A multidisciplinary approach to determine factors associated with calf rearing practices and calf mortality in dairy herds.

In the Netherlands, an increase in ear-tagged calf mortality (3 days to 1 year of age) in dairy farms was observed. The aim was to determine why calf mortality increased and how to reduce calf mortality in herds with structural high rates. A multi-disciplinary approach was chosen to study this phenomenon. First analysis of census data revealed that the majority of the calves died in the first month of life. In addition, a panel of 236 farmers indicated that the increase in calf mortality might be related to priority, time management and the mind-set of farmers. For that reason a questionnaire was carried out to detect risk factors for mortality among young calves (<1 month) in 100 dairy farms with increased calf mortality compared to 100 dairy farms with stable and below average calf mortality. The results showed that, besides management factors such as IBR and BVDV control, and purchase of cattle, also the answers to statements giving an indication on the farmers' mind-set, were associated with calf mortality. Therefore, a qualitative sociological study on the farmers' identity was conducted by performing in-depth interviews among 30 farmers with structurally high calf mortality rates. Afterwards, the results were communicated with a veterinary advisor who visited the farmers and gave tailored advice. Most of the interviewed farmers believed to have sufficient knowledge and skills regarding calf rearing. The farmers did not share their calf rearing problems with colleagues and advisors but they mentioned to be open to receive advice if not communicated in a reproaching or pedantic way. The sociologist distinguished three different phases of awareness concerning calf mortality among the farmers: (1) farmers who were only partly, or not at all, aware of high calf mortality; (2) farmers who felt powerless because of their inability to find a solution to their problems; and (3) farmers who knew they can be inaccurate when it comes to rearing calves, but were reluctant to change this. With the background information of the farmers' identity it was easier for the veterinary advisor to provide tailored advice resulting in a higher probability of following up. A first evaluation in which calf mortality rates in the six months after providing the advice were monitored, indicated that the advice resulted in reduced mortality. The combination of census data, epidemiological and qualitative sociological research revealed that advisors should be aware of the attitude and mind-set of the farmer and adapt their approach and advice accordingly.

Effect of genetic variants of the heart fatty acid-binding protein gene on intramuscular fat and performance traits in pigs.

In order to find genetic markers to improve the meat quality of pigs by breeding we studied the relationship between variation in the heart fatty acid-binding protein (H-FABP) gene (FABP3) and intramuscular fat (IMF) content. To estimate the effect of H-FABP, pigs from two Duroc populations were selectively mated in such a way that at least two genotypes were present in each litter. In total, data from 983 pigs and pedigree information from three preceding generations were analyzed. Offspring were tested for IMF content as well as backfat thickness (BFT), BW, and drip loss of the meat (DRIP). All pigs were assigned to H-FABP RFLP genotype classes either by the assessed genotype (75%) or based on a probability score determined according to genotypic information of their relatives (25%). Contrasts were detected between homozygous H-FABP RFLP genotype classes for IMF content (.4%, P < .05), BFT (.6 mm, P < .01), and BW (2.4 kg, P < .10). No significant contrasts were detected for DRIP. Results for IMF content, BFT, and BW were confirmed when only genotyped animals were analyzed. Variation in BFT partially explained the effect on IMF content. Although other closely linked genes on porcine chromosome 6 might be responsible for the observed effect, interference of the halothane gene was excluded because all parental animals were noncarriers. In conclusion, H-FABP RFLP can be used as markers to select for increased IMF content and growth in breeding programs.

The adipocyte fatty acid-binding protein locus: characterization and association with intramuscular fat content in pigs.

The porcine A-FABP gene (FABP4) was isolated and sequenced to study the role of A-FABP in the differentiation of intramuscular fat (IMF) accretion in pigs. The coding sequence of the porcine A-FABP gene is highly conserved across human, mouse, and rat. Moreover, all the functionally important amino acids are conserved. This high similarity extends into the first 270 bp of the 5' upstream region. Within this region, a 56-bp nucleotide sequence was completely identical with the corresponding sequence in the mouse A-FABP gene, which contains the transcription factor binding sites for C/EBP and AP-1, and is implicated in the differentiation-dependent regulation of A-FABP. The A-FABP gene was assigned to porcine Chromosome (Chr) 4 by a porcine sequence-specific PCR on a cell hybrid panel, fully consistent with comparative mapping data with human and mouse. In the first intron of the porcine A-FABP gene, a microsatellite sequence was detected that was polymorphic for all six pig breeds tested. This genetic variation within the A-FABP gene was associated with differences in IMF content and possibly growth in a Duroc population, whereas no effect on backfat thickness and drip loss of the meat were detected. A considerable and significant contrast of approximately 1% IMF was observed between certain genotype classes. We conclude that the A-FABP locus is involved in the regulation of intramuscular fat accretion in Duroc pigs.

Identifying cytologic characteristics and grading endocervical columnar cell abnormalities. A study aided by high-definition television.

OBJECTIVE: To test the ability of cytotechnologists to recognize and accurately interpret selected architectural, cellular and nuclear features presented on a high-definition television (HDTV) and to make a reliable diagnosis with HDTV. STUDY DESIGN: A total of 1,122 features considered diagnostic of different endocervical columnar cell abnormalities were selected from 50 smears from 48 women with the help of a motor-driven-stage microscope by five observers who had knowledge of the final diagnosis. The selected and stored features were presented on an HDTV and evaluated in five successive sessions without knowledge of the final diagnosis. RESULTS: Specific types of features were correctly identified in a high number of cases. Considerable interobserver variability was demonstrated in the scoring of grades of expression of features. Overrated and under-rated monitor diagnoses were related to overvalued and undervalued features. From a group of 437 images that were correctly diagnosed by four or five observers, five features proved to be highly related to the correct diagnosis. CONCLUSION: Observers were capable of making a reliable diagnosis on features, selected by other observers, when presented on an HDTV. An overall correct diagnosis was made in 93% of cases.

Individual use of cytomorphologic characteristics in the diagnosis of endocervical columnar cell abnormalities: selection of preferred features with help of the 'Navigator' microscope.

In our previous interobserver studies on endocervical columnar cell abnormalities, we studied architectural, cellular and nuclear features in cervical smears of women known to have columnar cell atypias of variable severity, to determine cytomorphologic criteria, discriminating between mild, moderate and severe intraepithelial columnar cell lesions and adenocarcinoma. The results of these studies revealed a number of architectural, cellular and nuclear characteristics in different grades of expression, which were of importance for the primary diagnosis of: no abnormalities, different grades of intraepithelial endocervical columnar cell lesions and adenocarcinoma. Furthermore we concluded that observers used different characteristics and different grades of expression of these characteristics for comparable diagnoses. The present study was undertaken to determine those features, which were considered discriminating by each individual for the diagnosis of mild, moderate and severe atypia, adenocarcinoma in situ and adenocarcinoma. Features selected by five observers with knowledge of the final diagnosis, were stored and reviewed with help of a motor driven stage ('Navigator')-microscope and a high definition television-monitor. The results confirmed individual observer variability in the number and type of features used in the diagnosis of endocervical columnar cell abnormalities. Features such as 'variation in nuclear size and shape', 'irregular chromatin distribution' and 'coarsely granular chromatin' were selected preferentially by all observers in the diagnosis of endocervical columnar cell lesions, conversely striking differences were observed in the application of 'architectural'-and, especially in cases of 'adenocarcinoma', 'nucleolar' characteristics.

Endocervical columnar cell intraepithelial neoplasia (ECCIN). 3. Interobserver variability in feature use.

In our previous studies on endocervical columnar cell abnormalities, the ability of 5 cytotechnicians was tested to distinguish between cases of: no abnormalities, different grades of intraepithelial endocervical columnar cell atypias and invasive adenocarcinoma. On the basis of stepwise multiple regression analysis, nuclear chromatin distribution, variation in cellular and nuclear size, cytoplasmic eosinophilia and architectural features such as cell-crowding, cluster formation, formation of gland-like structures and pseudostratification appeared to be of primary diagnostic importance to discriminate between no abnormalities, different grades of cervical columnar cell atypias, adenocarcinoma in situ and adenocarcinoma. The present paper reports the results of a study which was designed to assess the individual performance in feature use and classification of endocervical lesions. The results of this study indicate that observers use different characteristics and different grading for the level of expression of these characteristics. However, all observers illustrated a strong relationship between the presence and the expression grade of a certain feature and the degree of endocervical columnar cell atypia. With acceptance of one grade of difference in grades of severity, an overall correct diagnosis was made in 87.6% of cases (range 80-98.2%).

CytosafePLUS. A workstation for screening, supervision, reviewing, quality assurance and education in cytopathology.

In pathology, the subjective cytologic and histologic diagnosis suffers from large observer variability. Effective methods of reducing the effects of this observer variability in cytologic diagnosis are close supervision of the diagnostic process and multiple screening by different observers of cases that have a higher risk of abnormalities as well as closely supervised follow-up procedures for early lesions. Accurate supervision of screening completeness has become possible with the development of computer programs and microscope instrumentation to register the path of the objective over the slide and therefore the fields that have been inspected by the screener (Axio-Home, Carl Zeiss, Oberkochen, Germany; Navigator, Becton Dickinson Cellular Imaging Systems, Leiden, the Netherlands). Based on these technologies, all objects of interest on the microscope slide are marked electronically, and the X,Y coordinates of these marks are stored in the computer. With these coordinates, a motor-driven microscope stage can be redirected to the selected objects, enabling the observer to quickly review the objects. The reviewer can thus reinspect cells or groups of cells, previously identified, in a much more accurate way than is presently possible, by marking objects of interest with ink dots. To these marked objects, specific diagnoses can be attached, as can comments or questions, which are also stored in the computer and are made part of the cytology report. After analysis, all marked objects can be relocated automatically with the help of the computer-directed microscope stage. Using this concept, a subsequent observer or a supervisor can easily and effectively reply to the remarks attached to these objects. It is also possible to reedit the comments made or to attach answers to specific questions. In an interobserver study, encompassing 50 cases of columnar cell abnormalities screened by five observers using the Navigator microscope stage and software, it proved possible to reduce reviewing time significantly. On the basis of marked cytologic characteristics discriminating cellular features in the diagnosis, endocervical columnar abnormalities could be identified. The coordinates of the objects of interest are made part of the laboratory record of the cytology findings (Cytosafe, Omnisys, Hoevelaken, the Netherlands). This development largely facilitates the review of previous findings when evaluating follow-up specimens. With the use of a Navigator motor-driven stage and CCD camera, microscope images can be made of the specimen under study. These images can be stored in the computer in a digitized format, labeled with their X,Y coordinates on the specimen. The CytosafePLUS workstation, encompassing the Navigator automated cell positioning system and CCD attachment, is a highly effective tool in cytologic screening, reviewing and reporting, in surveillance of follow-up, in intralaboratory and interlaboratory consultation, in cytology training, in proficiency testing and in quality assurance.

Endocervical columnar cell intraepithelial neoplasia. II. Grades of expression of cytomorphologic criteria.

OBJECTIVE: To determine the relationship between the grade of expression of discriminating cytomorphologic architectural, cellular and nuclear features and the degree of endocervical columnar cell atypia. STUDY DESIGN: The study group consisted of 55 smears from 53 women known to have endocervical columnar cell atypias of variable severity. The smears were reviewed by five observers by means of 46 architectural, cellular and nuclear features. Seventeen of the 46 evaluated features were scored as revealing three grades of expression: slight, moderate and marked. RESULTS: Grades of expression of cellular and nuclear characteristics used in the evaluation of endocervical columnar cell abnormalities were identical to the characteristics used in the diagnosis of squamous (metaplastic) lesions. As such the features were extremely useful, with the exception of hyperchromasia, which was less prominently expressed. Furthermore, the results on architectural features revealed that the expression grades slight and moderate were indicative of mild and moderate endocervical columnar cell atypia, whereas the marked expression grade was strongly related to the severe intraepithelial lesions and adenocarcinoma. CONCLUSION: Characteristic cytomorphologic features at different grades of expression proved to be reliable light microscopic parameters in the diagnosis of different grades of endocervical columnar cell abnormalities. The accuracy of diagnosis at least equaled that of detecting squamous (metaplastic) lesions.

Endocervical columnar cell intraepithelial neoplasia. I. Discriminating cytomorphologic criteria.

OBJECTIVE: To identify specific cytomorphologic characteristics of mild, moderate and severe atypia of endocervical columnar cell lesions. STUDY DESIGN: The study group consisted of 55 smears from 53 women known to have columnar cell atypias of variable severity. The smears were reviewed by five observers in terms of 46 architectural, cellular and nuclear features. Review diagnoses were given together with the scoring for a large number of cellular characteristics. RESULTS: Nuclear chromatin distribution, variation in cellular and nuclear size, cytoplasmic eosinophilia and such architectural features as cell crowding, cluster formation, formation of gland-like structures and pseudostratification appeared to be of primary diagnostic importance for discriminating between no abnormalities, different grades of severity of endocervical intraepithelial neoplasia and adenocarcinoma. With acceptance of one grade of difference in grades of severity, a correct diagnosis was made in 87.6% of cases. CONCLUSION: With the application of specific architectural, cellular and nuclear features, the cytologic detection of different grades of endocervical intraepithelial neoplasia and of adenocarcinoma proved to be dependable and reproducible, with accuracy that at least equaled that of detecting squamous (metaplastic) lesions.