Combination of polyacrylamide gel electrophoresis with enzyme-linked immunosorbent assay: A simple method for determination of antibody specificity and detection of antigens in complex mixtures

Complex antigenic mixtures were separated on polyacrylamide gels in the presence of SDS. After electrophoresis, the gels were cut into equal fractions. Antigens were eluted from the fractions and could be bound to different solid phases on which a conventional enzyme-linked immunosorbent assay was performed. Antibody binding fractions could then be related with the bands of a stained gel run in parallel. This technique proved to be fast and sensitive. Antigens present in nanogram amounts in individual fractions were sufficient for the detection of specific antibody. We describe here the application of this technique to antibodies specific of parasite, viral, bacterial and mycoplasmal antigens and to antibodies against hormones and a muscle protein.     

Use of an immortalized bovine mammary epithelial cell line (MAC-T) to measure the mitogenic activity of extracts from heifer mammary tissue: effects of nutrition and ovariectomy

The objectives of the experiment were (1) to determine whether MAC-T cells would accurately mimic the previously observed proliferative responses of primary mammary epithelial cells (MEC) to mammary tissue extracts from high and low-fed heifers and (2) to determine whether mammary tissue extracts from ovariectomized (OVX) heifers would have lower mitogenic activity than intact controls. Addition of mammary tissue extracts to cell culture media of MAC-T cells plated on plastic or collagen-coated plastic to a range of concentrations between 1 and 8% resulted in dose-dependent increases in cell proliferation. Furthermore, mammary tissue extracts from low-fed prepubertal heifers aged 9 months, stimulated significantly more proliferation of MAC-T cells, as measured by 3H-thymidine incorporation into DNA than mammary tissue extracts from high-fed heifers (40.6 cpm x 10(3) per well versus 21.9+/-1.8 cpm x 10(3) per well). These observations suggested that MAC-T cells would be a suitable alternative to primary MECs for measuring the mitogenic activity of mammary tissue extracts. Conversely, no difference was observed in the mitogenic activity of mammary tissue extracts from OVX or control heifers. Possibly, MAC-T cells provide a good model for nutrition- but not ovarian-induced changes in mammary growth. Alternatively, that reduction of in vivo mammary development following OVX did not result in reduced mitogenic activity of the mammary tissue extracts emphasizes that heifer mammary development is the result of complex interactions between local growth factors and systemic hormones.     

Critical perspectives in animal agriculture: a response

People who work in the animal industries are faced with questions and criticisms about a variety of contentious issues, including animal management practices, ethics, diversity in animal agriculture, and animal welfare. Formulating responses to these questions requires a critical evaluation of our own work and open discussion of these controversial issues. Effective debate on these issues can be accomplished only with input from philosophers and social scientists skilled in such discussions, in addition to animal scientists. Therefore, animal scientists must engage in discussions of controversial issues among themselves and with entities outside agriculture. Furthermore, we must accept responsibility for the application of research results and any potential negative consequences. Because society is increasingly concerned with issues of animal welfare and the effects of new technologies, we should increase communications and transparency with the public. Increased diversity of race and gender will increase the ability of animal agriculture to connect with our stakeholders and to communicate the relevance of our work to society. Animal scientists need a professional ethic that espouses a higher level of understanding and commitment to philosophical discussions of contentious issues.     

The epizootiology of tuberculosis of cattle in the Federal Republic of Germany

Mycobacterial strains from different outbreaks of tuberculosis of cattle in Germany from 1996 to 2001 were differentiated by two molecular biological methods (Spoligotyping, RFLP IS6110). The causative agent was in one case Mycobacterium (M.) africanum, in 10 cases M. bovis and in 17 cases M. bovis ssp. caprae, respectively. The results of the molecular biological methods are discussed from the perspective of epizootiology and the particular importance of infections by M. bovis ssp. caprae emphasized. Direct contact of the animals, purchase from infected stocks, infected zoo animals and wildlife, as well as livestock handlers are discussed as possible sources of infection.     

Use of pneumocystoplasty for overinflation of the swim bladder in a goldfish

A Ryukin goldfish was evaluated because of a 6-month history of progressive abdominal distention and positive buoyancy. Overinflation of the swim bladder was diagnosed, and the fish was anesthetized with tricaine methanesulfonate. Archimedes' principle was used to determine the volume of swim bladder that was removed surgically. The caudal swim bladder was exteriorized through an abdominal incision and 2 surgical clips were placed across it to limit its size. After surgery, the fish remained in a state of negative buoyancy in sternal and lateral recumbency on the bottom of the tank. Sutures were removed 15 days after surgery, but the fish died 24 days after surgery. A full necropsy could not be performed because of autolysis of the tissues, but the surgical clips and the swim bladder appeared unremarkable. Pneumocystoplasty may be a viable treatment for this condition.     

Nymphal diapause and its photoperiodic control in the tick Ixodes scapularis (Acari: Ixodidae)

It is shown experimentally that the option between developmental diapause and non-diapause development in nymphs of Ixodes scapularis Say, 1821 (Middle Atlantic population) is determined by photoperiodic conditions according to a two-step photoperiodic reaction of short-day long-day type. Diapause arrest of development is induced by an impact of either long day upon unfed nymphs, or short day upon engorged nymphs, while non-diapause development completed in 2-2.5 months at 20 degrees C needs the change from short-day to long-day conditions. Some ecophysiological aspects of mechanisms controlling seasonal development of ticks belonging to Ixodes ricinus complex are discussed.     

Cytogenetic survey of Holstein bulls at a commercial artificial insemination company to determine prevalence of bulls with centric fusion and chimeric anomalies

OBJECTIVE: To determine prevalence of Holstein bulls with chromosomal anomalies, particularly the 1/21 centric fusion (CF), at a commercial artificial insemination (AI) company in the United States. DESIGN: Cross-sectional cytogenetic prevalence study. ANIMALS: All 606 Holstein bulls at a commercial AI company were cytogenetically screened to detect CF, chimerism, and other chromosomal abnormalities. PROCEDURE: Lymphocytes from heparinized blood samples were cultured by standard cytogenetic techniques, and chromosome spreads were prepared for microscopic examination. Chromosomal abnormalities were detected by examining 10 chromosome spreads per bull. Pedigree analysis was performed. RESULTS: None of the bulls had any type of CF. However, 6 bulls were identified as chimeras (i.e., contained lymphocytes with male [XY] and female [XX] chromosomes). One bull was sire or maternal grandsire to 85 of the bulls tested, and 739 of 1,212 (61%) sire and maternal-grandsire possibilities were accounted for by just 18 bulls. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these results supports previous indications that CF is extremely rare in Holstein bloodlines available commercially via AI in the United States. However, chimeric bulls are more common, and they reportedly have decreased reproductive performance. Therefore, identification of chimeric sires in the AI facility reported here and the possibility of de novo onset of CF at any time indicates that early cytogenetic screening should be encouraged for prospective bulls intended for use in AI programs.     

Ultrasonographic comparison of adhesions induced by two different methods of gastropexy in the dog

Ultrasonography was used to compare adhesions induced by two different methods of gastropexy in 16 dogs. An incisional gastropexy technique was used in eight dogs (group 1) and a 'modified' gastropexy technique in the remaining eight (group 2). The length and thickness of the gastropexy and the peristaltic activity of the stomach were measured ultrasonographically and compared between groups. Measurements for the two groups were taken in the early postoperative interval (two to four days), intermediate postoperative interval (eight to 20 days) and late postoperative interval (57 to 79 days). Both techniques were equally successful in forming permanent adhesions at two months postoperatively and there was no recurrence of gastric dilatation and volvulus. The length and thickness of the gastropexy were similar for both groups at two months postoperatively and there were no surgical complications with either technique. The modified gastropexy provides a technique that can easily be performed by a single surgeon , with no increased operative time or duration of anaesthesia, and with the formation of a permanent adhesion.