Analysis of potential risk factors of caprine coccidiosis.

Caprine coccidiosis represents a major concern in goat industry worldwide causing high economic losses, mainly due to poor weight gains, treatment costs and even death of kids. Therefore, analyses on risk factors involved in the establishment and outcome of this disease are important to improve infection status and herd management. In Portugal, approximately 400,000 goats are reared every year. The majority of them are allocated in the southern part of the country. In the current study, a representative number of animals (382 goats) and herds (37 herds) from Alentejo were monitored. Faecal samples from five young and five adult goats of each farm were collected rectally and examined individually by using the modified McMaster technique for oocyst counts (oocyst per gram of faeces - OPG). Additionally, a questionnaire on the husbandry conditions and anti-parasitic treatments was performed. Overall, a prevalence of Eimeria infections of 99% and 100% was detected on animal and herd level, respectively. Eimeria arloingi and E. ninakohlyakimovae as the two most pathogenic caprine species also constituted the most prevalent ones (34.91% and 29.90%, respectively), both in the young and the adult goat groups. As expected, OPG vales were significantly higher (p = 0.031) in young than in adult goats. Furthermore, risk factor analyses revealed young age, higher total number of animals on the farm and outdoor kidding as strongly associated with a higher risk of caprine coccidiosis. Overall, the high prevalence of pathogenic Eimeria species on Portuguese goat farms in combination with the occurrence of related risk factors strengthen the importance of coccidiosis control in caprine industry.

Epidemiological survey and risk factor analysis on Eimeria infections in calves and young cattle up to 1 year old in Colombia.

A large-scale cross-sectional epidemiological study was conducted to evaluate prevalence, species diversity, and associated risk factors of Eimeria infections in 55 cattle farms across seven states of Colombia, including subtropical and tropical regions. In total, 1333 fecal samples from young animals (< 1 year of age) were examined at a single sampling date from August 2016 to December 2016. Flotation and McMaster techniques were conducted for parasitological investigation. Excreted Eimeria oocysts were allowed to sporulate in vitro and thereafter identified to species level based on morphological and morphometric characteristics. The overall Eimeria prevalence was 75.5% (1006/1333), with no difference observed between age categories. In total, 13 different Eimeria species were identified. The most prevalent species was E. bovis (33.5%), followed by E. auburnensis (12.5%) and E. zuernii (11.9%). Analysis of extrinsic associated risk factors revealed the floor type, feeding system, watering system, and herd size as significant (p < 0.05) risk factors for Eimeria spp. infections. Based on these data, it can be assumed that bovine coccidiosis infections occur ubiquitously in the country and might play an important role especially in its subclinical form by affecting production parameters in conventional cattle management systems.

Automated detection of health disorders in lactating dairy cattle on pasture: a preliminary study.

Since previous health monitoring systems have shown themselves to be unsuccessful in predicting health disorders in dairy cows managed on pasture, the aim of this study was to evaluate the performance of automated health monitoring integrated in an accelerometer-based oestrus detection system (ODS) for dairy cows on pasture. Mixed-breed lactating dairy cows (n=109) in a seasonal-calving herd managed at pasture were fitted with an ODS that provided automated health monitoring. The ODS performed multimetric analysis of behavioural patterns to generate health alerts. Data were collected during the artificial insemination period of 66 days. Clinical examinations and farmer's observations were used to evaluate the performance of automated health monitoring. During the insemination period, the farmer generated two health alerts, which were classified false positives (2/2; 100%). The ODS generated 31 automated health alerts. Of all automated health alerts, 3/31 (9.7%) were confirmed as true health disorders and 28/31 (90.3%) alerts were classified as false positives. The positive predictive value (PPV) of automated health monitoring was 9.7 (95% CI=2-25.8) %. The ODS was able to alert lactating dairy cows on pasture suffering from health disorders. True health disorders were alerted by the ODS before the farmer noticed them, which could provide early and successful treatment when using the system on-farm for automated health monitoring. The evaluated accuracy of automated health monitoring is opposed to a targeted use of the system for on-farm health monitoring. For further validation, testing on other farms and during the transition period would be of interest.

Semen collection, semen analysis and artificial insemination in Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus) as part of a species conservation project.

Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus; hereafter CSTG) have experienced substantial decreases in population numbers and geographic range during the early 20th century, primarily due to habitat loss. The conservation aim of this project was to re-establish a self-sustaining population of CSTG within an unoccupied portion of their historic range in northeastern Nevada via reintroduction from source populations in Idaho, USA. Female nest initiation rates post-translocation due to low fertilization rates are believed to be one limiting factor in the establishment of some translocated CSTG populations. However, studies on semen collection and artificial insemination in this species are absent. Assisted reproduction was evaluated as an additional tool in this species conservation project in order to gain knowledge on the reproductive status of yearling and adult male CSTG, establish orientation values for semen parameters and evaluate artificial insemination procedures on female CSTG. In two consecutive breeding seasons, semen collection was attempted 51 times in 47 males using the established massage method, and a novel electro-stimulation technique. Semen collection was successful in all attempts, even in yearling grouse, which represents a novel confirmation that yearling male CSTG can produce live spermatozoa in their first breeding season. Volume, color, consistency, contamination, pH of semen, and the motility, concentration, viability and morphology of spermatozoa were analyzed. Extracted semen volume ranged between 6 and 74 µl and the mean pH was 6.9 ±â€¯0.5 (x¯ ± SD). Morphology analysis revealed that on average, 42.8% of sperm was morphologically normal, but 34.8% had malformed tails. Additionally, artificial insemination was practiced in 17 females (sham-insemination group; insemination lacking spermatozoa) and performed in 17 females. Intravenous catheters G20 1.0 mm × 32 mm enabled safe intravaginal insemination under visual control. Significant (p ≤ 0.05) differences in semen parameters between adult and yearling birds were detected. It is well established that adult males receive a majority of copulations during lekking, but our novel findings also indicate that they produce significantly more spermatozoa which is of higher quality than yearling males. This finding offers insights into male reproductive biology in a lekking grouse species.

Comparative ultrasonographic examination and measurements of the urethra and penis of castrated and intact male lambs.

Early castration of male small ruminants is regarded as a risk factor for urolithiasis, although the underlying correlations are still unclear. One possible reason is a deferred development of the penis and the urethra after castration. Therefore, we examined the penis and urethra of castrated and intact lambs by ultrasonography to determine the correlation between urethral area and pe- nile cross-sectional area. Ultrasonography was performed in 6-month-old Lacaune crossbred lambs (early castrated, late castrated, and intact; each group, n = 11). Sectional images at 5 loca- tions (glans penis, penile urethra, distal and proximal sigmoid flexure, and ischial arch) were ob- tained to determine the urethral and penile diameters. Urethral and penile cross-sectional areas were calculated. Grey-scale analysis of ultrasound images was performed to evaluate possible differences in the penile texture between the groups. Correlation analyses between both cross-sectional areas showed a significant general correlation for location 2 in all lambs (R = 0.52; P = 0.003), for location 3 in late-castrated lambs, and for location 5 in early-castrated lambs. Statistically significant correlations between the penile and the urethral area of castrated and intact lambs were not evident. Therefore, measurement of the penile cross-sectional area alone does not allow for accurate estimation of urethral size. Statistically significant differences con- cerning the grey-scale analysis between the groups were also not detectable. Thus, simplification of the formerly presented ultrasonographic examination of the urethra is not recommended. In animals at a risk of obstructive urolithiasis, complete urethral examina- tion is essential.

The role of Virus "X" (Tortoise Picornavirus) in kidney disease and shell weakness syndrome in European tortoise species determined by experimental infection.

Tortoise Picornavirus (ToPV) commonly known as Virus "X" was recently discovered in juvenile European tortoises suffering from soft carapace and plastron as well as kidney disease. Therefore, this virus was a potential candidate to be a causative agent for these disease patterns. Spur thighed tortoises (Testudo graeca) seemed to be more susceptible to establish clinical symptoms than other European species like T. hermanni. Thus this trial investigated the role of ToPV in the described syndrome. Two groups of juvenile European tortoises (T. graeca and T.hermanni) each of 10 animals, were cloacally, oronasally and intracoelomically inoculated with an infectious dose (~ 2000 TICD) of a ToPV strain isolated from a diseased T. graeca. A control group of two animals of each species received non-infected cell culture supernatant. The tortoises were examined daily and pharyngeal and cloacal swabs for detection of ToPV-RNA by RT-PCR were taken from each animal every six days for a period of 6 months. At the end of the study the remaining animals were euthanised and dissected. Bacteriological and parasitological tests were performed and organ samples of all tortoises were investigated by RT-PCR for the presence of ToPV and histopathology. Animals that were euthanised at the end of the experiment, were examined for presence of specific anti-ToPV antibodies. Several animals in both inoculated groups showed retarded growth and a light shell weakness, in comparison to the control animals. Three animals were euthanised during the trial, showing reduced weight gain, retarded growth, severe shell weakness and apathy, in parallel to clinical observations in naturally infected animals. In all inoculated animals of both species an intermittent virus shedding, starting from 18 days post inoculation (d.p.i.), till 164 d.p.i. was detected, while the control animals remained negative. The virus was successfully reisolated in terrapene heart cell culture in 16 of 20 inoculated animals of both species. Histopathology of most inoculated animals revealed a lack of bone remodeling and vacuolisation in kidney tubuli which supports the described pathogenesis of nephropathy and osteodystrophy. Anti- ToPV antibody titres ranged from 1:2 to >1:256 in 13 of 20 animals, whereas all control animals were seronegative. The study proofed the Henle Koch`s postulates of ToPV as causative agent for shell dystrophy and kidney disease in both testudo species. The proposed species specific sensitivity towards clinical disease was not observed.

Associated risk factors influencing ovine Eimeria infections in southern Spain.

Eimeria infections are common in sheep industry worldwide nonetheless very little is known on infection dynamics and associated risk factors influencing the outcome of clinical ovine coccidiosis. The aim of the present work was to analyse intrinsic (e. g. host age) and extrinsic (flock size, farm size and animal density) risk factors influencing dynamics of Eimeria spp. infections in sheep farms of the semi-arid region of Murcia in southern Spain. Overall, 64 ovine flocks in Murcia were surveyed and a total of 396 faecal samples were collected between May and July 2015. Flotation and McMaster techniques were performed for parasitological investigation. Excreted oocysts were identified on the basis of morphological as well as morphometric characteristics. In all analysed samples Eimeria spp. oocysts (100%) were found and 11 different Eimeria species were identified. Most prevalent species was E. ovinoidalis (97%) followed by E. parva/E.pallida (97%), E. weybridgensis (90.9%) and E. crandallis (89.4%), respectively. Concomitant infections with more than one Eimeria species were observed in all analysed samples. Analysis of intrinsic/extrinsic risk factor revealed age, herd/farm size and animal density to be the main factors significantly influencing Eimeria oocysts per gram of faeces counts (OPG) and overall prevalence of Eimeria spp. infections in sheep.

Functional anatomy of the equine temporomandibular joint: Histological characteristics of the articular surfaces and underlining tissues.

It has been assumed that dental conditions cause disorders of the equine temporomandibular joint (TMJ), due to biomechanical overload or aberrant loading. However, the incidence of published TMJ disorders in horses is low and this leads to the question whether the equine TMJ is adapted well to its biomechanical requirements or is able to remodel its articular surfaces in response to modified loading conditions. The aim of this study was to determine the histological characteristics of healthy equine TMJs. The tissue components of the articular surfaces of 10 TMJs obtained from horses without any clinical history of dental or TMJ disorders were analysed. Apart from the mandibular fossa of the temporal bone, the osseous aspects of the TMJ exhibited a uniform zoning pattern. The articular surfaces were composed of three tissue layers: (1) a superficial cell-rich dense connective tissue layer; (2) a middle fibrocartilage layer; and (3) a deep hyaline-like cartilage layer. The articular disc was composed of an inner core of fibrocartilage and hyaline-like cartilage meshwork covered with both cell-rich dense connective tissue and fibrocartilage at its dorsal and ventral aspects. In contrast, the mandibular fossa was only covered by a dense connective tissue, frequently supplemented by a synovial membrane, suggesting low biomechanical stress. Glycosaminoglycans, which are indicative of compressive loads, were predominantly present within the rostral part of the articular tubercle and the retroarticular process, the dorsal part of articular disc and the entire mandibular head, but were absent within the mandibular fossa. The results of this study suggest the presence of different biomechanical demands in the dorsal and ventral compartment of the equine TMJ.

Automated oestrus detection using multimetric behaviour recognition in seasonal-calving dairy cattle on pasture.

AIM To evaluate the performance of a novel accelerometer-based oestrus detection system (ODS) for dairy cows on pasture, in comparison with measurement of concentrations of progesterone in milk, ultrasonographic examination of ovaries and farmer observations. METHODS Mixed-breed lactating dairy cows (n=109) in a commercial, seasonal-calving herd managed at pasture under typical farming conditions in Ireland, were fitted with oestrus detection collars 3 weeks prior to mating start date. The ODS performed multimetric analysis of eight different motion patterns to generate oestrus alerts. Data were collected during the artificial insemination period of 66 days, commencing on 16 April 2015. Transrectal ultrasonographic examinations of the reproductive tract and measurements of concentrations of progesterone in milk were used to confirm oestrus events. Visual observations by the farmer and the number of theoretically expected oestrus events were used to evaluate the number of false negative ODS alerts. The percentage of eligible cows that were detected in oestrus at least once (and were confirmed true positives) was calculated for the first 21, 42 and 63 days of the insemination period. RESULTS During the insemination period, the ODS generated 194 oestrus alerts and 140 (72.2%) were confirmed as true positives. Six confirmed oestrus events recognised by the farmer did not generate ODS alerts. The positive predictive value of the ODS was 72.2 (95% CI=65.3-78.4)%. To account for oestrus events not identified by the ODS or the farmer, four theoretical missed oestrus events were added to the false negatives. Estimated sensitivity of the automated ODS was 93.3 (95% CI=88.1-96.8)%. The proportion of eligible cows that were detected in oestrus during the first 21 days of the insemination period was 92/106 (86.8%), and during the first 42 and 63 days of the insemination period was 103/106 (97.2%) and 105/106 (99.1%), respectively. CONCLUSIONS and CLINICAL RELEVANCE The ODS under investigation was suitable for oestrus detection in dairy cows on pasture and showed a high sensitivity of oestrus detection. Multimetric analysis of behavioural data seems to be the superior approach to developing and improving ODS for dairy cows on pasture. Due to a high proportion of false positive alerts, its use as a stand-alone system for oestrus detection cannot be recommended. As it is the first time the system was investigated, testing on other farms would be necessary for further validation.

Prevalence of Angiostrongylus vasorum, Aelurostrongylus abstrusus and Crenosoma vulpis larvae in native slug populations in Germany.

Metastrongyloid parasites represent sparsely studied parasites of dogs and cats in Germany. Recent European surveys indicate that these parasites are spreading in Europe. Actual data on prevalence of Angiostrongylus vasorum in dogs and foxes reveal several endemic foci in Germany. However, actual data on the prevalence of A. vasorum and other metastrongyloid lungworm larvae in a wide range of slug and snail intermediate hosts, such as Arion lusitanicus, are missing for Germany. To fill this gap, we conducted an epidemiological survey on native German slugs in selected regions of Hesse and Rhineland-Palatinate. The focus was on slugs, because in study areas slugs appear to be more abundant than snails. Slugs were collected throughout different seasons of the year in areas that were previously proven to be hyperendemic for A. vasorum fox infections. Overall, a total of 2701 slugs were collected and examined for lungworm larvae via artificial digestion. The number of A. vasorum larvae per slug varied considerably (1-546 larvae per specimen). Some hotspot areas with high A. vasorum prevalence in slugs (up to 19.4%) were identified. The overall A. vasorum prevalence varied with season with largest number of slugs infected in summer (9.1%) and lowest number in winter (0.8%). The current study revealed a total A. vasorum prevalence of 4.7% in slugs based on microscopic analyses. Confirmation of lungworm species was made by specific duplex-real-time PCRs. Hence, these data demonstrate that final hosts are at a permanent risk for A. vasorum infections during all seasons when living in investigated areas. Besides A. vasorum, other lungworm larvae were also detected, such as Crenosoma vulpis (the fox lungworm, 2.3%) and Aelurostrongylus abstrusus (feline lungworm, 0.2%).

Investigations on a cryopreservation protocol for long-term storage of psittacine spermatozoa using cockatiel semen as an example.

The aim of the present study was the establishment of an effective protocol for cryopreservation of psittacine semen. Therefore, pooled semen samples of 30 cockatiels (Nymphicus hollandicus) were diluted with modified Lake diluent (1:4), partitioned into four equal parts. Three portions were mixed with three cryoprotectants (dimethylacetamide, dimethyl sulfoxide, glycerol) in 4%, 8% and 12% final concentration, respectively, whereas the 4rth part served as control. Altogether, 96 incremental diluted semen samples were obtained for investigation. Each cryoprotective agent (CPA) in each final concentration was evaluated regarding sperm motility immediately after dilution and another four times every 30 min. Sperm viability was evaluated 0 and 120 min after dilution using the fluorescence stain SYBR® Green/propidium iodide. Sperm morphology was evaluated 0 and 120 min after dilution using eosin B stains. Glycerol demonstrated a lethal effect on cockatiel spermatozoa in all concentrations, whereas dimethylacetamide (DMA) in 8% final concentration proved to have the least adverse effect on semen parameters. Comparison of quick and slow freezing methods using DMA 8% revealed significantly higher rates of viable and motile spermatozoa after computer-controlled rate freezing. Two insemination experiments resulted in an egg fertility rate of 92.59% and 67.65% after artificial insemination with freshly collected semen samples, compared to 30.77% and 18.00% egg fertility rates using frozen/thawed semen. Altogether, 12 chicks hatched out of eggs inseminated with cryopreserved semen. To our knowledge, this is the first time for cockatiels to be successfully reproduced after artificial insemination using cryopreserved semen.

Influence of dental materials on cells of the equine periodontium.

BACKGROUND: Therapy for equine periodontal disease can include filling of the periodontal pockets and widened interproximal spaces. Recommended dental materials are generally adopted from human dentistry. OBJECTIVES: To evaluate the biocompatibility of dental materials for equine periodontal fillings in vitro. STUDY DESIGN: In vitro experiments. METHODS: Four different dental materials (PeriCare® , Provicol® , Calxyl® and Honigum) were tested on equine periodontal fibroblasts. Possible cytotoxic effects were assessed microscopically and by MTT assay, and the expression of inflammatory marker genes was measured by qRT-PCR. RESULTS: PeriCare® and Provicol® had no effects on the cells, whereas Honigum and Calxyl® were associated with severe cytotoxic effects. MAIN LIMITATIONS: The results of this in vitro study need to be confirmed by clinical studies. CONCLUSIONS: Before adapting dental materials from human dentistry, it is crucial to initially test them in a specific equine model.

Within-herd prevalence thresholds for herd-level detection of mastitis pathogens using multiplex real-time PCR in bulk tank milk samples.

The objective of the study was to assess the value of quantitative multiplex real-time PCR examination of bulk tank milk samples for bovine mastitis pathogens as a tool for herd level diagnosis. Using a logistic regression model, this study is aimed at calculating the threshold level of the apparent within-herd prevalence as determined by quarter milk sample cultivation of all lactating cows, thus allowing the detection of a herd positive for a specific pathogen within certain probability levels. A total of 6,335 quarter milk samples were collected and cultured from 1,615 cows on 51 farms in Germany. Bulk tank milk samples were taken from each farm and tested by bacterial culture as well as the commercial PCR assay Mastit 4A (DNA Diagnostic A/S, Risskov, Denmark) identifying Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus agalactiae, and Streptococcus uberis. In addition, PCR was performed on pooled herd milk samples containing milk aliquots from all lactating cows in each of the 51 herds. Only 1 out of the 51 herds was found PCR positive for Streptococcus agalactiae in bulk tank and pooled herd milk samples, and cultured quarter milk samples. Spearman's rank correlations between the cycle threshold value of bulk tank milk PCR and the apparent within-herd prevalence were calculated in regard to Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis. For these pathogens, significant correlations were found. If 1 bulk tank milk sample per herd was tested, the estimated within-herd prevalence thresholds for 90% probability of detection were 27.6% for Staphylococcus aureus, 9.2% for Streptococcus dysgalactiae, and 13.8% for Streptococcus uberis on the cow level. On the quarter level, the within-herd prevalence had to be at least 32.6% for Staphylococcus aureus, 1.7% for Streptococcus dysgalactiae, and 4.3% for Streptococcus uberis to detect a herd as positive using a single bulk milk sample. The results indicate that mastitis pathogens in bulk tank milk can be identified by the applied PCR assay. Bulk tank milk examination is not a reliable tool for the identification of the named pathogens by single testing, but might be a valuable monitoring tool when used frequently with repeated testing. Furthermore, this approach could be a useful monitoring tool for detecting new pathogen occurrence in the herd.

The Relationship between Brachycephalic Head Features in Modern Persian Cats and Dysmorphologies of the Skull and Internal Hydrocephalus.

BACKGROUND: Cat breeders observed a frequent occurrence of internal hydrocephalus in Persian cats with extreme brachycephalic head morphology. OBJECTIVE: To investigate a possible relationship among the grade of brachycephaly, ventricular dilatation, and skull dysmorphologies in Persian cats. ANIMALS: 92 Persian-, 10 Domestic shorthair cats. METHODS: The grade of brachycephaly was determined on skull models based on CT datasets. Cranial measurements were examined with regard to a possible correlation with relative ventricular volume, and cranial capacity. Persians with high (peke-face Persians) and lower grades of brachycephaly (doll-face Persians) were investigated for the presence of skull dysmorphologies. RESULTS: The mean cranial index of the peke-face Persians (0.97 ± 0.14) was significantly higher than the mean cranial index of doll-face Persians (0.66 ± 0.04; P < 0.001). Peke-face Persians had a lower relative nasal bone length (0.15 ± 0.04) compared to doll-face (0.29 ± 0.08; P < 0.001). The endocranial volume was significantly lower in doll-face than peke-face Persians (89.6 ± 1.27% versus 91.76 ± 2.07%; P < 0.001). The cranial index was significantly correlated with this variable (Spearman's r: 0.7; P < 0.0001). Mean ventricle: Brain ratio of the peke-face group (0.159 ± 0.14) was significantly higher compared to doll-face Persians (0.015 ± 0.01; P < 0.001). CONCLUSION AND CLINICAL RELEVANCE: High grades of brachycephaly are also associated with malformations of the calvarial and facial bones as well as dental malformations. As these dysmorphologies can affect animal welfare, the selection for extreme forms of brachycephaly in Persian cats should be reconsidered.

Effect of dietary vitamin E and selenium supplementation on semen quality in Cairn Terriers with normospermia.

Among others, selenium (Se) and vitamin E (VitE) have been provided to dogs to improve semen quality. However, scientific evidence documenting an effect in dogs is lacking. The aim of this study was to investigate the effect of supplementation of these antioxidants on various ejaculate parameters in a randomized, double-blinded trial using Cairn Terrier males exhibiting normal seminal quality parameters. Three dogs each were fed a standardized diet and supplemented with 0.1 mg Se, 100 mg VitE or 0.1 mg Se + 100 mg VitE/dog for 3 months. Ejaculate analyses (volume, progressive motility, vitality, morphology, concentration) were performed before inclusion (D0) and after 1, 2 and 3 months (+1, +2, +3). At the same time, glutathione peroxidase (GSH-PX) and VitE in seminal plasma (SP) and GSH-PX in blood samples were determined. Vitamin E levels in SP were below the detection limit (1.0 mg/L) in all samples. GSH-PX in blood (164.0-2794.4 IU/L) and SP (18.4-4326.0 IU/L) was highly variable. Supplementation only significantly affected the total percentage of sperm head abnormalities (p = .011). Time significantly affected the percentage of morphologically abnormal sperm (p = .025), sperm head abnormalities (p = .007), proximal droplets (p = .001) and GSH-PX in SP (p = .015). Additionally, a significant interaction between time and group was identified for the percentage of membrane-intact sperm (p = .048), head abnormalities (p = .018), acrosomal defects (p = .043) and proximal droplets (p = .002). Although some effects could be identified for selected parameters, we failed to identify a clear trend about how a 3 months VitE and/or Se supplementation affects semen parameters in normospermic Cairn Terriers.

Suspected aspirin resistance in individual healthy adult warmblood horses.

The reasons for this prospective experimental study were to determine a dosing scheme with loading and maintenance dose of aspirin inducing inhibition of platelet function measured by whole blood impedance aggregometry. Ten horses received aspirin orally in the morning with one loading dose of 4.7-5 mg/kg and maintenance doses of 1-1.3 mg/kg daily the following 4 days. Aggregometries (COLtest, ASPItest, ADPtest) and serum salicylic acid were measured. ASPItest showed significant difference in inhibition at 24 and 48 hr (p < .05) and 96 hr (p < .01). Significant change for ADPtest and COLtest couldn't be detected. Serum salicylic acid concentrations were significantly (p < .01) increased at 6 and 12 hr. Despite this, three horses failed any inhibitory effect of platelet function, suspecting an aspirin resistance. Regarding the other seven horses platelet aggregation induced by ASPItest was reduced between 37% and 100% from baseline at 6 and 12 hr and between 0 and 98% during the next 4 days. Correlations of serum concentration of salicylic acid and aggregometries couldn't be detected. It can be presumed that equine platelets are less susceptible to aspirin what may compromise eventually the anticoagulatory effects and efficacy in preventing and treating diseases with increased platelet activation as endotoxaemia or laminitis.

Molecular analyses on Neospora caninum-triggered NETosis in the caprine system.

Neospora caninum is an obligate intracellular protozoan parasite causing serious reproductive disorders in large and small ruminants worldwide. Polymorphonuclear neutrophils (PMN) react against multiple invading pathogens through different mechanisms including the release of neutrophil extracellular traps (NETs). Here, in vitro interactions of caprine PMN and N. caninum tachyzoites were studied. Scanning electron microscopic- and immunofluorescence-analyses demonstrated that caprine PMN undergo NETosis upon contact with tachyzoites of N. caninum, extruding filaments that entrap parasites. Detailed co-localization studies of N. caninum tachyzoite-induced NETs revealed the presence of PMN-derived DNA being decorated with histones (H1, H2A/H2B, H3,H4) and neutrophil elastase (NE) corroborating the molecular characteristics of classical mammalian NETs. As a new result for parasite-induced NETosis, we identified pentraxin and cathepsin B in N. caninum-triggered NETs. Nonetheless, functional inhibition assays revealed that during caprine NET formation triggered by N. caninum different molecular signaling pathways are induced, when compared to other apicomplexan parasites or host species. As such, N. caninum-induced NETosis appears to be influenced by MPO but independent of NADPH oxidase, SOCE, ERK1/2 and p38 MAPK activities. Furthermore, the inhibition of PMN autophagy via blockage of the PI3K-mediated signaling pathway failed to influence tachyzoite-induced NETosis. Since N. caninum-tachyzoites induced caprine NETosis, this effector mechanism should be considered as an early host immune response during acute caprine neosporosis.

Functional anatomy of the equine temporomandibular joint: Collagen fiber texture of the articular surfaces.

In the last decade, the equine masticatory apparatus has received much attention. Numerous studies have emphasized the importance of the temporomandibular joint (TMJ) in the functional process of mastication. However, ultrastructural and histological data providing a basis for biomechanical and histopathological considerations are not available. The aim of the present study was to analyze the architecture of the collagen fiber apparatus in the articular surfaces of the equine TMJ to reveal typical morphological features indicating biomechanical adaptions. Therefore, the collagen fiber alignment was visualized using the split-line technique in 16 adult warmblood horses without any history of TMJ disorders. Within the central two-thirds of the articular surfaces of the articular tubercle, the articular disc and the mandibular head, split-lines ran in a correspondent rostrocaudal direction. In the lateral and medial aspects of these articular surfaces, the split-line pattern varied, displaying curved arrangements in the articular disc and punctual split-lines in the bony components. Mediolateral orientated split-lines were found in the rostral and caudal border of the articular disc and in the mandibular fossa. The complex movements during the equine chewing cycle are likely assigned to different areas of the TMJ. The split-line pattern of the equine TMJ is indicative of a relative movement of the joint components in a preferential rostrocaudal direction which is consigned to the central aspects of the TMJ. The lateral and medial aspects of the articular surfaces provide split-line patterns that indicate movements particularly around a dorsoventral axis.

The effect of endoscopic sterilization on reproductive behavior and pair bond maintenance of feral pigeons (Columba livia).

OBJECTIVE: Problems related to feral pigeons (Columba livia) in cities mainly result from their large numbers due to uncontrolled population growth. The aim of this study was to evaluate whether endoscopic guided sterilization affects the reproductive behavior of feral pigeons under experimental conditions, with the intention of assessing this technique as a potential method for feral pigeon population control. MATERIALS AND METHODS: Five groups of four pairs of feral pigeons each were studied from 8 weeks before, to 7 weeks after sterilization. Both the male and female of the first pair of each group were sterilized, in the second pair only the female and in the third pair only the male was sterilized. The fourth pair acted as a control. All eggs laid were candled to assess fertility. RESULTS: Surgical sterilization had minimal effects on behavior and therefore seems not to have impact on possible field application for population control. All pairs maintained their pair bonds and continued to defend their nesting sites against other pigeons. Only one female copulated with a foreign fertile male while her primary partner was debilitated due to surgery, but returned to him as soon as he recovered. All eggs laid more than 5 days after male sterilization were infertile, whereas all control pairs had fertile eggs. Only one fertile clutch was produced, 5 days after the male's sterilization. Therefore it is assumed that males remain fertile for a limited period of time. CONCLUSION: Endoscopic sterilization seems to be a promising method for field control of feral pigeon populations and sterilization of the male only seems sufficient.

Metabolic signatures of Besnoitia besnoiti-infected endothelial host cells and blockage of key metabolic pathways indicate high glycolytic and glutaminolytic needs of the parasite.

Besnoitia besnoiti is an obligate intracellular and emerging coccidian parasite of cattle with a significant economic impact on cattle industry. During acute infection, fast-proliferating tachyzoites are continuously formed mainly in endothelial host cells of infected animals. Given that offspring formation is a highly energy and cell building block demanding process, the parasite needs to exploit host cellular metabolism to meet its metabolic demands. Here, we analyzed the metabolic signatures of B. besnoiti-infected endothelial host cells and aimed to influence parasite proliferation by inhibitors of specific metabolic pathways. The following inhibitors were tested: fluoro 2-deoxy-D-glucose and 2-deoxy-D-glucose (FDG, DG; inhibitors of glycolysis), 6-diazo-5-oxo-L-norleucin (DON; inhibitor of glutaminolysis), dichloroacetate (DCA; inhibitor of pyruvate dehydrogenase kinase which favorites channeling of glucose carbons into the TCA cycle) and adenosine-monophosphate (AMP; inhibitor of ribose 5-P synthesis). Overall, B. besnoiti infections of bovine endothelial cells induced a significant and infection rate-dependent increase of glucose, lactate, glutamine, glutamate, pyruvate, alanine, and serine conversion rates which together indicate a parasite-triggered up-regulation of glycolysis and glutaminolysis. Thus, addition of DON, FDG, and DG into the cultivation medium of B. besnoiti infected endothelial cells led to a dose-dependent inhibition of parasite replication (4 µM DON, 99.5 % inhibition; 2 mM FDG, 99.1 % inhibition; 2 mM DG, 93 % inhibition; and 8 mM DCA, 71.9 % inhibition). In contrast, AMP had no significant effects on total tachyzoite production up to a concentration of 20 mM. Together, these data may open new strategies for the development of therapeutics for B. besnoiti infections.