The metabolite products of chlorocholine chloride (CCC) in eggs and meat of laying hens fed 15N-CCC containing diets

An experiment was conducted to evaluate the metabolic products of chlorocholine chloride (CCC) in eggs and meat of laying hens fed a diet containing (15)N-CCC. Ten brown laying hens were randomly divided into two groups of five each. One group was offered (15)N-CCC free diet while the other group received a diet with 100 ppm (15)N-CCC for 11 days. Samples of eggs and meat from the laying hens were collected. Egg yolks and albumen were separated. Meat was collected from the breast and femur. The metabolic products of CCC were measured using ion trap electrospray ionisation mass spectrometry (ion trap-ESI-MS/MS). Determination of CCC or its metabolites in eggs and meat showed that CCC was metabolised to choline. Corresponding MS/MS spectra were obtained for m/z 104 (choline) or 105 ((15)N-choline), whereas nothing was detected at m/z 122 (CCC) or 123 ((15)N-CCC). The results from this study indicate that CCC will be metabolised in tissues of laying hens.     

Echinococcus multilocularis in Austrian foxes from 1991 until 2004

The prevalence rates of Echinococcus multilocularis in foxes (n=5600) evaluated in several Austrian surveys conducted between 1991 and 2004 were analysed for spatial and temporal differences. Data from early studies (1993-1997) in which the intestinal scraping technique (IST) was utilized were compared with data from recent (1999-2004) investigations, which made use of the shaking in a vessel technique (SVT), and it was assessed whether or not the infection rates of Austrian foxes had increased between the investigated intervals. In total, data from 85 districts are presented and both the retrospective and recent data are available from 39 of these districts. A Bayesian hierarchical model of parasite prevalences is presented which (i) accounts for differences in the sensitivity of IST and SVT, (ii) incorporates spatial auto-correlation between neighbouring districts, (iii) investigates the possibility of a temporal shift in the infection status of foxes, and (iv) quantifies uncertainty at each level of the model. The national average prevalence rates in the mid-1990s and at the turn of the millennium were 2.4% (95% confidence intervals 1.1-4.8) and 3.9% (95% confidence intervals 1.5-8.4) respectively. Above average prevalence rates were observed in the western and the northern parts of the country. Evidence is also presented for a temporal augmentation of the prevalence rates in some districts in the northern and eastern parts of the country. These findings are in concordance with several investigations in other European states where both newly emerged areas and elevated levels of transmission in existing endemic areas have been found. None of the districts investigated here showed significant evidence of a drop in prevalence.     

Detection of chlamydiae in boar semen and genital tracts

Chlamydiae cause abortion and reproductive disorders in sows. Although organisms can infect the male genital tract, little is known about the disease situation in boars. Hence, we examined the prevalence of chlamydial infection in semen and genital tracts of boars. Samples collected from Swiss boars (group A: n=42), and boars from Germany (group B: n=39) were examined by bacteriology, LPS-ELISA, immunohistochemistry (IHC) and polymerase chain reaction (PCR). The latter methodology involved use of three PCR assays including 16Sig rDNA, IGS-S (intergenic spacer 16S/23S-Short) and IGS-L (intergenic spacer 16S/23S-Long) PCR for comparison methods. PCR sensitivity and the presence of potential PCR inhibitors were determined by spiking semen with Chlamydophila (Cp.) abortus DNA. Detection limits of the 16Sig and IGS-S PCR were 10 templates, while the IGS-L PCR was less sensitive (100 templates). Of 25 semen samples that were collected from group A, one semen sample was positive for Cp. psittaci and two were positive for Chlamydia-like organisms by 16Sig PCR. Screening of sera from Swiss boars revealed three animals with positive reactions in the LPS-ELISA, although we failed to detect chlamydiae within organs of these or sera-negative animals by IHC or IGS-S PCR. In group B, 10 ejaculates were positive for Chlamydia (C.) suis and two were positive for Chlamydia-like organisms by 16S PCR. The identification of DNA from Chlamydia-like organisms in semen from both groups of boars was surprising and a role for these bacteria in reproductive diseases requires further assessment. In conclusion, the prevalence of chlamydial infection was low in group A animals indicating that venereal transmission may not be significant for Chlamydia-associated reproductive diseases in pigs, although rare cases may occur.     

Survival of mouse embryos after vitrification depending on the cooling rate of the cryoprotectant solution

The aim of the study was to determine the relationship between the rate of cooling of eight-cell mouse embryos to the temperature of liquid nitrogen (-196 degrees C) and their developmental capacity after thawing on the basis of their ability to leave the zona pellucida ('hatching') during in vitro culturing. Eight-cell embryos were obtained from superovulated female mice and divided into three experimental and one control group. Embryos from the experimental groups were cryopreserved by the vitrification method using ethylene glycol as cryoprotectant. The vitrification protocols used in the study differed in the rate of cooling of the cryoprotectant solution. Embryos from the first group were frozen in conventional 0.25-ml plastic straws, those from the second group in pipetting 'tips', and embryos from the third group, placed in vitrification solution, were introduced dropwise directly into liquid nitrogen. The control group of embryos was cultured in vitro without freezing in a culturing medium in an environment consisting of 95% air and 5% CO2. The developmental capacity of thawed embryos was assessed on the basis of their ability to leave the zona pellucida ('hatching') after three days of in vitro culturing. In the control group 95.1% of embryos 'hatched'. A significantly higher number of embryos that 'hatched' after thawing was observed in the group introduced dropwise directly into liquid nitrogen (60.0%) compared to the group frozen in pipetting 'tips' (37.9%). The group frozen in straws yielded significantly the lowest proportion of 'hatching' embryos (8.1%). These results showed that increasing cooling rates during vitrification of embryos improved their survival.     

Partial prostatectomy using Nd:YAG laser for management of canine prostate carcinoma

OBJECTIVE: To report a technique for partial prostatectomy by laser dissection and to evaluate outcome and complications in dogs with prostate carcinoma (PCA). STUDY DESIGN: Experimental and clinical case series. ANIMALS: Four normal dogs and 8 dogs with PCA. METHODS: Subcapsular partial prostatectomy, sparing the urethra and the dorsal aspect of the prostatic capsule, using Nd:YAG laser dissection to remove the prostatic parenchyma and control hemorrhage was performed in 4 normal dogs and subsequently in 8 dogs with histologically confirmed PCA. Additional treatment of PCA dogs included local application of interleukin-2 and systemic administration of meloxicam. Prostate size, complications, and survival time were recorded. Laser-associated thermal damage to surrounding tissue was evaluated by histology. RESULTS: In normal dogs, no damage to the dorsal prostatic capsule or urethra was detected. In PCA dogs, median survival was 103 days (range, 5-239 days). Three dogs died from complications within 16 days, whereas 5 (median survival, 183 days; range, 91-239 days) had improvement or resolution of clinical signs. Urinary incontinence did not occur. CONCLUSION: Laser assisted subcapsular partial prostatectomy can be performed in dogs with PCA without development of postoperative incontinence. CLINICAL RELEVANCE: Subcapsular partial prostatectomy is a potential palliative treatment for PCA in dogs and may lead to the resolution of clinical signs for several months.     

Case report: congenital myoclonus in a German Holstein calf

A female German Holstein calf was not able to stand up after birth. Resting the animal was lying in normal position and could lift its head. Sensory stimuli like auditory or tactile impulses induced myoclonic jerking of the whole body. Afterwards it calmed down quickly. The signs observed correspond to the clinical findings of congenital myoclonus in poll Hereford calves. The pathological examination revealed no indications for changes in organs. The inbreeding coefficient of the calf was 1.56 %. The present type of congenital myoclonus in the calf examined is likely to be genetically determined, even if the point mutation in exon 2 of the glycin receptor alpha 1 gene was not confirmed.     

Regulation by Jun N-terminal kinase/stress activated protein kinase of cytokine expression in Mycobacterium avium subsp paratuberculosis-infected bovine monocytes

OBJECTIVE: To evaluate activation of Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) pathway in bovine monocytes after incubation with Mycobacterium avium subsp paratuberculosis (Mptb) organisms. SAMPLE POPULATION: Bovine monocytes obtained from 4 healthy adult Holstein dairy cows. PROCEDURES: Bovine monocytes were incubated with Mptb organisms with or without a specific inhibitor of the JNK/SAPK pathway (SP600125) for 2, 6, 24, or 72 hours. Expression of interleukin (IL)-1beta, IL-10, IL-12, IL-18; transforming growth factor-beta (TGF-beta); and tumor necrosis factor-alpha (TNF-alpha) and the capacity of Mptb-infected monocytes to acidify phagosomes and kill Mptb organisms were evaluated. Phosphorylation status of JNK/SAPK was evaluated at 10, 30, and 60 minutes after Mptb incubation. RESULTS: Compared with uninfected control monocytes, Mptb-infected monocytes had increased expression of IL-10 at 2 and 6 hours after incubation and had increased expression of TNF-alpha, IL-1beta, IL-18, and TGF-beta at 2, 4, and 6 hours. Additionally, Mptb-infected monocytes had increased expression of IL-12 at 6 and 24 hours. Addition of SP600125 (specific chemical inhibitor of JNK/SAPK) resulted in a decrease in TNF-alpha expression at 2, 6, and 24 hours, compared with untreated Mptb-infected cells. Addition of SP600125 resulted in a decrease in TGF-beta expression at 24 hours and an increase in IL-18 expression at 6 hours. Addition of SP600125 failed to alter phagosome acidification but did enhance the capacity of monocytes to kill Mptb organisms. CONCLUSIONS AND CLINICAL RELEVANCE: Activation of JNK/SAPK may be an important mechanism used by Mptb to regulate cytokine expression in bovine monocytes for survival and to alter inflammatory and immune responses.     

Sex steroids level in blood plasma and ovarian follicles of the chimeric chicken

The study was performed to determine the hormonal status of mature germline chimeras obtained by blastodermal cell transfer from chicken embryos of a donor breed [Green-legged Partridgelike breed (GP) x Araucana (AR)] to those of a recipient breed [White Leghorn (WL)] being at the same stage of embryonic development. The egg-laying chimeras and WL hens (control) of the same age were used in the experiment. At first, blood samples were taken from each bird at 0.5, 5, 12.5 and 18.5 h following oviposition. Subsequently, the chimeras and the WL hens were decapitated 1-2 h after ovulation. A stroma and the following follicles were isolated from the ovary: white normal (1-4, 4-6 and 6-8 mm), white atretic and yellow preovulatory follicles (F4-F1). Sex hormones, progesterone (P4), testosterone (T) and oestradiol (E2) in blood plasma and ovarian follicles were determined radioimmunologically. The activity of the 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in the granulosa and theca layers of the follicles was analysed histochemically. In chimeric chickens, a higher level of T in blood plasma during the ovulatory cycle was noticed. However, in the stroma, white prehierarchical and medium-size preovulatory ovarian follicles the level of T was significantly lower. With respect to E2, its elevated levels were found both in blood and in the ovarian follicles. There were no significant differences in P4 concentrations in blood plasma while in ovarian follicles a higher level was observed only in white 6-8 mm follicles. 3beta-HSD activity in granulosa and theca layers of the ovarian follicles in chimeras was not different from that in the WL hens. In conclusion, the results obtained indicate that germline chimeras exhibit significant alterations in sex hormone levels in the ovary and blood plasma, which in turn may affect their reproductive abilities.     

Evaluation of ilial screw loosening after triple pelvic osteotomy in dogs: 227 cases (1991-1999)

OBJECTIVE: To investigate factors influencing screw loosening after triple pelvic osteotomy (TPO) and ischial wire stabilization of the acetabular segment. DESIGN: Retrospective case series. Animals-227 dogs with congenital hip dysplasia or subluxated hip joints. PROCEDURES: Medical records and radiographs of 227 dogs that underwent 332 TPO procedures were evaluated, and data pertaining to screw type, plate position, sacral screw engagement, use of ischial interfragmentary wires, and pelvic alignment were assessed for associations with screw loosening. RESULTS: Complications developed in 96 of the 332 (29%) procedures. Cancellous screws without sacral engagement were associated with the lowest frequency (6%) of loosening, compared with cancellous and cortical screws engaging the sacrum and cortical screws that did not engage the sacrum. Frequency of screw loosening increased when cortical or cancellous screws engaged the sacrum and when cortical screws were used. In dogs that had surgery bilaterally, the first limb on which TPO was performed had a higher frequency of screw loosening than the second limb. Pelvic alignment loss was greatest (5.4 degrees ) when the 3 most cranial screws were loosened. Loss of pelvic alignment was significantly different between dogs that underwent surgery and had complications and those that underwent surgery and did not have complications in association with loosening of 1, 2, and 3 screws. CONCLUSIONS AND CLINICAL RELEVANCE: TPO screw loosening was multifactorial and related to stability of the affected ilium, screw type, and screw position. Placing cancellous screws that do not engage the sacrum in pelvic osteotomy plate positions 1 through 3 may decrease the number of screws that loosen.     

Effect of adrenalectomy on cholecystokinin-8-induced Fos-like immunoreactivity in myenteric neurons and the dorsal vagal complex in rats

OBJECTIVE: To investigate the effect of adrenalectomy on cholecystokinin-8 (CCK-8)-induced Fos-like immunoreactivity (Fos-LI) in the myenteric neurons of the dorsal vagal complex (DVC) in rats. ANIMALS: 16 male Sprague Dawley rats. PROCEDURES: Rats were allocated to 1 of 2 groups and underwent adrenalectomy or a sham adrenalectomy procedure. Rats were challenged with a supraphysiologic dose of CCK-8 (40 microg/kg) or physiologic saline (0.9% NaCl) solution (0.5 mL) administered IP; after 90 minutes, rats were euthanized, and Fos-LI was quantified in the DVC (at the levels of the area postrema, nucleus tractus solitarii, and dorsal motor nucleus of the vagus) and the myenteric neurons of the duodenum and jejunum by use of a diaminobenzidine reaction enhanced with nickel. The Fos-LI-positive cells were counted by use of an automated system and manually in the DVC and intestinal samples, respectively. Counts of Fos-LI in the different hindbrain levels and myenteric neurons were compared between the adrenalectomy--and shamtreated groups and between the CCK-8- and saline solution-treated groups. RESULTS: After adrenalectomy, CCK-8-induced Fos-LI was attenuated only in the myenteric neurons of the duodenum. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that the adrenal gland has a role in the activation of myenteric neurons by CCK-8 in rats.