Tank culture of yellowfin tuna, Thunnus albacares: developing a spawning population for research purposes

A land-based culture facility for research on yellowfin tuna, Thunnus albacares, was developed at the Achotines Laboratory in the Republic of Panama. Six concrete tanks, and seawater and life support systems were built to maintain a yellowfin broodstock. On average, 50% of the yellowfin caught survived capture and handling, and approximately 30% became broodstock in Tank 1 (17 m diameter, 6 m depth) or Tank 2 (8.5 m diameter, 3 m depth). Each fish was tagged with a microchip implant tag, then weighed, measured, and injected with oxytetracycline (OTC) prior to stocking. Daily rations of primarily market squid, Loligo opalescens, and Pacific thread herring, Opisthonema spp., were regulated based on the feeding activity and energy requirements of the fish. Feeding activity of the broodstock decreased when the water temperatures decreased, and the fish ate decreasing daily rations and increasing calories with increasing size. Spawning occurred in both tanks within 6–8 months of capture. Spawning first occurred in Tank 1 when 24 females ranged in size from 6 to 16 kg and 65 to 93 cm fork length (FL). Spawning was intermittent during the first 2 months and occurred near daily thereafter. Tank size appeared to affect survival rates, the types of mortalities that occurred, and the growth of the fish. Survival rates after 1 year in captivity were higher, and the fish were larger, on average, in Tank 1 than in Tank 2. Most of the mortalities in Tank 1 were the result of wall strikes, which occurred more frequently after the fish reached their highest density of 0.64 kg m⁻³ and sizes greater than 96 cm FL and 19 kg. Non-linear growth models were fitted to the initial stocking sizes and final sizes of fish that died or were removed from Tank 1 during 1996–1999. Estimated growth rates in length (11–48 cm year⁻¹) for fish between 51 and 150 cm FL decreased with increasing length. Estimated growth rates in weight ranged from 9 to 19 kg year⁻¹ for fish less than 19 kg and 20–23 kg year⁻¹ for fish greater than 19 kg. The results of this work demonstrate that the stable environment of a land-based culture facility may be the preferred system for long-term maintenance of a yellowfin broodstock.     

Metabolism of bromoxynil octanoate in growing wheat

[¹⁴C]ring-Bromoxynil octanoate was applied to the leaves of wheat seedlings, which were cultivated in a growth cabinet under controlled conditions for 14 days. Fractionation of the metabolites present in the treated leaves, which accounted for about 63% of the radioactivity applied, indicated a complex metabolic pathway resulting from initial hydrolysis to free bromoxynil, followed by three consecutive or concurrent steps (a) hydrolysis of the cyano group to the amide and carboxylic acid, followed by decarboxylation to 2,6-dibromophenol (0.5% of the ¹⁴C applied), (b) replacement of one or both bromine atoms by hydroxy groups to 3-bromo-4,5-dihydroxybenzonitrile (1.3 %) and 3,4,5-trihydroxybenzo-nitrile (0.6 %) or their hydrolysis products, (c) replacement of one or both bromine atoms by hydrogen, giving 3-bromo-4-hydroxybenzonitrile (1.9 %) and 4-hydroxy-benzonitrile (0.6%) or their hydrolysis products. Some of the phenolic acids or phenols formed are natural plant constituents. The metabolites identified represented in all about 11 % of the herbicide applied, but no individual metabolite accounted for more than a small proportion of it.     

Radiochemical distribution and decline studies with bromoxynil octanoate in wheat

Bromoxynil octanoate labelled with ¹⁴C in the ring or in the cyano-group was applied to wheat seedlings at the two-leaf or fully-tillered stage and at rates equivalent to up to 16 oz a.i./acre. The plants were grown either in environmental chambers under controlled conditions for up to 28 days, or outdoors under field conditions for various periods up to harvest. Initially, elimination of radioactivity occurred more rapidly with bromoxynil-cyano-[¹⁴C]-octanoate than with bromoxynil-ring-[¹⁴C]-octanoate, indicating metabolic attack on the cyano group. Under outdoor conditions with ring-[¹⁴C]-herbicide applied at the two-leaf stage, only 12% of the radioactivity was retained after 28 days, principally in the treated leaves. When application was made at fully-tillered stage, about 33% of the ¹⁴C was retained after 56 days, almost entirely in the treated senescent leaves at the base of the plant. There was very little translocation of the herbicide or of any major metabolite. The level of radioactivity in harvested grain and in straw more than 7.5 cm above the ground was very low, even after very late application of ring-[¹⁴C]-labelled herbicide. The amount of bromoxynil octanoate, together with any metabolite retaining part of the aromatic ring, did not collectively exceed the equivalent of approx. 0.01 parts/million bromoxynil octanoate.     

Diagnostic implications of detection of proteinase K-resistant protein in spleen, lymph nodes, and brain of sheep.

Brain, spleen, and selected lymph nodes from sheep with clinical signs of scrapie were analyzed for presence of proteinase K-resistant protein (PrP-res). Diagnosis of scrapie on the basis of detection of PrP-res was compared with diagnosis on the basis of histologic evaluation of the brain from clinically affected or exposed sheep. Proteinase K-resistant protein was found in every brain that was histologically positive for scrapie, and in addition, was found in the brain of several clinically positive sheep that were not diagnosed as scrapie-positive by histologic evaluation. Proteinase K-resistant protein was also found in 87% of the spleens and lymph nodes from sheep that had PrP-res detected in brain homogenates. Therefore, analysis of sheep brain, spleen, or lymph nodes for PrP-res provided a diagnostic approach that was superior to histologic examination alone for detection of naturally scrapie agent-infected sheep.     

Detection of colonies of Anaplasma marginale in salivary glands of three Dermacentor spp infected as nymphs or adults

Salivary glands from males of 3 Dermacentor species (D andersoni, D variabilis and D occidentalis) that were infected with either the Virginia or Idaho isolate of Anaplasma marginale as nymphs or adults were examined for colonies of A marginale by use of light and electron microscopy. Prior to dissection of salivary glands, exposed ticks were held at 25 C for 15 to 18 days, followed by a 3-day incubation at 37 C. Ticks of 2 species transmitted A marginale to calves; the third tick species was confirmed infected by demonstration of typical colonies in tick gut cells, but transmission was not attempted; Colonies of A marginale were seen with light microscopy in salivary glands of all 3 species of ticks; they were located in acinar cells that contained simple granules. Colonies varied morphologically from small, compact ones to larger structures that contained distinct organisms and often were adjacent to the host cell nucleus. Electron microscopy confirmed that the colonies were rickettsial organisms. Morphologic features of A marginale varied and included reticulated forms, forms with electron-dense centers, and small particles; these various forms were similar to those described previously in midgut epithelial cells of ticks. We believe that the organism seen within tick salivary glands may replicate in the glands before its transmission to the vertebrate host.     

Epidemiologic aspects of bovine anaplasmosis in semiarid range conditions of south central Idaho

The prevalance of Anaplasma marginale-infected cows, as determined by use of the modified rapid card agglutination (MRCA) test, was measured during a 4-year period (1980-1983). The prevalence of A marginale-infected cows, defined as positive reactors on the MRCA test, remained constant (31%-37%). The apparent incidence of A marginale transmission to susceptible cows was approximately 7% from 1980 to 1981, 8% from 1981 to 1982, and no transmission from 1982 to 1983. The occasional MRCA-positive cow became negative on the MRCA test, and 1 cow was determined to be free of A marginale infection by subinoculation of 100 ml of the cow's blood into a susceptible, splenectomized calf. Dermacentor andersoni, a known vector of A marginale, was often found on the cattle and in their environment. However, A marginale was not transmitted to susceptible, splenectomized calves, using collected ticks. Of 56 calves born to MRCA-positive cows, 82% were MRCA-positive within the first 3 months of life. These calves converted to MRCA-negative status and were determined to be free of A marginale infection by subinoculation of their blood into susceptible, splenectomized calves, indicating the passive transfer of colostral antibodies.     

Haemolysis in gel test for detecting bovine antibodies to Brucella abortus lipopolysaccharide

Optimal reagent parameters for a haemolysis in gel test for detection of bovine anti-Brucella abortus antibody were established. Using an alkali-treated crude lipopolysaccharide antigen attached to J-negative bovine erythrocytes, 0.125 mg of antigen, 0.1 ml of erythrocytes and 0.4 ml of guinea pig complement were required to perform 15 tests with appropriate serum controls. The haemolysis in gel test and an IgM radioimmunoassay (RIA) procedure were compared for their ability to detect the onset of increased serum antibody levels and antibody levels in sera diluted to extinction. While the RIA was more sensitive in amount of antibody detected, the haemolysis in gel test was equally able to detect initiation of antibody synthesis.