A new single nucleotide polymorphism in CAPN1 extends the current tenderness marker test to include cattle of Bos indicus, Bos taurus, and crossbred descent

The three objectives of this study were to 1) test for the existence of beef tenderness markers in the CAPN1 gene segregating in Brahman cattle; 2) test existing CAPN1 tenderness markers in indicus-influenced crossbred cattle; and 3) produce a revised marker system for use in cattle of all subspecies backgrounds. Previously, two SNP in the CAPN1 gene have been described that could be used to guide selection in Bos taurus cattle (designated Markers 316 and 530), but neither marker segregates at high frequency in Brahman cattle. In this study, we examined three additional SNP in CAPN1 to determine whether variation in this gene could be associated with tenderness in a large, multisire American Brahman population. One marker (termed 4751) was associated with shear force on postmortem d 7 (P < 0.01), 14 (P = 0.015), and 21 (P < 0.001) in this population, demonstrating that genetic variation important for tenderness segregates in Bos indicus cattle at or near CAPN1. Marker 4751 also was associated with shear force (P < 0.01) in the same large, multisire population of cattle of strictly Bos taurus descent that was used to develop the previously reported SNP (referred to as the Germplasm Evaluation [GPE] Cycle 7 population), indicating the possibility that one marker could have wide applicability in cattle of all subspecies backgrounds. To test this hypothesis, Marker 4751 was tested in a third large, multisire cattle population of crossbred subspecies descent (including sire breeds of Brangus, Beefmaster, Bonsmara, Romosinuano, Hereford, and Angus referred to as the GPE Cycle 8 population). The highly significant association of Marker 4751 with shear force in this population (P < 0.001) confirms the usefulness of Marker 4751 in cattle of all subspecies backgrounds, including Bos taurus, Bos indicus, and crossbred descent. This wide applicability adds substantial value over previously released Markers 316 and 530. However, Marker 316, which had previously been shown to be associated with tenderness in the GPE Cycle 7 population, also was highly associated with shear force in the GPE Cycle 8 animals (P < 0.001). Thus, Marker 316 may continue to be useful in a variety of populations with a high percentage of Bos taurus backgrounds. An optimal marker strategy for CAPN1 in many cases will be to use both Markers 316 and 4751.     

Preliminary study of ethylene oxide sterilization of full-thickness cortical allografts used in segmental femoral fracture repair

Full-thickness canine cortical allografts were cleanly harvested, sterilized with ethylene oxide, and stored at room temperature. The allografts were incorporated into canine segmental femoral fracture repairs and compared clinically, radiographically, and morphologically with control femoral cortical autografts for function of the limb, graft acceptance, and bone union. Sterility was maintained and the cortical allografts were well accepted by the host animals, resulting in full use of the limb which was subjected to surgical operation. The allografts showed healing patterns similar to those of the autografts, as determined by radiographic, gross, and histologic evaluation of the proximal and distal host-graft interfaces. Evaluations were made monthly. The host-graft interfaces of the allografts and autograft were filled with woven bone with adjacent vascular invasion and remodeling of the graft at the final 4th-month evaluation.     

A luteinizing hormone-releasing hormone-induced serum luteinizing hormone surge is not detectable in the milk of cows

Six lactating Holstein cows were used to determine whether a serum luteinizing hormone (LH) surge induced by luteinizing hormone-releasing hormone (LHRH) could be detected in milk. A double antibody radioimmunoassay was evaluated for measuring LH in whole milk. Cows (d 10 of the estrous cycle) were injected with saline (time zero), followed by LHRH 12 h later. Blood samples were collected hourly for 12 h via jugular cannula following each injection; milk removal was accomplished every 2 h by a portable milking machine. On d 10 of the next estrous cycle, treatment, order was switched, with the same cows receiving LHRH at time zero and saline 12 h later. Approximately 2 h following LHRH treatment, serum LH levels peaked at 29 ng/ml and remained elevated for 5 h. There was no corresponding change in milk LH detected during the 12-h to 24-h period following the induced serum LH surge. Our conclusion is that the measurement of LH in the milk of cows shows little promise for predicting ovulation time in the cow.     

Bovine NK cells acquire cytotoxic activity and produce IFN-gamma after stimulation by Mycobacterium bovis BCG- or Babesia bovis-exposed splenic dendritic cells

Early interactions of innate immune cell populations, such as dendritic cells (DC) and natural killer (NK) cells, can affect the ability of the acquired immune response to control infection of intracellular microorganisms. In this study, we investigated the activation of bovine NK cells by CD13(+) splenic DC stimulated with either Mycobacterium bovis BCG or Babesia bovis merozoites. Splenic DC were used either immediately after selection (cytokine(-)) or after exposure to GM-CSF, IL-4 and Flt3L for 72 h (cytokine(+)). Phenotypic analyses showed up-regulation of MHCII, CD80 and CD86 on cytokine(+) DC when compared to cytokine(-) DC. Purified NK cells (CD335(+)CD3(-)CD2(+/-)CD8alpha(+/-)) were co-cultured with microbial-exposed cytokine(-) DC or cytokine(+) DC in either transwell or cell-to-cell format and NK cell IFN-gamma production and cytotoxicity were assessed. NK cell IFN-gamma production was dependent on cell-to-cell contact. Microbial-stimulated cytokine(+) DC induced significantly more IFN-gamma production from NK cells than cytokine(-) cells. In contrast, cytotoxicity and perforin up-regulation were more pronounced in NK cells cultured with cytokine(-) DC than cytokine(+) DC. Therefore, activation of bovine NK cells by microbial-stimulated CD13(+) splenic DC is influenced by the maturation state of the DC suggesting different roles for the splenic DC during disease-induced maturation.     

Application and Commercialization of Flow Cytometrically Sex-Sorted Semen

The current technology to sort X and Y chromosome bearing sperm population requires individual identification and selection of spermatozoa in a modified high-speed flow cytometer. For farm animal species, the technology is capable of producing sexed sperm at greater than 90% purity. However, only in the bovine, the technology has reached a developmental level that allows its commercial application. Meanwhile, the demand for female calves has grown rapidly, which encourages the demand for sex-sorted semen from high genetic value bulls. The success of the technology will depend mainly on the fertilizing capacity of the sorted spermatozoa, as this is the most affecting and economically relevant factor. To date, fertility is still variable and is quite dependant on post-sort processing. New processing techniques are under investigation and will likely be able to improve the fertility rates after AI with sex-sorted semen. It is of great importance to select the right bulls and to test the sorted samples on a routine basis. In addition to the demand for sex-sorted semen by the cattle industry, there is also a significant demand expressed by pig farmers. However, it is still unknown if the use of sex-sorted semen through commercial pig AI will be economically feasible. For the pig, the combination of in vitro fertilization with sexed semen and non-surgical embryo transfer is an alternative that merits further scientific attention. Recent developments in ovine AI and ET will make it very likely that commercial sheep industry will adopt the sexing technology in their breeding concepts.     

Udder edema in cattle: effect of furosemide, hydrochlorothiazide, acetazolamide, or 50% dextrose on venous blood pressure

Before injection of diuretic drugs, blood pressures were determined in the cranial superficial epigastric veins (milk vein) and jugular veins of 15 Holstein cows with udder edema at parturition and of 15 healthy (control) cows at parturition. Cows with udder edema had a significant (P less than or equal to 0.05) mean increase in cranial superficial epigastric venous pressure at parturition, compared with that of control cows. After IV administration of 500 mg of furosemide in cows with udder edema, the mean cranial superficial epigastric venous blood pressure significantly (P less than or equal to 0.05) decreased within 5 minutes, remained at the decreased pressure for 90 minutes, and then returned to near pretreatment pressures by 210 minutes after furosemide injection; however, furosemide did not significantly affect the jugular venous blood pressure of these cows. Furosemide injection did not significantly affect the cranial superficial epigastric or jugular venous blood pressure in the control cows. After IV administration of hydrochlorothiazide (250 mg), acetazolamide (500 mg), or 50% dextrose (500 g), the cranial superficial epigastric and jugular venous blood pressures in cows with udder edema at parturition were not significantly different from those in cows without udder edema at parturition during the 210-minute evaluation period after injection of the drugs.     

In vitro exposure of bovine morulae to Ureaplasma diversum.

Ureaplasma diversum has been associated with infertility in the cow experimentally and in naturally occurring cases. However, the pathogenic mechanism is undetermined. The purpose of this study was to determine whether ureaplasmas are pathogenic for bovine morulae in vitro. Twenty-one morulae were recovered from three superovulated, mature, Holstein cows six or seven days postestrus. The embryos were divided into three groups (A,B,C) and incubated for 16 hours at 37 degrees C in humidified air with 10% CO2. Group A was incubated in embryo culture medium alone, Group B was incubated in culture medium with sterile ureaplasma broth added and Group C was incubated in culture medium containing 1.7 X 10(6) colony forming units Ureaplasma diversum strain 2312. After incubation, the morulae were examined using an electron microscope. Structures morphologically identical to U. diversum were present on the outer surface of the zonae pellucidae of all the morulae exposed to the organism and none were present on the unexposed control embryos. No other morphological differences were observed in either the ureaplasma-exposed embryos or the two groups of control embryos. Ureaplasma diversum was isolated from three of the five embryos incubated in culture medium with sterile ureaplasma broth added. These three embryos were recovered from one donor cow which cultured positive for U. diversum from the vulva and flush fluid. This finding suggests that the contaminating organisms entered the embryo culture wells either in the embryo collection medium or attached to the embryos.(ABSTRACT TRUNCATED AT 250 WORDS)     

Visceral leishmaniasis in the German shepherd dog. II. Pathology

Three German shepherd dogs were inoculated with Leishmania chagasi and three with Leishmania donovani and the infection was followed for 82 days. All infected dogs developed splenomegaly and lymphadenomegaly. In lymph nodes there was a reduction in lymphocyte population in paracortical areas, extensive proliferation of macrophages in paracortical areas and medullary cords, follicular hyperplasia, and increased numbers of plasma cells. The spleen had decreased numbers of lymphocytes in periarteriolar lymphoid sheaths, proliferation of macrophages in these regions, follicular hyperplasia, and enlargement of the red pulp with clusters of macrophages and plasma cells. The morphology of the tonsil was similar to the lymph nodes. Clusters of macrophages, often containing Leishmania spp, were present in liver, bone marrow, lung, and the intestines. The morphologic changes in lymph nodes and spleen were suggestive of a suppressed cell-mediated immunity and an active humoral immunity. The German shepherd dog may be a useful laboratory model for the study of immunopathologic changes in visceral leishmaniasis.     

MULTICENTRIC LYMPHOMA WITH INTESTINAL INVOLVEMENT IN A DOG

A 5-year-old, male German shepherd dog was evaluated for a 5-week history of vomiting, diarrhea, lethargy, and weight loss. In abdominal radiographs, there was an unusual corrugated appearance within a loop of intestine. Sonographically, this loop also had a similar corrugated appearance with alternating hyperechoic and hypoechoic bands. Lymph nodes within the abdomen and thorax were enlarged, and a thoracic mass was also detected. Ultrasound-guided fine-needle aspirates of the mesenteric lymph nodes were highly suggestive of lymphoma. After postmortem examination, a diagnosis of multicentric lymphoma with involvement of the gastrointestinal tract was made.