg body weight, respectively, to naturally infected puppies in conventional dog breeding facilities, depending on the coproscopical evidence of infection. Oocyst excretion and clinical data were recorded. Under experimental conditions, the non-treated puppies excreted oocysts beginning at 6 dpi and suffered from catarrhalic to haemorrhagic diarrhoea. On 12 dpi, four of six non-treated puppies died. Irrespective of the dose, toltrazuril treatment totally suppressed oocyst excretion and no diarrhoea or other signs of disease were observed in the treated groups. Natural Cystoisospora infections were regularly found during the 3rd or 4th week of age in dog breeding facilities although not always associated with diarrhoea. A single oral application of toltrazuril abrogated oocyst shedding and the treated puppies remained generally coproscopically negative during the following 2-4 weeks. Cystoisospora is pathogenic for puppies and can induce severe disease. Natural infections are common in conventional dog breeding facilities. Toltrazuril treatment is suitable for controlling cystoisosporosis under experimental and field conditions. A single oral treatment for puppies in the 3rd or 4th week of age is recommended.

[Sulfonamide and toltrazuril therapy of experimental turkey coccidiosis]. / Zur Sulfonamid- und Toltrazuriltherapie der experimentellen Putenkokzidiose.

Following experimental infection of caged turkeys with a mixed oocyst suspension of E. meleagrimitis, E. adenoeides and E. gallopavonis, the efficacy of sulfachlorpyrazine, sulfaquinoxaline + pyrimethamine, sulfaquinoxaline + diaveridine, and toltrazuril was investigated based on clinical and parasitological parameters. Infected turkeys developed severe clinical symptoms. Only weak symptoms were observed in those groups which were treated with the sulfa- and sulfa-DHFR-inhibitors. The toltrazuril-treated group remained free of clinical symptoms. At the 8th day after infection, the clinical lesions coincided with the intensity of the gut lesions together with parasitological findings based on microscopical examination of mucosal smears. The same coincidence was observed for parameters such as oocyst excretion and body weight development.

[Use of toltrazuril in pullet breeding flocks raised on floors with anticoccidial-free feed]. / Einsatz von Toltrazuril bei Junghennenaufzuchten in Bodenhaltung mit antikokzidiafreier Fütterung.

In 28 replacement pullet flocks with flock size of 4,000 to 25,000 birds Toltrazuril (experimental preparation, 2.5% solution, approx. 7 mg/kg body weight) was administered for a period of two days in the drinking water for metaphylactic purpose and therapeutic use, respectively. All flocks were reared without anticoccidial feed additives. Exposition to Eimeria spp. was recorded by examination of faecal samples and intestinal scrapings. Clinical flock control was achieved by vets at eight to ten day intervals. Toltrazuril was characterized by short application period, rapid mode of action, significant reduction of oocyst shedding, good compatibility and acceptance thus proving superior to conventional chemotherapy of coccidiosis. The results suggest that, even without administration of anticoccidial feed additives. Toltrazuril is an effective agent for therapeutic use in acute coccidiosis. Under insufficient management and rearing conditions repeated metaphylactic application has to be considered.