Genotype-by-environment interactions at the trait level and total merit index level for milk production and functional traits in Brown Swiss cattle.

The production environments of the German-Austrian Brown Swiss population show a wide range due to differences in topography, landscapes, local climates, and different farm management systems. Extensive production systems such as organic farming have become increasingly popular in recent decades because of interest in sustainability and consumer preferences. Compared with conventional farmers, organic farmers put more weight on fitness traits. Besides the official total merit index (TMI), a selection index applying relative economic weights (REWs) suitable for organic production systems is provided for Brown Swiss cattle in Germany. The aim of the study was to investigate genotype-by-environment interactions (GxE) for milk production traits and functional traits (including longevity, fertility traits, and calving traits) in a sample of the German-Austrian Brown Swiss population housed in Baden-Wuerttemberg (southern Germany) by applying bivariate and random regression sire models. For bivariate analyses, the production environment was binary classified by farm management system (organic and conventional) and altitude of farm location (above or below 800 m above sea level (ASL)). Milk energy yields (MEY) obtained from herd effects were used as continuously scaled environmental descriptor in the reaction norm approach. The TMIs for sires were calculated based on breeding values estimated with different models and environment-specific REWs to determine possible GxE at TMI levels and rerankings of sires. In bivariate analyses, genetic correlations at the trait level were high and ranged from rg = 0.99 (calving to first insemination, cystic ovaries, and maternal stillbirth rate) to rg = 0.79 (first insemination to conception for altitude). Except for the latter, no severe GxE were found at the trait level using the bivariate models. Fat yield was the only trait showing minor GxE in the reaction norm model approach. Investigating the environmental sensitivity at the TMI level revealed rank correlations between the different environment-specific TMIs that were close to unity, implying no severe reranking effects. The results show no need to account for different environments in Brown Swiss cattle breeding programs.

Dissolution of solid lipid extrudates in biorelevant media.

Solid lipid extrudates with the model drug praziquantel were produced with chemically diverse lipids and investigated regarding their dissolution behaviour in different media. The lipids used in this study were glyceryl tripalmitate, glyceryl dibehenate, glyceryl monostearate, cetyl palmitate and solid paraffin. Thermoanalytical and dissolution behaviour was investigated directly after extrusion and after 3 and 6 months open storage at 40°C/75% RH. Dissolution studies were conducted in hydrochloric acid (HCl) pH 1.2 with different levels of polysorbate 20 and with a biorelevant medium containing pancreatic lipase, bile salts and phospholipids. Furthermore, the impact of lipid digestion on drug release was studied using in vitro lipolysis. The release of praziquantel from cetyl palmitate and glyceryl monostearate in the biorelevant medium was much faster than in HCl, whereas there was hardly any difference for the other lipids. It was shown that drug release from glyceryl monostearate matrices is driven by both solubilisation and enzymatic degradation of the lipid, whereas dissolution from cetyl palmitate extrudates is dependent only on solubilisation by surfactants in the medium. Moreover, storage influenced the appearance of the extrudate surface and the dissolution rate for all lipids except solid paraffin.

Effect of polymorphisms in four candidate genes for fertility on litter size in a German pig line.

We carried out an SNP discovery project in pigs for candidate genes playing potentially important roles in embryonic development. Using eight pigs one each from eight breeds (Meishan, Mangalitza, Duroc, Pietrain, German Landrace, Hampshire, Husum Red Pied, German Large White), 36 SNPs were identified in intronic sequences of 21 porcine candidate genes based on sequencing of PCR products. The primer pairs were designed using porcine EST sequences allowing amplification of introns. These SNPs were tested for their association with the number of piglets born alive in German Large White sows using a discordant approach. Significant effects (p < 0.001 and p < 0.05, respectively) of intronic SNPs on litter size were found for four genes: mitogen-activated protein kinase kinase kinase 3 (MAP3K3), vascular endothelial growth factor receptor (KDR), erbb2 interacting protein (ERBB2IP) and peroxisome proliferator-activated receptor delta (PPARD). These SNPs can be further tested in upcoming association studies for their influence on litter size in different breeds using larger sample sizes.

INHBA-associated markers as candidates for stallion fertility.

The inhibin beta A (INHBA) gene was chosen as candidate for stallion fertility and analysed for intragenic markers to find associations with pregnancy rate per oestrus. Intragenic single nucleotide polymorphisms (SNPs) were developed in order to perform an association and haplotype analysis using the least square means (LSM) of the pregnancy rate per oestrus for stallions as well as breeding values (BVs) for the embryonic and paternal component of the pregnancy rate per oestrus. The polymorphisms were genotyped in 161 Hanoverian warmblood stallions. Insemination records from approximately 20,000 Hanoverian warmblood mares were used to calculate LSM for stallions and to predict the paternal and embryonic component of BVs for the pregnancy rate per oestrus. We demonstrated significant associations of single markers and haplotypes with the LSM and the embryonic and paternal component of BVs for the pregnancy rate per oestrus. This is the first report on INHBA as an associated candidate gene with the LSM of stallions and the paternal and embryonic component of BVs for the pregnancy rate per oestrus.

Effect of polymorphisms in the genes for LIF and RBP4 on litter size in two German pig lines.

The association of two diallelic polymorphisms in the porcine genes for leukaemia inhibitory factor (LIF) and retinol-binding protein 4 (RBP4) with number of piglets born alive (NBA) in two German pig lines was studied. The investigated single nucleotide polymorphism (SNP) in the porcine LIF gene has been located in the 3'-untranslated region of its third exon, whereas the SNP in the RBP4 gene genotyped in this study is intronic. At the LIF locus the allele frequencies were 0.613 for the A allele and 0.387 for the B allele in German Landrace (GL) and 0.276 for A and 0.724 for B in German Large White (GW). At the RBP4 locus, the allele frequencies were 0.586 for the A allele and 0.414 for the B allele in GL and 0.733 for A and 0.267 for B in GW. There was a significant additive effect of the LIF B allele on NBA in GW over all parities (p

A polymorphism within the equine CRISP3 gene is associated with stallion fertility in Hanoverian warmblood horses.

Fertility of stallions is of high economic importance, especially for large breeding organisations and studs. Breeding schemes with respect to fertility traits and selection of stallions at an early stage may be improved by including molecular genetic markers associated with traits. The genes coding for equine cysteine-rich secretory proteins (CRISPs) are promising candidate genes because previous studies have shown that CRISPs play a role in the fertilising ability of male animals. We have previously characterised the three equine CRISP genes and identified a non-synonymous polymorphism in the CRISP1 gene. In this study, we report one non-synonymous polymorphism in the CRISP2 gene and four non-synonymous polymorphisms in the CRISP3 gene. All six CRISP polymorphisms were genotyped in 107 Hanoverian breeding stallions. Insemination records of stallions were used to analyse the association between CRISP polymorphisms and fertility traits. Three statistical models were used to evaluate the influence of single mutations, genotypes and haplotypes of the polymorphisms. The CRISP3 AJ459965:c.+622G>A SNP leading to the amino acid substitution E208K was significantly associated with the fertility of stallions. Stallions heterozygous for the CRISP3 c.+622G>A SNP had lower fertility than homozygous stallions (P = 0.0234). The pregnancy rate per cycle in these stallions was estimated to be approximately 7% lower than in stallions homozygous at this position.

Genetic analysis of three different classification protocols for the evaluation of elbow dysplasia in German shepherd dogs.

OBJECTIVES: Three different scoring systems for elbow dysplasia and its radiographic signs were genetically evaluated in 2645 German shepherd dogs. METHODS: An animal model was used to estimate heritabilities and additive genetic and residual correlations for the three scoring systems: ED-SV, which is recommended by the International Elbow Working Group; ED-LA, developed by Lang and others; and ED-TH, proposed by Tellhelm. RESULTS: The effects of sex, age at examination and the correlation between the two factors were significant for all three scoring systems. Heritability estimates (se) were 0.18 (0.04) for ED-SV, 0.11 (0.03) for ED-LA and 0.16 (0.04) for ED-TH. The additive genetic correlations among the different single criteria for elbow dysplasia and the different elbow dysplasia scores were between 0.68 and 0.98, except for the criteria ununited anconeal process and osteochondrosis dissecans of the trochlea humeri, which were mostly genetically negatively correlated to the other radiological criteria. CLINICAL SIGNIFICANCE: The elbow dysplasia scores were determined by two genetically different traits. The possibilities for selecting German shepherd dogs with respect to elbow dysplasia might be improved by taking into account these two traits in the prediction of breeding values.

Complex segregation analysis of canine hip dysplasia in German shepherd dogs.

Complex segregation analyses were carried out to clarify the mode of inheritance of canine hip dysplasia (CHD) in German shepherd dogs. Data were used from 8,567 animals examined for CHD from 20 families with three to four generations. The existence of a major gene in addition to polygenic gene effects was detected. In the present study, a mixed model with a dominant major gene effect seemed to be most probable for dichotomous encoding (0: dogs without signs of CHD; 1: dogs with borderline/slight to severe CHD). In addition, mixed major gene inheritance was shown for a binary trait where borderline was assigned to dogs scored free from CHD and for a trichotomously encoded trait (0: dogs without signs of CHD; 1: borderline CHD; 2: mild to severe CHD). Although only small frequencies were found for the unfavorable homozygotic genotype AA, the probability of the AB genotype was high in affected animals. Selection schemes to reduce the frequency of the allele A should therefore efficiently improve existing breeding programmes in German shepherd dogs.

Evidence of a new leukemia inhibitory factor-associated genetic marker for litter size in a synthetic pig line.

The association of a diallelic polymorphism in the leukemia inhibitory factor (LIF) gene with reproductive, growth, and carcass traits was studied in a German synthetic pig line. The diallelic SNP has been located in the 3'-untranslated region of the third exon of the porcine LIF gene. Information on 955 litter records from 273 genotyped sows was used in the analyses with respect to the number of piglets born alive. To identify possible pleiotropic marker effects, the growth and carcass traits ADG and backfat thickness were tested for associations with the SNP within the LIF gene in this population. At the LIF locus, the allele frequencies were 0.27 for the A allele and 0.73 for the B allele. There was an indication of an additive effect on the number of piglets born alive, and a significant dominance effect of the B allele was observed for first, second, and third to 10th parities (P = 0.044). The dominance effect for the first parity amounted to -0.73 +/- 0.36 (P = 0.047). No associations were detected between the marker alleles and the growth and carcass traits.

Genetic evaluation of elbow angles as predictors of elbow dysplasia in German shepherd dogs.

Elbow angles were measured in X-rays of both elbows to elucidate the usefulness of such data for selecting against elbow dysplasia (ED) in German shepherd dogs. These measurements record the size, proportions and alignments of the anconeal process (PA), the radius (RA), the olecranon (OL), and the ulnar trochlear notch (UL). The reference system for evaluating the information content of the measurements was the score for ED (ED-SV) as recommended by the International Elbow Working Group. Data from 2645 X-rayed dogs born from 1998 to 2001 in 1331 kennels were analysed by using residual maximum likelihood procedures to estimate heritabilities, additive genetic correlations and residual correlations. The pedigree file included 11,426 dogs and contained ancestors for up to six generations. ED-SV was significantly influenced by sex, by age within sex and by month of birth. The elbow angles were significantly influenced by the month and year of birth, X-ray positioning of the dog's forelimbs, angle of elbow flection, litter size and number of dogs X-rayed by the veterinary practitioners. The following heritability estimates were found: h2=0.18+/-0.04 for ED; h2=0.76+/-0.04 for OL; h2=0.52+/-0.05 for PA; h2=0.50+/-0.04 for UL; and h2=0.39+/-0.04 for RA. The additive genetic correlations of ED-SV with three elbow angles (OL, UL and RA) were close to zero. A higher additive genetic correlation to ED-SV was found only for PA, for which r(g)=0.31. The distributions of predicted breeding values for susceptibility to ED were not affected by regarding the elbow angles as additional traits in the multivariate prediction procedure. Measurements of elbow angles were thus shown to be unsuitable for use in selection programmes against ED in German shepherd dogs.

Effects of different artificial insemination techniques and sperm doses on fertility of normal mares and mares with abnormal reproductive history.

The effects of different artificial insemination (AI) techniques and sperm doses on pregnancy rates of normal Hanoverian breed mares and mares with a history of barrenness or pregnancy failure using fresh or frozen-thawed sperm were investigated. The material included 187 normal mares (148 foaling and 39 young maiden mares) and 85 problem mares with abnormal reproductive history. Mares were randomly allotted into groups with respect to AI technique (routine AI into the uterine body, transrectally controlled deep intracornual AI ipsilateral to the preovulatory follicle, or hysteroscopic AI onto the uterotubal junction ipsilateral to the preovulatory follicle), storage method of semen (fresh, frozen-thawed), AI volume (0.5, 2, 12 ml), and sperm dose (50 x 10(6) or 300 x 10(6) progressively motile sperm (pms) for fresh semen and 100 or 800 x 10(6) frozen-thawed sperm with >35% post-thaw motility). The mares were inseminated once per cycle, 24 h after hCG administration when fresh semen was used, or 30 h for frozen-thawed semen. Differences in pregnancy rates between treatment groups were analyzed by Chi-squared test, and for most relevant factors (insemination technique, mare, semen, and stallion) expectation values and confidence intervals were calculated using multivariate logistic models. Neither insemination technique, volume, sperm dose, nor mare or stallion had significant effects (P > 0.05) on fertility. Type of semen, breeding mares during foal heat, and an interaction between insemination technique, semen parameters, and mares did have significant effects (P < 0.05). In problem mares, frozen semen AI yielded significantly lower pregnancy rates than fresh semen AI (16/43, 37.2% versus 25/42, 59.5%), but this was not the case in normal mares. In normal mares, hysteroscopic AI with fresh semen gave significantly (P < 0.05) better pregnancy rates than uterine body AI (27/38, 71% versus 18/38, 47.3%), whereas in problem mares this resulted in significantly lower pregnancy rates than uterine body AI (5/15, 33.3% versus 16/19, 84.2%). Our results demonstrate that for problem mares, conventional insemination into the uterine body appears to be superior to hysteroscopic insemination and in normal mares, the highest pregnancy rates can be expected by hysteroscopic insemination.

Candidate gene markers for litter size in different German pig lines.

Three diallelic RFLP markers at candidate gene loci for litter size, the estrogen receptor (ESR) gene, the prolactin receptor (PRLR) gene, and the retinol-binding protein 4 (RBP4) gene, were evaluated for their association with the number of piglets born alive in different German pig lines. Genotyping was performed on boars and sows belonging to three different genetic groups from a single farm. Information on 8,336 litter records from 2,159 sows (German Landrace, n = 1,672; Duroc, n = 214; and a synthetic line, n = 273) was used in the analyses with respect to litter size. Growth performance traits were only analyzed for the synthetic line. The ESR locus showed no polymorphism in the tested boars of the German Landrace and Duroc lines. In the synthetic line, the frequency for the A allele was 0.90 and no homozygous BB animal was detected. No significant associations of ESR alleles with number of piglets born alive, backfat thickness, or average daily gain were observed. A new PCR-RFLP was developed for testing the PRLR polymorphism. The frequencies of PRLR allele A were 0.40 in the German Landrace, 0.49 in the synthetic, and 0.82 in the Duroc line. In the Duroc line, a small additive effect of the allele B on litter size was observed. The allelic substitution effect was 0.71 piglets born alive across all parities (P = 0.05). No significant associations of the PRLR locus with litter and growth performance traits were detected. The frequencies of RBP4 allele A ranged from 0.62 in the synthetic line to 0.67 in the German Landrace to 0.85 in the Duroc line. For the genotyped sows of the synthetic line, there was no indication of a favorable effect of the A allele with respect to litter size. Results of this study demonstrate that allele effects differ between lines or populations. This may be due to possible different linkage phases between the marker alleles and the causal mutations in the different lines. The results may also be explained by many minor genes affecting litter size. A selection strategy should be designed for each line separately and should always consider possible pleiotropic effects.

Attitudes toward the genetic testing of children among adults in a Utah-based kindred tested for a BRCA1 mutation.

Advances in molecular biology and genetics have led to the identification of the breast/ovarian cancer susceptibility genes BRCA1 and BRCA2, along with tests to detect mutations in these genes. Although the appropriateness of BRCA1/2 genetic testing for children has been debated in the literature, little is known about the attitudes of individuals who have undergone cancer susceptibility testing. The present study focused on attitudes toward BRCA1 testing for children among 218 adults from a Utah-based kindred who had received BRCA1 test results. Results indicated that approximately one-fourth of the participants would permit BRCA1 testing for children under the age of 18. General attitudes about genetic testing were predictive of attitudes toward the testing of children. In addition, men and individuals without a BRCA1 mutation were more likely to agree that minors should be allowed BRCA1 testing. Individuals whose mother had been affected with breast cancer were less likely to permit testing for minors. Among parents of minor children, less than one-fifth indicated that they would want BRCA1 testing for their own children; carrier status was not predictive of attitudes toward testing their own children. As breast/ovarian cancer susceptibility testing continues to be disseminated into clinical settings, there may be an increase in the number of test requests for minors. The findings of the present study represent an important step in exploring attitudes about genetic testing of children among individuals who have received cancer susceptibility test results.

[Preoperative determination of optimal amputation level in end-stage arterial occlusive disease]. / Die präoperative Bestimmung der optimalen Amputationshöhe im Endstadium der arteriellen Verschlusskrankheit.

For the preoperative determination of the amputation site in occlusive arterial disease of the lower extremity (i.e. healing by first intention in spite of a peripheral amputation level below the knee or at the knee) we developed a score by analyzing 500 patients with 523 amputations retrospectively. By means of a prospective study (100 consecutive lower limb amputations) this score has proved to be a helpful tool for realizing our therapeutic principle.

Indication for surgery of asymptomatic carotid lesions.

The incidence of cerebrovascular diseases with transient or persistent neurologic dysfunction has increased significantly. Although patients with symptomatic carotid artery stenosis clearly benefit from operative therapy, the indication to prophylactic surgery of asymptomatic carotid lesions however is still controversial. Based on data from a recently completed prospective randomized study and on analysis of the literature the indication and results of surgical treatment of asymptomatic stenoses of the carotid arteries are discussed. From 1970 to 1990 a total of 744 uni-or bilateral reconstructions of the internal carotid artery were performed in 631 patients. The perioperative morbidity (permanent neurologic deficiency) and mortality was 1.1% (n = 8) resp. 0.8% (n = 6). During the follow up period up to 18 years another 9 patients suffered from stroke (1.2%). The annual stroke incidence amounted to 0.2%. An important prerequisite for surgery is the so called critical internal carotid artery stenosis, implying reduced cerebral vasomotor reactivity or high embolic risk of an ulcerative plaque. Proper selection of patients (exclusion of multiple concomitant diseases) and an experienced team of vascular surgeons with operative morbidity and mortality below 1-2% validates surgical treatment of asymptomatic carotid artery stenoses.

[Para-tibial fasciotomy]. / Die paratibiale Fasziotomie.

Fifty patients with large venous ulcers have been surgically treated by paratibial fasciotomy plus dissection of incompetent perforating veins since January 1990. The operative mortality rate was 0.0%. A single significant complication (hematoma) required a secondary operation. Forty-eight ulcers healed within 8 weeks completely. In two cases, the venous ulcers recurred. Healing could be obtained under conservative treatment. Our results suggest that paratibial fasciotomy is the method of choice for the treatment of chronic deep venous insufficiency stage III after unsuccessful conservative treatment.

[Postoperative results of treatment in comparison with results of the German prospective randomized CASANOVA Study]. / Postoperative Behandlungsergebnisse im Vergleich zu den Resultaten der prospektiven randomisierten CASANOVA-Studie der BRD.

Independent of the results pending from the postoperative randomized German multicenter trial (CASANOVA Study), in which the course of operative versus non-operative treatment of asymptomatic carotid artery stenoses is being compared, there is still a clear justification for operative desobliteration if the following criteria are fulfilled: 1. Severe carotid stenosis with hemodynamic significance is proved by an extremely reduced or abolished autoregulatory reserve in the transcranial Doppler CO2-test. 2. Lesions with high embolic risk are confirmed by arteriography or B-Scan sonography. In 744 patients who underwent surgery according to these prerequisites there was a mere annual stroke rate of 0.2% during a followup of up to 18 years. This means that in comparison with the results of non-surgical therapy (annual stroke rate 1-3%) the risk of stroke was reduced by tenfold.

[Microbiological development studies in a clinically healthy, closed primate colony (Macaca mulatta, Macaca arctoides) after the establishment of a Salmonella infection]. / Mikrobiologische Verlaufsuntersuchungen in einer klinisch gesunden, geschlossenen Primatenkolonie (Macaca mulatta, Macaca arctoides) nach Feststellung einer Salmonellose.

A case of a Salmonella infantis septicemia in a primate (Macaca mulatta) probably induced by shipment stress gave rise to microbiological examination of fecal samples from all animals of a since 1976 closed colony of Macaca mulatta and Macaca arctoides. During 3 investigations carried out in 3 week intervals salmonella-positive animals were separated immediately from the stock. At the first examination 3 of 52, and at the second 2 of 49 samples were found to be salmonella-positive. At the third series all of the remaining 47 animals proved to be salmonella-negative. All isolates were identified as S. infantis. Feeding of contaminated, dried prawns was suspected as the most probable source of infection. A simultaneous examination of 96 of the stool specimens revealed Y. enterocolitica in 4 samples. Additionally, electron-microscopy of pooled fecal samples (3 to 4 animals each) led to demonstration of corona-, adenovirus and small round virus-like particles in one pool each. Moreover, coronavirus-like particles could be detected in 7 of 13 (1st examination series), 5 of 15 (2nd series) and 1 of 13 pools (3rd series).

[Incidence, causes and follow-up of doubtful smear findings (Papanicolaou group III)]. / Häufigkeit, Ursachen und Verlauf zweifelhafter Abstrichbefunde (Gruppe Papanicolaou III).

Suspect smears of 622 patients from 1977-1986 were analysed for their causes and followed up until the definite cytologic or histologic result. In relation to the overall material of 97,963 cases the rate of suspect smears (group Pap. III) was between 1.18 and 0.27%, i.e. an average of 0.68%. The most frequent reasons for the classification "suspect" were inflammatory (42.4%), metaplastic or degenerative changes (16.1%) and doubtful pre-cancer (30.6%). 58% of all cases became negative under cytological control and antimicrobial therapy or hormonal substitution; of these 91.4% converted into negative within 6 months. The histological examination carried out on 230 patients revealed pre-cancer (CIN I-III) in 201 cases; cancer was proved in 5 cases. Compared to the whole population that is a rate of 33.1%. 24 cases were to be found negative. 139 Multiparae (67.6%) were to be found in the group of the 206 histological positive cases. 151 women (74%) had taken hormonal contraceptives. The rate of suspect smears should not exceed one per cent.