Serotoninergic pathways are involved in economically important bovine gastrointestinal (GI) motility disorders such as displaced abomasum and cecal dilatation/dislocation. The existing research tools to investigate the role of serotoninergic pathways in such disorders in ruminants comprise functional pharmacological methods, e.g., in vitro contractility studies in tissue baths, and electromyographical recordings in vivo. However, no tools for quantification of bovine serotonin receptor [5-hydroxytryptamine receptor (5-HTR)] expression were available so far. This study aimed to develop real-time RT-PCR assays for quantitative mRNA analysis of bovine 5-HTR subtypes. Because the bovine 5-HTR coding sequences (CDSs) were completely unknown, multiple species (human, mouse, and rat) alignment of complete CDS was used for primer design in highly homologous regions. LightCycler real-time RT-PCR assays (partial CDS) for the following bovine 5-HTR subtypes were developed and validated: 5-HTR1A, 5-HTR1B, 5-HTR1D, 5-HTR1F, 5-HTR2A, 5-HTR2B, 5-HTR2C, and 5-HTR4. Intra- and inter-assay coefficients of variation (CV) for the eight established assays were small, ranging from 0.49% to 2.46%. As a first physiological application, 5-HTR mRNA expression levels were measured in brain, abomasum, and intestine of 10 healthy, lactating dairy cows. The 5-HTR expression was quantified by normalization to the housekeeping gene glyceraldehyde-phosphate-dehydrogenase (GAPDH). The 5-HTR subtype expression levels ranged from 0.001% (5-HTR2C in intestine) to 1% 5-HTR/GAPDH (5-HTR1B and 5-HTR4 in intestine). There were high variations of 5-HTR subtype mRNA expression within tissues across receptor subtypes and within receptor subtypes across tissues. In conclusion, accurate real-time RT-PCR assays for quantitative analysis of bovine 5-HTR subtype gene expression were developed and validated.
The objective of this study was to evaluate an investigative model which encompassed the risk factors, incidence, timing and causes of perinatal mortality (PM) (0 to 48h) on high risk dairy farms (PM of >5% in the previous year) in Switzerland. This pilot-study was carried out on 47 predominantly Holstein PM calves from 21 dairy farms, between September 2016 and January 2018.Gross pathological examinations of calves and placentae as well as histopathological examinations of internal organs and placental tissue were performed. Further investigations included microbiological examinations: broad-spectrum bacterial and fungal culture, detection of Chlamydia abortus, Coxiella burnetii, pathogenic Leptospira spp. and Neospora caninum by real-time PCR (qPCR) and of bovine viral diarrhoea virus (BVDV) by Ag-ELISA. Maternal blood samples were used for serology of bovine herpesvirus 1 (BHV-1), Brucella abortus, Chlamydia abortus, Coxiella burnetii and nine pathogenic leptospiral serovars and the evaluation of trace element status. A questionnaire was completed with the farmer, which included general farm characteristics and case-related data. Inbreeding coefficients (IC) were calculated for pure-bred matings.At the farm-level, the PM rate was 10.0% (5.3-28.2%) and at the cow-level, 11.5%. These values, from high-risk farms, were approximately five-times higher than the contemporary national bovine PM rate (2.3%) in Switzerland. The risk factors associated with these high PM rates were the self-selection of high risk herds, the high proportion of primiparae in these herds (45%) and the evidence of widespread pathogenic infections on these farms (exposure: 67% of herds, 53% of dams; infection: 57% of herds, 45% of calves). The majority (68.1%) of calves died intrapartum. The most commonly diagnosed initiating/ultimate cause of death (UCOD) was infection (34%) of which Coxiella burnetii was the most frequently detected pathogen, by antigen.The most frequently diagnosed proximate cause of death (PCOD) was asphyxia (44.7%), though multiple PCOD was also common (21.3%). This study was the first detailed investigation of bovine PM in Switzerland. Infectious causes were diagnosed more frequently than expected. While the findings from these high PM Swiss herds may have limited external validity, the investigative model adopted and the detailed research Journal Pre-proof 4 methodologies employed can be replicated and re-evaluated, respectively, in future studies on PM internationally.
Gentamicin-impregnated collagen sponges were used successfully in the treatment of chronic septic arthritis of the radiocarpal joint in two cattle. Both animals were moderately to severely lame and refractory to systemic antibiotics, and one of them was refractory to joint lavage and local antibiotics. The clinical diagnosis was confirmed by radiography and arthrocentesis. Arthroscopy was performed under general anaesthesia and, after debridement and lavage of the joint, gentamicin-impregnated collagen sponges were placed intra-articularly. Synovial fluid was sampled at 10 and 20 days after surgery and radiographs were taken three months (case 1) and two months (case 2) after surgery. The infection was eliminated from both animals and they recovered without residual lameness.
Prostaglandins are widely used in herd management due to their luteolytic properties. They have also a direct effect on the myometrium. We hypothesized, that dissimilar prostaglandin preparations would differ as to their contractile effect. Intrauterine pressure was recorded during the diestrus of lactating dairy cows using a transcervically placed intraluminal pressure microtransducer. After recording physiologic uterine motility for 30 min, prostaglandins (dinoprost, DL-cloprostenol, D-cloprostenol) or a placebo was administered intramuscularly, followed by a 2-h recording period. Significant differences were found for the area under the curve (P < or = 0.05) and mean amplitude (P < or = 0.05), whereas the number of spikes per 15 min and the baseline pressure during the last 3 min of every recording period did not differ significantly among treatments. Peak values for area under the curve and mean amplitude were found between 15 and 30 min after administration of DL-cloprostenol, while dinoprost yielded the steadiest plateau from this period until the end of the recording session. These results contrast with those of earlier studies comparing prostaglandins after intravenous administration.
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