Associations between minerals and metabolic indicators in maternal blood pre- and postpartum with ewe body condition, methane emissions, and lamb body weight development.

In sheep production, economic efficiency strongly depends on the maternal health and feed efficiency status and on weaning performances of their offspring. Accordingly, an optimal level for the supply with macro- and microelements and the ewe energy status has impact on the fetal development during gestation and on maternal milk production during lactation. Furthermore, this study addressed intergenerational aspects, i.e., on associations between maternal energy metabolism profiles considering the macro- and microelement status, metabolic indicators (e.g. ß-hydroxybutyrate (BHB)), body condition and methane (CH4) emissions with lamb BW (LBW) in two sheep breeds. Traits were recorded at the beginning of gestation (ewe traits), at lambing, three weeks postpartum, and at weaning (ewe and lamb traits). Trait recording included CH4 emissions (recorded via laser methane detector (LMD)), ewe BW (EBW), backfat thickness (BFT), and body condition score (BCS) from 46 ewes (24 Merinoland- (ML), 22 Rhönsheep (RH)), and LBW of their 87 (35 ML, 52 RH) purebred lambs. Serum levels of the following ewe blood parameters were determined: calcium (Ca), sodium (Na), potassium (K), phosphate (P), nonesterified fatty acids (NEFA), BHB, glutamate dehydrogenase (GLDH), selenium (Se), copper (Cu), iron (Fe), zinc (Zn), and magnesium (Mg). Mixed models were applied to infer associations between ewe blood parameters with EBW, BFT, BCS, and CH4 and with LBW recorded in offspring. At weaning, a maternal serum Mg level > 1.0 mmol/L was significantly associated with an increase of 13% in LBW in ML, compared to offspring from ML ewes with a serum Mg concentration within the lower reference range (0.8 mmol/L). Furthermore, higher Cu levels were favorably associated with ewe BCS and BFT at weaning in both breeds. In RH ewes, a Se level > 2.4 µmol/l was significantly associated with increased BCS. In the ML breed, high Zn levels during lactation were associated with reduced CH4 emissions. Ewe EBW was significantly larger for ML ewes representing low Ca levels. A low BHB level was associated with decreasing CH4 emissions in RH and ML. Serum levels for Na, K, P, GLDH, and Fe did not significantly affect the traits of interest. Trait associations from the present study indicate the importance of the mineral supply and metabolic status of the ewe with regard to body condition, CH4 emissions, and LBW development, but depending on the breed. Identified associations might contribute to energy efficiency in sheep production systems.

Semen analysis and successful artificial insemination in the St. Vincent amazon (Amazona guildingii).

The St. Vincent amazon (Amazona guildingii) is an endemic parrot on the Carribean island St Vincent. Due to poaching, trade, natural events such as hurricanes and habitat loss the species declined severely throughout the 20th century to a total number of 487 adult individuals and is currently regarded as vulnerable by IUCN. Captive breeding is attempted in terms of species conservation, but reproduction rates have been low due to reproductive problems such as mate aggression, asynchronous reproductive activity and infertile eggs. The aims of the present study were; firstly, to evaluate whether semen analysis might help to assess the fertility of males and to detect potential reasons for infertile eggs; secondly, to increase the number of offspring using artificial insemination, and as a future effect, to increase the presence of genetically valuable males in the ex-situ breeding population. For semen collection electric stimulation was used in 15 mature and healthy St. Vincent amazons with a success rate of 89% (202/227 attempts) in 14/15 males. Quality assessment of the semen included the evaluation of volume, pH, color, consistency and contaminations of the ejaculate, as well as estimation of motility, viability, morphology, concentration and total count of spermatozoa. Semen pH ranged from 6.7 to 7.5. Median sperm motility was 50% and median progressively forward motility 40%. Mean sperm concentration (x¯ ± SD) was 21,313.5 ± 22,408.8 spermatozoa/µl and mean sperm viability 66 ± 16%. Semen samples contained on average 20.5% morphologically normal spermatozoa and sperm malformations were detected mainly in the head (x¯ = 47.6%) and the tail regions (x¯ = 27.7%). Interestingly round bodies were detected in the ejaculates with a mean ratio of 0.6 round bodies per sperm. Semen analysis proved to be very useful to identify males with poor semen quality. Artificial insemination was performed 46 times in 9 females with either individual or pooled semen samples and 13 eggs from 7 females were laid afterwards. In 3 eggs, embryonic development was detected and 1 chick hatched successfully. Paternity testing confirmed the fatherhood of a one-winged semen donor male, a bird which was not able to copulate naturally. The results are very promising and underline that assisted reproduction techniques are a suitable tool for species conservation in captive breeding programs for psittacines.

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.

[Quality Assessment of Bovine and Equine Colostrum - An Overview]. / Qualitätsbeurteilung von bovinen und equinen Kolostrum ­ Eine Übersicht.

INTRODUCTION: Good quality colostrum intake is essential for passive immunity in neonatal calf and foals. The quality of colostrum depends on its IgG content. Since in practice the possibilities for a direct determination of these parameter are limited, an estimation of colostrum quality is often based on its biophysical properties. The simple methodology allows to perform measurements directly on site and immediately after birth. The biophysical properties included in the current review are density, refractive index, relative density, viscosity, pH, and color. The determination of the density of bovine colostrum is a common method for the evaluation of the IgG content in practice since it correlates with IgG. Density of bovine colostrum should be 1047 g/l. Different colostrometers are available for the measurement. This method is also used in the horses. Density of equine colostrum should be 1060 g/l. Refractive index or the relative density by refractometer has been proven to correlate with the IgG and is an established method in cattle and horses. Good bovine colostrum should have an index of 22% brix and equine colostrum 23% brix. Although not yet established in practice an additional methodology would be the determination viscosity of bovine colostrum. Viscosity could be measured with an outlet cup. A relationship between viscosity and IgG has demonstrated. Sufficient data for this method is missing for equine colostrum. Color and pH are not suitable for estimating the IgG content.

INTRODUCTION: La prise de colostrum de bonne qualité est essentielle pour la construction d'une immunité passive chez le veau et le poulain nouveau-nés. La qualité du colostrum dépend de sa teneur en IgG. Comme dans la pratique les possibilités de détermination directe de ce paramètre sont limitées, une estimation de la qualité du colostrum est souvent basée sur ses propriétés biophysiques. Grâce à une méthodologie simple, la mesure peut généralement être effectuée directement sur place et immédiatement après la naissance. Les propriétés biophysiques incluses dans cette revue sont la densité, l'indice de réfraction, la densité relative, la viscosité, le pH et la couleur. La détermination de la densité du colostrum bovin est en pratique une méthode courante pour évaluer la teneur en IgG, car la densité est en corrélation avec la teneur en IgG. La densité du colostrum doit être d'au moins 1047 g/l. Différents colostromètres sont disponibles pour cette mesure. Cette méthode est également utilisée chez le cheval. La densité du colostrum équin doit être d'au moins 1060 g/l. La détermination de l'indice de réfraction ou de la densité relative au moyen d'un réfractomètre est une procédure établie chez les bovins et les chevaux, car une corrélation claire avec la teneur en IgG est prouvée. Pour être considéré comme bon, le colostrum bovin doit atteindre une valeur de 22% et le colostrum équin une valeur de 23% sur l'échelle de Brix. L'évaluation du colostrum bovin par viscosité est une autre approche qui doit encore être mise en pratique. Là encore, une relation entre la viscosité et la teneur en IgG pourrait être prouvée. La mesure de la viscosité peut être effectuée à l'aide d'une coupe d'écoulement. Chez le cheval, on ne dispose pas encore de données suffisantes sur cette méthode. La valeur du pH et la couleur ne conviennent pas pour estimer la teneur en IgG.

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.

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.

Bacteriological findings in the canine uterus during Caesarean section performed due to dystocia and their correlation to puppy mortality at the time of parturition.

Canine intrauterine bacteriological flora during dystocia is unknown. Thus, frequency (bacterial growth (not) detected), quality (species and number of different bacterial isolates) and quantity (colony-forming units) of intrauterine bacteria in relation to in utero foetal death in 50 bitches undergoing emergency Caesarean section were investigated. Bacterial growth was quantified from single colonies, (+) (0.5), to strong growth, +++ (3) and was observed in 34 bitches (68%), with Staph. epidermidis (n = 12), Staph. intermedius-group (n = 7), ß-haemolytic streptococci (n = 6), Staph. aureus, α- and γ-haemolytic streptococci (n = 4 each) being most common and one to four bacteria per sample. Regarding the quantity, most often (n = 46) low growth was identified. In bitches with living pups only (group I), mean number of isolates was 0.78 ± 0.83 compared to 1.60 ± 1.10 (living + stillborn pups, group II) and 1.0 ± 1.15 (stillborn pups only, group III) and mean bacterial growth in groups I/II/III was + (1.0, quantity), + (1.4) and ++ (1.6). Taking just positive samples into consideration, mean number of bacterial isolates was significantly higher in group II compared to I (p = .0088). We concluded that the canine uterus cannot be considered free of bacteria during dystocia. Mean numbers of different bacterial isolates and quantity of bacterial growth are higher in bitches with in utero foetal death.

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.

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.

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.

Seasonal and genera-specific variations in semen availability and semen characteristics in large parrots.

In large parrots electro-stimulation is suitable for collecting semen, and therefore, to facilitate semen examination and artificial insemination. Previous studies have detected differences in the semen collection success rate and semen parameters between psittacine genera. It remained unclear whether these differences were genera-related, seasonal variations or depend on the males' relationship status. To answer these questions, semen collection and spermatological analysis were performed for four psittacine groups (macaws, amazons, eclectus parrots and cockatoos) over 13 months. In one breeding facility, semen collection was attempted in 82 males using electro-stimulation twice monthly. A complete spermatological evaluation was performed on 435 semen samples. Volume, color, consistency, contamination and pH of semen, as well as motility, progressive motility, sperm concentration, total sperm count, viability, and morphology of spermatozoa were evaluated. Seasonality affected the collection success rate in macaws and amazons. Thereby, in amazons a distinct peak was observed several days before and around oviposition, whereas eclectus parrots and cockatoos produced semen all year round. The average sperm concentration was highest in eclectus parrots (2.7 × 106 sperm/µl) and lowest in macaws (35.6 × 103 sperm/µl). The differences in the semen collection success rate and semen parameters seem to coincide with the bird's breeding biology. The collected data allows a prognostic estimation when semen collection seems favorable, and may be taken as orientation values for semen analysis in these species.

Parturition induction in ewes by a progesterone receptor blocker, aglepristone, and subsequent neonatal survival: Preliminary results.

The clinical effects of aglepristone treatment to induce parturition in ewes and their newborns were reported. Three experimental groups were defined: group AG5 (n = 5), group AG10 (n = 5), and group CG (n = 5) in which ewes were injected twice with 5, 10 mg/kg of aglepristone, and saline solution of ewes, respectively. Different parameters associated with parturition in ewes and their newborns were investigated. Serum progesterone, oxytocin, and free and conjugated total estrogens were measured after treatments until parturition. No statistical difference was found from first aglepristone administration to onset of lambing between AG5 and AG10 (23.90 ± 6.20, 40.00 ± 6.71 hours). Parturition induction in two groups shortened the gestational length significantly compared with the control group (P = 0.003). Dystocia was observed in two ewes in group AG10. The placental weight showed statistically significant difference only between the AG10 and CG (P = 0.039), but no difference was observed in the placental expulsion period between the groups. Decrease in food consumption 24 to 36 hours after parturition in all ewes and skin necrosis in an ewe in group AG5 were observed. Progesterone concentration was significantly lower in AG5 than that in ewes in group AG10 and CG (P < 0.05). No difference was observed in concentrations of free total estrogens and oxytocin between groups. The body temperature of lambs was significantly different between AG10 and CG groups both right after (P = 0.011) and 12 hours after parturition (P = 0.014). The lambs in CG had the highest mean birth weight (4.29 ± 0.28 kg), which was significantly different from the induced groups. No significant difference of blood pH and blood gases values between groups was identified both at birth and 12 hours after parturition for lambs. Significant differences could clearly be observed in total protein and blood urea nitrogen and total protein findings 12 hours after parturition (P < 0.05), whereas no difference was found in blood glucose, albumin, inorganic phosphor, triglyceride, or total cholesterol parameters. The results of this study show that the administration of aglepristone to induce parturition can precisely control lambing time without any side effects in either mothers or lambs.

Effect of seminal plasma vesicular structures in canine frozen-thawed semen.

Membrane vesicles (MVs) in the ejaculate have been identified in various species and are considered to affect membrane fluidity due to their characteristic molecular composition. Addition of MV to human frozen semen has been shown to improve post-thaw motility. Similarly, a beneficial effect has been suggested for frozen equine semen. As post-thaw canine semen quality varies widely between dogs, the aim of our study was to test for the effect of addition of canine MV on post-thaw semen quality in dogs. Semen samples from 10 male dogs were purified from MV and prepared for freezing. In experiment 1, three groups were compared: sperm frozen (1) with MV (S1); (2) without MV, but MV added immediately after thawing (S2); and (3) without MV (C). Semen analysis included computer-assisted sperm analysis of motility parameters immediately after thawing (t0), after 10 (t10) and 30 minutes (t30), % living sperm, % membrane intact, % morphologically normal sperm (all t0 and t30). Computer-assisted sperm analysis motility distance and velocity parameters (all P < 0.05) and % living sperm (P < 0.001) were significantly affected by treatment with a temporary increase of distance and velocity parameters at t0 to t10, but a significant decrease of the aforementioned parameters at t30 in samples with MV. In experiment 2, different MV protein concentrations added after thawing were compared: 0.05 mg, 0.1 mg, and 0.2 mg/mL. Computer-assisted sperm motility analysis was performed at t0, t10, and t30. No differences between MV concentrations were identified, only a significant interaction between effect of treatment and time for progressive motility (P < 0.01). Our study identified a short-term beneficial effect of canine MV on post-thaw distance and velocity parameters, whereas at t30 progressive motility, motility parameters and % living sperm were reduced in samples with MV compared to C. The results point to species-specific differences regarding the MV effect on frozen semen and indicate the need for further studies using different semen and MV purification protocols and more frequent analyses. At the moment, addition of MV is not an option to improve post-thaw semen quality in dogs.

Morphological characterisation of vesicular structures in the canine ejaculate.

Membrane vesicles (MV) have been identified in seminal plasma from various species and they are thought to have a significant impact on semen quality and fertilisation. Although recently presence of MV has been also described in the canine ejaculate, detailed knowledge on their morphology is missing by now. This is, however, needed to provide a basis for detailed biochemical and functional studies as it is generally assumed that different MV populations are responsible for distinct tasks. MV were prepared for light (LM) and transmission electron microscopy (TEM) analysis using samples from normospermic dogs (n=15), hypokinozoospermic dogs (n=2, h) and one castrated azoospermic dog (a). For TEM, a new preparation protocol was used resulting in a higher MV retrieval rate. Using fractionated semen samples, most MV were identified in the second (sperm-rich) fraction in LM. Using pooled ejaculates, three different MV types could be identified in LM: (1) large MV with a marginal accumulation of opaque, granulated material, (2) medium- to small size MV with dense, opaque content and (3) small MV with no further defined contents. No direct contact between sperm and MV could be visualised. In TEM, 11 different MV types were identified based on diameter, structure, contents and electron density of contents as well as presence, number and size of smaller MV inside the MV itself. In normospermic males, secondary vesicles (type i, H, K1/2) included smaller vesicles and had a weighted mean diameter of 409.46 nm; hereof types i, H and K1 were smaller (mean: 287.55 nm, range: 51.25-994.86 nm) and type K2 was larger (mean: 1746.43 nm, range: 1003.66-3289.34 nm). Primary vesicles (mean diameter: 135.29 nm) - without vesicles inside - were differentiated into larger MV (A, B, C1/2) with a mean diameter of 219.63 nm (range: 39.08-1300.13 nm) and small primary MV (F, G) with a mean diameter of 66.12 nm (range: 24.62-99.84 nm). Whereas all mentioned MV were round to oval and mostly double-, rarely multiple-membrane surrounded, one longish primary MV type (L) was identified. In general, small primary vesicles were most common independent of semen quality, but distribution frequency of vesicle types differed between normospermic, pathospermic dogs and the castrated male. Mean weighted diameter of MV was 195.14 nm (range: 24.62-3289.34 nm) in normospermic males with the maximum diameter being smaller in the other dogs (h: 2096.78 nm; a: 1314.06 nm). Our results provide new information about ultrastructure and distribution frequency of canine MV in normospermic males and point to possible differences in MVs depending on semen quality. They provide the basis for further detailed functional analysis of MV subpopulations. Furthermore, the presence of MV in the castrated azoospermic male confirms an at least partly prostatic origin of canine MV.

[Death of two mares with retained fetal membranes]. / Tod zweier Stuten mit Retentio secundinarum.

Retained fetal membranes in mares is a relative frequent disease which can lead to severe complications. Two case studies are described in which mares died acutely despite intensive care. Pathological examination confirmed severe hemorrhage as the cause of death in both animals.

[Risk factors of increased abortions in dairy herds - a statistical analysis]. / Risikofaktoren für ein erhöhtes Abortvorkommen in Milchviehbetrieben - eine statistische Analyse.

OBJECTIVE: To examine whether dairy herds with and without abortion problems differ in their management and prevalence of antibodies against various pathogens, through collecting and analysing data from these herds. MATERIAL AND METHODS: Data on management parameters, clinical examinations and serological results were obtained from 48 dairy herds in middle Hessen. Information on the extent of stock renewal, form of animal housing, type of herd occupancy and use of a maternity pen was compiled. A random sample was taken from every herd, consisting of a minimum of three healthy non-lactating cows, three clinically sound puerperal animals, three animals having had an abortion and three clinically healthy pregnant heifers. In these animals, the body condition score was evaluated and a clinical gynaecological examination was performed. Additionally, these animals were serologically examined for the presence of antibodies against the bovine viral diarrhoea virus, Coxiella, Listeria and Neosporacaninum. A comparison between the herds with and without an abortion problem was performed. RESULTS: Data of 591 animals from 48 farms was compiled. Thirty-eight herds had an abortion problem. In the affected farms, a maternity pen was used significantly more often and cattle were significantly more frequently housed in free stalls when compared to farms without an abortion problem (p < 0.05). The number of animals serologically positive for Coxiella was significantly higher in herds with abortion problems (p = 0.03). No significant difference was found between the occurrence of antibodies against Listeria and Neospora caninum in herds with or without abortion problems, respectively (p > 0.05). CONCLUSIONS: It was identified that in addition to infectious agents, management/husbandry factors that promote the spread of disease are a risk factor for abortion in dairy cattle. Besides taking steps against pathogens, farms with abortion problems have to implement measures to minimize the spread of infection between animals. A critical control point is likely to be the maternity pen.

[Incidence of puerperal diseases during the first 10 days after foaling in the mare]. / Häufigkeit puerperaler Erkrankungen während der ersten 10 Tage post partum bei Stuten.

OBJECTIVE: The aim of this study was to evaluate the frequency of puerperal diseases in breeding mares in the first 10 days after birth by analysing patient data. MATERIAL AND METHODS: In a university clinic patient data of 308 breeding mares with puerperal disorders which presented within the first 10 days postpartum were evaluated over a period of 10 years. A distinction was made between diseases which were able to be diagnosed at the first examination and diseases which developed during the patient's stay in the clinic. RESULTS: A total of 21 diseases were diagnosed, with a retained placenta, lochiometra and injuries to the perineum being the most common. Many mares displayed more than one disease. Mares with a retained placenta most commonly also presented with perineal ruptures, followed by animals who also had lochiometra. Mares suffering from lochiometra commonly presented together with a retained placenta and injuries as a result of birth. Some of the mares developed further diseases. In mares with a retained placenta, this was most commonly lochiometra, followed by puerperal laminitis and thrombophlebitis. CONCLUSION AND CLINICAL RELEVANCE: The data collection shows that several diseases could relatively frequently be diagnosed in mares with puerperal disorders. Therefore, a higher percentage of further diseases must be assumed for mares which have a puerperal disease.

[Perineal hypospadia in a red Holstein-calf. A case report]. / Perineale Hypospadie bei einem rotbunten Deutsche-Holstein-Kalb. Ein Fallbericht.

Hypospadia is a congenital deformation of the urethra. Several surgical correction methods have been described, but there are also cases when an operation is not necessarily required. In this report, the case of a bull calf which did not need an invasive therapy will be presented.

Comparison of accuracy of pregnancy-associated glycoprotein (PAG) concentration in blood and milk for early pregnancy diagnosis in cows.

The aim of this study was to compare two methods of early pregnancy diagnosis by determining pregnancy-associated glycoprotein (PAG) concentration in blood and PAG concentration in milk. Blood and milk samples were obtained on days 0 (AI day), 14, 21, 28, 35, 49, 63, 77, 91 and 105 of gestation from 60 lactating Holstein Frisian cows from one herd, carrying live fetuses. To determine PAG concentration a specific RIA system (RIA-706) was used. PAG concentration in blood and milk increased after 28 days of pregnancy, with blood concentrations being significantly higher than in milk. However, the accuracy of both tests at this time point was similar (sensitivity: 92 % in blood, 93 % in milk; specificity 53 % and 60 % respectively). None of the tests were able to detect open cows properly at this stage. On day 35 of gestation sensitivity (100 % for blood, 97 % for milk) and specificity (100 % for blood, 100 % for milk) were high enough to be used for reliable pregnancy diagnosis. The accuracy (sensitivity and specificity) for PAG concentrations in blood and milk for the rest of the study was 100 %. Our investigation shows that PAG determination in milk is a stress-free and non-invasive method for early pregnancy diagnosis in cattle.

Le but de cette étude était de comparer les concentrations de glycoprotéines associées à la gestation (PAG) dans le sang et le lait en vue d'un diagnostic de gestation précoce chez la vache. Des échantillons de sang et de lait ont été prélevés aux jours 0 (IA), 14, 21, 28, 35, 49, 63, 77, 91 et 105 de la gestation sur 60 vaches Holstein Frisonnes en lactation avec un fœtus vivant. On a utilisé pour mesurer la concentration de PAG une méthode RIA spécifique (RIA-706). Les concentrations de PAG dans le sang et le lait s'élevaient à partir du 28ème jour de gestation, les concentrations mesurées dans le sang étant nettement plus élevée que celles mesurées dans le lait. L'exactitude des deux tests était à ce moment-là similaires (sensitivité: 92 % dans le sang, 93 % dans le lait, spécificité: 53 % respectivement 60 %). Aucune des deux méthodes n'était, à ce stade, à même de distinguer une vache non portante avec justesse. Au 35ème jour de gestation, la sensitivité des deux méthodes (100 % dans le sang, 97 % dans le lait) et leur spécificité (100 % dans le sang, 100 % dans le lait) était assez élevées pour permettre un diagnostic de gestation sûr. L'exactitude (sensitivité et spécificité) des mesures de concentration de PAG dans le sang et le lait durant la suite de la gestation était de 100 %. Ces études montrent donc que la mesure de PAG dans le lait représente une méthode non-invasive et exempte de stress pour le diagnostic précoce de gestation chez la vache.

[Uterine torsion in cattle--therapy and consequences for calf and cow]. / Torsio uteri beim Rind--Therapie und Folgen für das Kalb und die Kuh.

AIM: To summarize the available literature on the therapy of uterine torsion in cattle and the consequences for cow and calf. MATERIAL AND METHODS: Analysis of the literature using electronic libraries (PubMed, Medline), German veterinary medical journals and obstetrical textbooks. RESULTS: The therapy includes the attempt to rotate the uterus back into its physiological position. Direct and indirect methods of retorsion are available and applied according to the case conditions. Subsequently, the extraction of the calf can be performed via vaginal delivery or caesarean section. The presence of uterine torsion always leads to dystocia. Following a successful retorsion, the time and degree of uterine torsion strongly influence the progress of the birth. The prognosis also depends on the aforementioned factors and varies between good to unsuccessful. The vitality of the calf displays great variation depending on the literature (14-90%), however, is generally greater under field than clinical conditions. Focussing on the puerperal development of the cow, all grades from mild irritations of the uterine involution to fatal complications occur. The influence on fertility depends on the progress of the birth and existing secondary complications. The risk for electrolyte disturbances is increased (approximately 50%) as is the risk of birth-associated injuries (approximately 20%). The incidence of placental retention varies widely between different authors (3-52%).