Nuclear transfer in loach (Paramisgurnus dabryanus Sauvage) by cell-to-cell electrofusion

To study nuclear transfer in the loach (Paramisgurnus dabryanus Sauvage), blastula and gastrula cells were fused with UV-inactivated oocytes by cell-to-cell electrofusion. To facilitate nuclear transfer, blastula and gastrula cells were cultured or incubated at 4 °C in different solutions. TC-199 medium supplemented with 20% calf serum was the best culture solution, and effectively retained the totipotence of blastula or gastrula cells for up to 10 days. It was found that gastrula cells incubated at 4 °C had the same totipotence as blastula cells. The optimal UV dosage for inactivation of the oocyte chromatin was 180–240 mJ cm⁻². Electrofusion was carried out in a cone-shaped fusion chamber, which permitted the recipient oocyte and the donor blastula cell to contact one another. The electrofusion procedure resulted in a 10% success rate of normal-appearing fish. Genetic analysis indicated that the nuclear material originated from the donor cell (blastomere) and the oocyte pronucleus did not take part in development.     

Outcome of limb amputations in wapiti: 13 cases (1995-2001).

The object of this investigation was to determine the outcome of limb amputation in wapiti. Medical records of 13 wapiti that underwent limb amputation were reviewed to determine age, weight, sex, injury preceding amputation, limb amputated, amputation location, length of hospital stay, complications, and outcome. Ages ranged from 3 months to 8 years and weights from 70 kg to 280 kg. All animals were female. Eleven animals sustained catastrophic long bone fractures prior to amputation. Five animals had front limb amputations and 8 had hind limb amputations. Of these animals, 1 with a front limb amputaiton and 5 with hind limb amputations survived. Postsurgical inability to stand is associated with hospitalization of > 1 day. Of the 13 wapiti reviewed, 5 with hind limb amputations have produced and reared offspring. Hind limb amputation can be considered a viable alternative to euthanasia when catastrophic orthopedic injuries occur in female wapiti.     

A review of diet formulation strategies and feeding systems to reduce excretory and feed wastes in aquaculture

Significant efforts should be devoted to reducing waste outputs from aquaculture operations in order to lower the environmental impacts of aquaculture in many parts of the world. Since most aquaculture wastes are ultimately from dietary origin, reduction of waste outputs should first be through improvements of diet formulation and feeding strategies. The first step in the production of feeds producing less solid waste is to eliminate poorly digestible ingredients (such as whole grain or grain by-products used as binders and fillers in the feed formulae) and to use highly digestible ingredients with good binding properties. Further reduction of solid waste can then be achieved through careful selection of the ingredients to improve apparent digestibility and the nutrient balance of the feed. Nitrogen waste outputs can be reduced through the reduction of the digestible protein to digestible energy (DP/DE) ratio of the diet. Phosphorus waste outputs can be reduced through careful selection of the ingredients and optimization of the digestible phosphorus content of the diet to meet the requirement of the fish but avoid greatly exceeding this required level. Finally, feeding practices that minimize feed wastage should be adopted since feed wastage can have a very significant impact on waste outputs from fish culture operations.     

Genetic diversity estimates in Cicer using AFLP analysis

Amplified fragment length polymorphism (AFLP) analysis was used to evaluate the genetic variation among cultivated chickpea and wild Cicer relatives. In total, 214 marker loci were assessed, of which 211 were polymorphic (98.6%) across the 95 accessions that represented 17 species of Cicer. The genetic variation within a species was highest in C. pinnatifidum followed by C. reticulatum and lowest in C. macracanthum. Three main species groups were identified by UPGMA clustering using Nei's pair‐wise distance calculations. Group I included the cultivated species C. arietinum, C. reticulatum and C. echinospermum. Within this group, C. reticulatum accessions were clustered closest to the C. arietinum cultivars ‘Lasseter’, ‘Kaniva’ and ‘Bumper’, supporting the hypothesis that C. reticulatum is the most probable progenitor of the cultivated species. Cicer bijugum, C. judaicum and C. pinnatifidum were clustered together creating group II. Group III contained all nine perennial species assessed and two annual species C. yamashitae and C. cuneatum. The genetic distance detected between group I and group III (0.13) was equivalent to the genetic distance detected between group I and group II (the primary and annual tertiary species, respectively; 0.14). This indicated that the perennial tertiary species may be as valuable for increasing variation to incorporate novel germplasm in the cultigen as the annual tertiary species.     

Comparative evaluation of the effect of two maternal diets on fatty acids, vitamin E and carotenoids in the chick embryo.

1. The fatty acid profile of egg yolk and vitamin E and carotenoid accumulation in the egg yolk and embryonic tissues were investigated in relation to the maternal diet. 2. Two hundred fertile eggs (Ross 308 Broiler Breeder), obtained from hens fed on a maize-based (M-diet) or a wheat-based diet (W-diet), were incubated using standard conditions. 3. The egg yolk and embryo tissues (residual yolk, yolk sac membrane, liver, kidney, lung, muscles, adipose tissue and plasma) were collected on d 18 of incubation and on d 21 (newly-hatched chicks) and analysed for fatty acids, vitamin E and carotenoids. 4. The diets did not differ in terms of fatty acid or alpha-tocopherol concentrations. The concentration of carotenoids in the M-diet was 11.8 mg/kg and in the W-diet was 5.6 mg/kg with lutein and zeaxanthin being major carotenoids. 5. Eggs from the M-group contained higher (P<0.01) concentrations of beta+gamma-tocopherols, total carotenoids, lutein and zeaxanthin. Chickens hatched from those eggs were characterised by the increased concentrations of total carotenoids and zeaxanthin in all the tissues studied. The concentration of beta+gamma-tocopherol was enhanced only in the liver and yolk sac membrane. 6. It is concluded that the maternal diet plays an important role in antioxidant systems formation during chick embryonic development; the M-diet can increase the antioxidant potential of the egg yolk and embryonic tissues compared to the antioxidant potential provided by parent birds fed the W-diet.     

Enhanced recovery of avian influenza virus isolates by a combination of chicken embryo inoculation methods.

Since 1993, 14 cases of avian influenza from four different states in the U.S.A. have been diagnosed by virus isolation from eight avian species. Only 11 of the 14 avian influenza virus (AIV) primary isolations would have been successful if only the standard protocol for AIV isolation, i.e., inoculation of specific-pathogen-free embryonating chicken eggs (ECEs) by the chorioallantoic sac (CAS) route, had been followed. Primary isolation attempts were negative for AIV in three cases in which ECEs were inoculated by the CAS route; AIV could not be detected by hemagglutinating activity, agar gel immunodiffusion test or negative stain electron microscopy. However, in these three cases, primary isolations of AIV were achieved by inoculation of ECEs into either the yolk sac or onto the chorioallantoic membrane.     

Infectious disease serologic survey in free-ranging Venezuelan anacondas (Eunectes murinus).

Reptiles can harbor pathogenic microorganisms asymptomatically and serve as potential reservoirs of infection for humans, domestic animals, and other reptiles. Infectious diseases are also problematic for free-ranging reptile populations and are an important consideration in reptile reintroduction and translocation projects. There have been limited serologic studies of free-ranging reptiles for evidence of exposure to potential pathogens. In the present study, serum or plasma samples from five male and five female free-ranging Venezuelan anacondas (Eunectes murinus) were screened for antibodies to eastern, western, and Venezuelan equine encephalitis viruses, vesicular stomatitis virus, ophidian paramyxovirus, 19 Leptospira interrogans serovars, and Cryptosporidium serpentes. Antibodies to these agents were not detected, or antibody titers were low and possibly nonspecific. These results for the limited number of anacondas surveyed suggest that they do not serve as significant reservoirs for these infectious agents at this location.     

Toxoplasma gondii antibodies in exotic wild felids from Brazilian zoos.

Serum samples from 37 captive exotic felids in 12 zoos from six Brazilian states were assayed for antibodies to Toxoplasma gondii by the modified agglutination test using formalin-fixed whole tachyzoites. Titers greater than or equal to 1:20 were considered positive. Antibodies to T. gondii were found in 24 of 37 (64.9%) felids, including one European lynx (Lynx lynx), two jungle cats (Felis chaus), two servals (Leptailurus serval), two tigers (Panthera tigris), three leopards (Panthera pardus), and 14 of 27 lions (Panthera leo). This is the first serologic analysis for T. gondii infection in exotic wild felids from Brazilian zoos.     

Epidemiology and diagnosis of Mycobacterium tuberculosis in captive Asian elephants (Elephas maximus).

The deaths of two Asian elephants (Elephas maximus) in August 1996 led the United States Department of Agriculture to require the testing and treatment of elephants for tuberculosis. From August 1996 to September 1999. Mycobacterium tuberculosis infection was confirmed by culture in 12 of 118 elephants in six herds. Eight diagnoses were made antemortem on the basis of isolation of M. tuberculosis by culture of trunk wash samples; the remainder (including the initial two) were diagnosed postmortem. We present the case histories, epidemiologic characteristics, diagnostic test results, and therapeutic plans from these six herds. The intradermal tuberculin test, enzyme-linked immunosorbent assay serology, the blood tuberculosis test, and nucleic acid amplification and culture are compared as methods to diagnose M. tuberculosis infection in elephants.