Multiple independent de novo mutations are associated with the development of schistosoma reflexum, a lethal syndrome in cattle.

Schistosoma reflexum (SR) is a lethal congenital syndrome characterized by U-shaped dorsal retroflexion of the spine and exposure of abdominal viscera. SR is usually associated with severe dystocia. The syndrome is thought to be inherited as a Mendelian trait. We collected a series of 23 SR-affected calves from four breeds (20 Holstein, one Red Danish, one Limousin, one Romagnola) and performed whole-genome sequencing (WGS). WGS was performed on 51 cattle, including 14 cases with parents (trio-based; Group 1) and nine single cases (solo-based; Group 2). Sequencing-based genome-wide association studies with 20 Holstein cases and 154 controls showed no association (above Bonferroni threshold; P-value<3 ×10-09). Assuming a monogenic recessive inheritance, no region of shared homozygosity was observed, suggesting heterogeneity. Alternatively, the presence of possible dominant acting de novo mutations were assessed. In Group 1, heterozygous private variants, absent in both parents, were found in seven cases. These involved the ACTL6A, FLNA, GLG1, IQSEC2, MAST3, MBTPS2, and MLLT1 genes. In addition, heterozygous private variants affecting the genes DYNC1LI1, PPP2R2B, SCAF8, SUGP1, and UBP1 were identified in five cases from Group 2. The detected frameshift and missense variants are predicted to cause haploinsufficiency. Each of these 12 affected genes belong to the class of haploinsufficient loss-of-function genes or are involved in embryonic and pre-weaning lethality or are known to be associated with severe malformation syndromes in humans and/or mice. This study presents for the first time a detailed genomic evaluation of bovine SR, suggesting that independent de novo mutations may explain the sporadic occurrence of SR in cattle.

Decline of antibody response in indirect ELISA tests during the periparturient period caused diagnostic gaps in Coxiella burnetii and BVDV serology in pluriparous cows within a Holstein dairy herd.

In cattle, indirect enzyme-linked immunosorbent assay (ELISA) tests are commonly used in serological routine diagnostics. In a longitudinal study design, changes in relative optical density (OD) from drying-off until week 11 after calving were analyzed in blood and milk samples from pluriparous dairy cows (n=21) using a commercial indirect anti-C. burnetii ELISA test. In a second part of this study, changes over time were evaluated in blood and milk samples of 11 of these cows in an indirect ELISA detecting anti-Bovine viral diarrhea virus (BVDV) antibodies. Regarding the cows which were still in the herd at the time of calving, blood serological qualitative changes from positive to negative or indeterminate were demonstrated in 7/20 cows (35%) for the anti-C. burnetii indirect ELISA and in 2/10 cows (20%) for the anti-BVDV indirect ELISA, respectively, during the period from 14days ante partum to calving. Relative OD in the anti-C. burnetii and the anti-BVDV indirect ELISA tests followed basically similar courses over time. In blood serum, relative OD initially increased after drying-off, before a drop around calving was observed. After the colostrum period, relative OD in blood serum showed an increase until week 11 of lactation. In colostrum samples, relative OD levels were higher than in milk samples obtained one day before drying-off. After parturition, relative OD in milk decreased until week 6 of lactation in the anti-C. burnetii indirect ELISA and until week 11 in the anti-BVDV indirect ELISA, respectively. In conclusion, blood serological investigations in periparturient dairy cows using indirect ELISA kits should be avoided.

Rapid Discovery of De Novo Deleterious Mutations in Cattle Enhances the Value of Livestock as Model Species.

In humans, the clinical and molecular characterization of sporadic syndromes is often hindered by the small number of patients and the difficulty in developing animal models for severe dominant conditions. Here we show that the availability of large data sets of whole-genome sequences, high-density SNP chip genotypes and extensive recording of phenotype offers an unprecedented opportunity to quickly dissect the genetic architecture of severe dominant conditions in livestock. We report on the identification of seven dominant de novo mutations in CHD7, COL1A1, COL2A1, COPA, and MITF and exploit the structure of cattle populations to describe their clinical consequences and map modifier loci. Moreover, we demonstrate that the emergence of recessive genetic defects can be monitored by detecting de novo deleterious mutations in the genome of bulls used for artificial insemination. These results demonstrate the attractiveness of cattle as a model species in the post genomic era, particularly to confirm the genetic aetiology of isolated clinical case reports in humans.

Metritis vaccination in Holstein dairy heifers using a herd-specific multivalent vaccine - Effects on uterine health and fertility in first lactation.

In cattle, acute puerperal metritis (APM) compromises animal welfare and has an economic impact to the farmer because of the detrimental effects on reproductive performance, milk production, and survivability. The aim of this randomized clinical study was to investigate the effects of a prepartum immunization using a herd-specific multivalent vaccine on incidence of APM, prevalence of clinical endometritis (CE), and selected measures of reproductive performance in primiparous Holstein cows. Pregnant nulliparous cows (230±4days of gestation) assigned to the vaccinated group (VG; n=142) received subcutaneously 5.0mL of a multivalent herd-specific vaccine containing inactivated whole bacterial cells of Trueperella pyogenes, Escherichia coli, Streptococcus uberis, Bacteroides species, and Peptostreptococcus species obtained from uterine swabs of primiparous cows suffering from APM at the time of enrolment and three weeks later. Heifers allocated to the control group (Co; n=144) remained unvaccinated. Rectal temperature in the first 10days in milk (DIM) did not differ between treatments, and no interaction between treatment and day was observed (interaction treatment*day: p=0.623). Incidence of APM (VG, 46.0%, Co, 48.9%, p=0.588), number of antibiotic doses per cow to treat APM (VG, 0.63±0.81, Co, 0.64±0.75, p=0.496), prevalence of CE at 35 DIM (VG, 24.6%, Co, 19.3%, p=0.350), first service pregnancy per artificial insemination (VG, 40.3%, Co, 45.5%, p=0.541), number of inseminations until 150 DIM (VG, 2.0±1.4, Co, 1.9±1.3, p=0.749), interval from calving to first service (median days, VG, 52, Co, 52, p=0.915), and interval from calving to pregnancy (median days, VG, 90, Co, 83, p=0.419) did not differ between VG and Co. Treatment did not affect activity of aspartate aminotransferase (AST) and creatine kinase (CK) or concentrations of beta-hydroxybutyrate (BHB) and haptoglobin in serum samples collected at 5 DIM. Furthermore, treatment did not influence yields of milk, fat or protein in the first 100 DIM. Moreover, no signs that vaccination provided an attenuation of severity of the APM cases were detected as the course of rectal body temperature and values of AST, CK, BHB, and haptoglobin were not influenced by treatment within the group of cows with diagnosed APM. Use of this herd-specific multivalent vaccine designed to reduce metritis was not efficacious at reducing the risk of uterine disease or influencing production and reproduction in dairy cows.

Facets of Clinical Appearance and Aetiology in an Unusual Bovine Amorphus Globosus.

Amorphus globosus is a rare entity, more common in the cow but also reported in mares, buffaloes and goats. In respect of both development and clinical presentation, this abnormity can be evolved very variably. Previously, it has been discussed whether it is a form of twin pregnancy or placental teratoma. This case report deals with morphology and genetic observations in an unusual bovine amorphus globosus exhibiting a rudimentary clitoris and vulva, gut-like structures and rudimentary bones. The amorphus globosus was shown to be dizygotic to the normal male twin using the BovineSNP50 v2 BeadChip and had a genetical female sex. Aspects of aetiology and pathogenesis as well as the possible impact of amorphus globosus in the emergence of freemartinism are discussed.

Lack of effects of an equine chorionic gonadotropin (eCG) administration between days 9 and 15 postpartum on reproductive performance in a Holstein dairy herd.

Recently, it has been demonstrated that administration of equine chorionic gonadotropin (eCG) in the postpartum period in dairy cows can enhance follicle growth, reduce the interval from calving to first ovulation and increase plasma estradiol concentrations, and, thus, could enhance reproductive performance in a dairy herd when administered on day 6 postpartum. The aim of this study was to investigate the effects of a single dose of eCG between days 9 and 15 postpartum on parameters of reproductive performance in dairy cows. German Holstein cows (n = 1937; primiparous cows: n = 748; pluriparous cows: n = 1189) in a commercial dairy farm were randomly assigned to three experimental groups. Animals within the group eCG received a single dose of 600 IU eCG intramuscularly (i.m.) between days 9 and 15 postpartum followed by an i.m. administration of 500 µg cloprostenol after 14 days. Those of treatment group PG received cloprostenol only between days 23 and 29 postpartum. Cows of the control group remained untreated. Starting on day 49 postpartum, cows were subjected to a Presynch-Ovsynch protocol and inseminated artificially. The impact of application time (days postpartum) of eCG on the intervals calving to first service and calving to conception was statistically not significant. Outcomes of reproductive performance (i.e. first service conception rate, proportion of pregnant cows until 100 and 150 days in milk [DIM], number of inseminations until 150 DIM, calving to first service interval and calving to conception interval) did not differ significantly between treatment group eCG and group PG compared to control group. Regarding postpartum eCG administration, significant interactions between treatment and parity, season, milk yield, and early puerperal disorders, respectively, could not be shown. In conclusion, an eCG treatment of dairy cows between days 9 and 15 postpartum to increase reproductive performance cannot be recommended under the given circumstances.

Coxiella burnetii: Serological reactions and bacterial shedding in primiparous dairy cows in an endemically infected herd-impact on milk yield and fertility.

Coxiella burnetii (C. burnetii) is the causative agent of Q fever both in humans and animals. The objectives of this study were to investigate seropositivity and bacterial shedding in heifers and primiparous cows in an endemically infected herd and to assess the effects on post-partum diseases, fertility and milk production. At the age of 9 months, 96 Holstein heifers were included. Sampling was performed reproduction-orientated: at the beginning of the study, at detection of first pregnancy, 3 weeks before expected calving date (blood serum), at parturition and after 21, 42, 100 and 150 days in milk (DIM) (blood serum, vaginal swabs and milk). Serum samples were investigated by a commercial ELISA for the presence of specific antibodies and vaginal swabs and milk samples by PCR to detect C. burnetii DNA. Individual animal data (calving ease, stillbirth, retained foetal membranes, puerperal metritis, endometritis after 42 DIM, presence of corpus luteum after 42 DIM, interval calving-first service, interval calving-conception, number of inseminations until 150 DIM, proportion of pregnant cows until 100 and 150 DIM, proportion of pregnant cows after first service and data of the dairy herd improvement test) were documented. All heifers were seronegative at the age of 9 months and 3 weeks before the expected calving date. Subsequently, the proportion of seropositive animals and the antibody score increased significantly towards 42 and 100 DIM, respectively. Vaginal C. burnetii shedding was highest at parturition (30.9%), while the most positive milk samples were detected after 100 DIM (15.3%). Coxiella burnetii seropositivity and shedding had no impact on parameters of reproduction. However, milk fat yield was declined in puerperal vaginal shedders and cows which seroconverted during their first 42 DIM, respectively.

[Sporadic cutaneous lymphosarcoma of T-cell origin with involvement of lymph nodes and internal organs in a Holstein cow]. / Sporadische kutane T-Zell-Leukose mit Beteiligung von Lymphknoten und inneren Organen bei einer Holstein-Kuh.

Sporadic lymphosarcomas in adult cattle are rare entities with an unknown etiology. This case report describes the course of the disease in a 3.5-year-old cow of the breed German Holstein, which was presented to the veterinarian due to multifocal nodular skin lesions. Several superficial lymph nodes (Lymphonodi mandibulares, parotidei and mammariae) were enlarged, had a tight-elastic consistency and were freely movable. The histopathological and immunohistochemical examination of skin biopsies showed the presence of multifocal cutaneous T-cell lymphosarcomas consistent with a skin leukosis. Bovine leukemia virus infection was excluded by serological investigation of a milk sample and virological examination of a tissue sample, respectively. Seven weeks after the first clinical examination, the cow deteriorated rapidly and was euthanized. A post mortem examination revealed the presence of neoplastic cells within lymph nodes (all superficial lymph nodes of the carcass and Lymphonodi pulmonales), kidney and lungs as well as a liver rupture. Additionally, an overview of the case reports of sporadic bovine cutaneous lymphosarcomas published during the previous 15 years will be provided. The legal background for a further utilization of affected animals for milk and meat production will be discussed. This case report illustrates that sporadic bovine leukosis represents an important differential diagnosis for viral-, bacterial- and parasitic-induced skin lesions and enlargement of lymph nodes in adult cattle.

[Use of gonadorelin[6-D-Phe] at day 0 or 12 after insemination to increase the conception rate in a large dairy herd in Saxony/Germany]. / Einsatz von Gonadorelin[6-D-Phe] am Tag 0 oder 12 post inseminationem zur Steigerung der Konzeptionsrate in einer sächsischen Milchviehgroßanlage.

OBJECTIVE: To increase conception rates in lactating dairy cows by the application of a gonadotropin-releasing-hormone (GnRH)-agonist after insemination. MATERIAL AND METHODS: A total of 3125 inseminations of 1634 cows were included in this study. The animals were randomized into three groups at the time of insemination (day 0) using the final numeral of the ear-tag number: The cows of the GnRH0 group were treated using 100 µg gonadorelin[6-D-Phe] intramuscularly immediately after insemination (day 0), while those of the GnRH12 group were treated similarly at day 12. The cows of the control group received no hormonal treatment after insemination. An examination for pregnancy was performed at day 28 using transrectal ultrasonography. Analysis of the data sets was conducted for the number of inseminations (NI) 1-4 and for the last observed insemination of the respective cow during the experimental period, respectively. In a second step, statistical analysis was performed for the first service of cows with a lactation number of 1 versus > 1, with emphasis on the daily milk yield. In addition, a metritis diagnosed after the previous parturition was investigated as a possible factor influencing NI 1. RESULTS: Pregnancy risk at day 28 was decreased for NI 2 (n = 792) in the GnRH0 group compared to the control group (odds ratio [OR] = 0.69; 95% confidence interval [CI95] = 0.5-1.0; p = 0.04). A similar observation was found for NI 3 (n = 495) for the GnRH12 group (OR = 0.54; CI95 = 0.3-0.9; p = 0.01). In contrast, the pregnancy risk was increased for cows with a lactation number ≥ 2 and with a daily milk yield ≥ 42.5 kg (7-day-mean at day 0) at the first service (n = 364) by treatment with gonadorelin immediately after insemination (OR = 2.0; CI95 = 1.2-3.4; p = 0.01). No significant differences in the pregnancy risk were observed for the remaining analysed classes. CONCLUSION: An increased conception rate was only achieved for the first service of high-yield dairy cows (lactation number ≥ 2) by gonadorelin treatment at day 0. Presumably, the higher incidence of delayed ovulations in this group was treated successfully by gona- dorelin administration.

Transcoronary ablation of septal hypertrophy (TASH): a new treatment option for hypertrophic obstructive cardiomyopathy.

In 1991, our group started to develop a catheter interventional therapy for hypertrophic obstructive cardiomyopathy (HOCM). The new concept was proposed in 1994. It is based on the conventional PTCA technique with the aim of inducing an artificial myocardial infarction by instillation of 96% ethanol into the most proximally situated septal branch of the left anterior descending coronary artery. This leads to a subaortic contraction disorder with subsequent decrease of the intraventricular pressure gradient, shrinkage of the hypertrophied septal bulge and widening of the outflow tract ("therapeutic remodeling"). The subaortic defect is small and well demarcated as assessed by left ventricular angiography, transesophageal echocardiography and 18 F-glucose positron emission tomography. The term transcoronary ablation of septum hypertrophy (TASH) was suggested. Our patient cohort that now comprises 215 therapeutic procedures in 187 patients underwent a large variety of prospective studies (maximum follow-up 4.5 years) including invasive controls at regular intervals, investigation of hemodynamics at rest and at exercise, transesophageal and transthoracic echocardiography, Doppler echocardiography during bicycle exercise, electrophysiologic testing, Holter monitoring and measurement of myocardial metabolism and perfusion, assessment of microembolic events by transcranial Doppler sonography and histological examinations. This article gives an overview and reports our increasing experience in applying TASH. The following post-TASH findings were obtained: significant hemodynamic and clinical improvement at rest and at exercise, decrease of septum thickness, increase of outflow tract area and decrease of induced ventricular tachycardia. There were well-demarcated, histologically atypical subaortic myocardial defects, no microembolic events, abnormal early peak of infarct related enzymes, and no change of baroreflex sensitivity. Pre-/post-TASH evaluations of the patients should be based in particular on clinical symptoms correlated to the intraventricular gradient measured by bicycle exercise Doppler echocardiography and to outflow tract area as assessed by transesophageal echocardiography. Since 1994, as a roughly estimate, worldwide 1000 patients in 20 countries have been treated. According to published articles, abstract presentations and workshops, TASH consistently leads to a pronounced clinical and hemodynamic benefit for patients with HOCM. TASH has become an established technique. At least in centers with a high level of expertise, it is no longer experimental but a routinely performed alternative to surgical treatment for HOCM, i.e., the previous gold standard of therapy. Of course, patient outcome needs further careful clinical and prognostic evaluation. With respect to complications, TASH appears to be superior to surgery (transaortic septal myectomy) for HOCM. Like surgical treatment, TASH is currently indicated in critically ill patients with typical HOCM (subaortic form), who exhibit with drug refractory symptoms, including patients, who preferred DDD pacemaker therapy as a first therapeutic step but in whom this produced no subsequent clinical benefit.

Changes in the left ventricular outflow tract after transcoronary ablation of septal hypertrophy (TASH) for hypertrophic obstructive cardiomyopathy as assessed by transoesophageal echocardiography and by measuring myocardial glucose utilization and perfusion.

AIMS AND METHODS: Transcoronary ablation of septal hypertrophy (TASH) leads to marked clinical and haemodynamic improvement in patients with hypertrophic obstructive cardiomyopathy. In order to obtain more detailed information about changes in the outflow tract after TASH, transoesophageal echocardiography and a repeat invasive investigation were conducted before as well as 2 weeks and 6 months after TASH (n=62). In a subset of patients (n=11), metabolism and perfusion of the myocardium ((18)F-FDG-PET and(99m)Tc-MIBI-SPET) were investigated. RESULTS: After TASH there was a typical regional subaortic contraction disorder. It was quantified by a significant decrease in the fractional shortening of the left ventricular end-diastolic diameter, which declined from an average of 40.6% to 18.0%. The end-diastolic diameter increased from an average of 39.1 to 40.6 mm. There was also a significant reduction in septal thickness, which continued for up to 6 months after TASH, from an average of 20.0 mm to 11.1 mm in the region of ablation and from 23. 2 to 21.7 mm outside this region. The decrease in the gradient post TASH corresponded with a concomitant significant increase in the outflow tract area from a mean value of 1.04 cm(2)before the process to a value of 3.0 cm(2)after. In contrast to coronary heart disease, these changes were accompanied by non-diffuse, well demarcated subaortic-septal necrosis verified by(18)F-FDG-PET and(99m)Tc-MIBI-SPET. On average the TASH induced necrotic area comprised 6.6% of the left ventricle and correlated significantly with echocardiographic changes in the outflow tract. CONCLUSIONS: Alterations post TASH indicated that this catheter interventional treatment for hypertrophic obstructive cardiomyopathy affects the specific region of obstruction. The changes reflect a 'therapeutic remodelling' of the outflow tract of the left ventricle. They were demonstrable over the entire 6 months investigation period and obviously constituted the basis of post TASH clinical and haemodynamic improvement. Progressive alterations post TASH (post TASH reduction of subaortic septal thickness and an increase in the end-diastolic diameter) need special consideration during long-term follow up.