Artificial incubation of California condor Gymnogyps californianus eggs removed from the wild

The currrent California condor (Gymnogyps californianus) recovery plan entails increasing the reproductive rate via replacement-clutch manipulation of eggs. During the period from 1983 to 1985, 15 eggs were removed from wild nesting pairs for artificial incubation. The eggs were incubated at a dry bulb temperature of 36.4°C in modified forced-air Lyon Electric incubators. The incubation humidity was adjusted for individual eggs based on weight loss data (water = weight), 25.6–30.0°C wet bulb (41.0–63.0% Relative Humidity (RH)). The chicks were hatched initially under forced-air conditions of 36.1°C dry bulb, 31.1–01.7°C wet bulb (70.0–73.0% RH). In 1984, hatching parameters were changed to still-air conditions, 36.1°C dry bulb (top of the egg), 35.0°C dry bulb (bottom of the egg), 31.1–31.7°C wet bulb (70.0-73.0% RH). Tactile and auditory stimulation was utilized during the pip-to-hatch interval. From among 15 eggs collected, 13 hatched, and 12 condor chicks were raised successfully (hatchability: 86.7%; survivability: 92.3%).     

Heart sarcolemmal Ca2+ transport in endotoxin shock: I. Impairment of ATP-dependent Ca2+ transport

Effects of endotoxin administration on the ATP-dependent Ca²⁺ transport in canine cardiac sarcolemma were investigated. The results show that the sidedness of the sarcolemmal vesicles was not affected but the ATP-dependent Ca²⁺ transport in cardiac sarcolemma was decreased by 22 to 46% (p < 0.05) at 4 h following endotoxin administration. The kinetic analysis indicates that the V_max for ATP and for Ca²⁺ were decreased by 50% (p < 0.01) and 32% (p < 0.01), respectively, while the Km values for ATP and Ca²⁺ were not significantly affected after endotoxin administration. Magnesium (1–5 mM) stimulated while vanadate (0.25–3.0 M) inhibited the ATP-dependent Ca²⁺ transport, but the Mg²⁺-stimulated and the vanadate-inhibitable activities remained significantly lower in the endotoxin-treated animals. These data demonstrate that endotoxin administration impairs the ATP-dependent Ca²⁺ transport in canine cardiac sarcolemma and that the impairment is associated with a mechanism not affecting the affinity towards ATP and Ca²⁺. Additional experiments show that the Ca²⁺ sensitivity of the Ca²⁺-ATPase activity was indifferent between the control and endotoxic groups suggesting that endotoxic injury impairs Ca²⁺ pumping without affecting Ca²⁺-ATPase activity. Since sarcolemmal ATP-dependent Ca²⁺ transport plays an important role in the regulation of cytosolic Ca²⁺ homeostasis, an impairment in the sarcolemmal ATP-dependent Ca²⁺ transport induced by endotoxin administration may have a pathophysiological significance in contributing to the development of myocardial dysfunction in endotoxin shock.     

Formation of glycine conjugate and (-)-(R)-enantiomer from (+)-(S)-2-phenylpropionic acid suggesting the formation of the CoA thioester intermediate of (+)-(S)-enantiomer in dogs.

It has been proposed that the chiral inversion of the 2-arylpropionic acids is due to the stereospecific formation of the (-)-R-profenyl-CoA thioesters which are putative intermediates in the inversion. Accordingly, amino acid conjugation, for which the CoA thioesters are obligate intermediates, should be restricted to those optical forms which give rise to the (-)-R-profenyl-CoA, i.e., the racemates and the (-)-(R)-isomers. We have examined this problem in dogs with respect to 2-phenylpropionic acid(2-PPA). Regardless of the optical configuration of 2-phenylpropionic acid administered, the glycine conjugate was the major urinary metabolite and this was shown to be exclusively the (+)-(S)-enantiomer by chiral HPLC. Both (-)-(R)- and (+)-(S)-2-phenylpropionic acid were present in plasma after the administration of either antipode, and further evidence of the chiral inversion of both enantiomers was provided by the presence of some 25% of the opposite enantiomer in the free 2-phenylpropionic acid and its glucuronide excreted in urine after administration of (-)-(R)- and (+)-(S)-2-phenylpropionic acid. The (+)-(S)-enantiomer underwent chiral inversion to the (-)-(R)-antipode when incubated with dog hepatocytes. These data suggests that both enantiomers of 2-phenylpropionic acid are substrates for canine hepatic acyl CoA ligase(s) and thus undergo chiral inversion, but that the CoA thioester of only (+)-(S)-2-phenylpropionic acid is a substrate for the glycine N-acyl transferase. These studies are presently being extended to the structure and species specificity of the reverse inversion and amino acid conjugation of profen NSAIDs.     

Crystal structure of the Y52F/Y73F double mutant of phospholipase A2: increased hydrophobic interactions of the phenyl groups compensate for the disrupted hydrogen bonds of the tyrosines.

The enzyme phospholipase A2 (PLA2) catalyzes the hydrolysis of the sn-2 ester bond of membrane phospholipids. The highly conserved Tyr residues 52 and 73 in the enzyme form hydrogen bonds to the carboxylate group of the catalytic Asp-99. These hydrogen bonds were initially regarded as essential for the interfacial recognition and the stability of the overall catalytic network. The elimination of the hydrogen bonds involving the phenolic hydroxyl groups of the Tyr-52 and -73 by changing them to Phe lowered the stability but did not significantly affect the catalytic activity of the enzyme. The X-ray crystal structure of the double mutant Y52F/Y73F has been determined at 1.93 A resolution to study the effect of the mutation on the structure. The crystals are trigonal, space group P3(1)21, with cell parameters a = b = 46.3 A and c = 102.95 A. Intensity data were collected on a Siemens area detector, 8,024 reflections were unique with an R(sym) of 4.5% out of a total of 27,203. The structure was refined using all the unique reflections by XPLOR to a final R-factor of 18.6% for 955 protein atoms, 91 water molecules, and 1 calcium ion. The root mean square deviation for the alpha-carbon atoms between the double mutant and wild type was 0.56 A. The crystal structure revealed that four hydrogen bonds were lost in the catalytic network; three involving the tyrosines and one involving Pro-68. However, the hydrogen bonds of the catalytic triad, His-48, Asp-99, and the catalytic water, are retained. There is no additional solvent molecule at the active site to replace the missing hydroxyl groups; instead, the replacement of the phenolic OH groups by H atoms draws the Phe residues closer to the neighboring residues compared to wild type; Phe-52 moves toward His-48 and Asp-99 of the catalytic diad, and Phe-73 moves toward Met-8, both by about 0.5 A. The closing of the voids left by the OH groups increases the hydrophobic interactions compensating for the lost hydrogen bonds. The conservation of the triad hydrogen bonds and the stabilization of the active site by the increased hydrophobic interactions could explain why the double mutant has activity similar to wild type. The results indicate that the aspartyl carboxylate group of the catalytic triad can function alone without additional support from the hydrogen bonds of the two Tyr residues.     

Eumycetoma caused by Curvularia lunata in a dog

Curvularia lunata was cultured from black granules found in granulomatous tumefactions excised from the subcutis of a three year old Medium Schnauzer dog. Draining sinuses were present in some of the tumefactions. Accordingly the diagnosis of eumycotic mycetoma was made. This diagnosis was confirmed by histopathological examination. During the four years following the first surgical intervention, several more similar tumefactions were excised on three different occasions. The dog died of chronic renal failure at the age of 8 years. There was no bone involvement or visceral diffusion of the fungus. The granules were examined by scanning electron microscopy. Immunoglobulins in the dog's serum, assessed by a qualitative test, proved to be equal to immunoglobulins in the serum of a control dog. Precipitating antibodies against C. lunata were not found. The dog was treated for 150 days with itraconazole. In spite of good initial results, recurrence of the fungal lesions were observed after the treatment's interruption. Further treatment with itraconazole for 45 days proved ineffective. No side effects of the drug were observed. This is, to the best of our knowledge, the first case in which C. lunata is identified as the causative agent of an animal eumycetoma.     

Experiments with trap crops against African bollowrm, Heliothis armigera, in Ethiopia

In a 1 ha haricot bean (HB) (Phaseolus vulgaris L.) field strip-cropped with 2 rows of maize (MZ) (Zea mays L.) on either side and then at 25 m intervals, the number of African bollworm (ABW) (Heliothis armigera (Hubner) (Lepidoptera: Noctuidae)) eggs + larvae on MZ was higher than that on HB by a factor of nearly 18. In a replicated experiment where five trap crops were compared with HB, as a check, all of them contained significantly greater numbers of ABW eggs + larvae than the check. Among the crops tested, lupin (LP) (Lupinus termis Forsk.) and pigeon pea (Cajanus cajan (L.) Millsp.) followed by hyacinth bean (Dolichos lalab L.) contained the highest ABW egg + larval population while sunflower (Helianthus annuus L.) contained the lowest; MZ was intermediate. On the other hand, HB plots bordered with MZ suffered significantly lower percent pod damage by ABW than did the check. Although grain yield differences among treatments were not significant, MZ-bordered plots gave nearly 22% higher yield than the check during the heavy ABW infestation in 1982. These differences were less obvious during the 1983 season because of low ABW infestation. In hot pepper (Capsicum spp.) strip-cropped with lupin, egg + larval counts on the latter were greater by a factor of nearly four in 1981 and 18 in 1982. It is concluded that trap-cropping has a great potential in the integrated management of ABW.

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Résumé En 1981, dans un champ de haricot (Phaseolus vulgaris), des cultures pièges de maïs (Zea mays) ont été semées en rangs doubles tous les 25 m et de chaque côté du champ, soit un total de 5 rangs doubles. Le nombre d'H. armigera (oeufs + chenilles) a été près de 18 fois plus élevé sur maïs que sur haricot.En 1982 et 1983, l'attractivité de 5 cultures pièges: Dolichos lablab, Lupinus termis, Z. mays, Cajanus cajan et Helianthus annuus, a été comparée à celle du haricot. Toutes ces plantes portaient un nombre d'oeufs et de chenilles supérieur au témoin (P. vulgaris). L'ordre décroissant des contaminations était: L. termis, C. cajan, D. lablab, Z. mays et H. annuus. Les rangs de P. vulgaris bordés par des rangs de Z. mays étaient moins contaminés que les témoins. Bien que les différences de rendements en graines suivant les expériences n'aient pas été significatives, les parcelles bordées de maïs ont fourni une récolte supérieure de 22%, pendant l'année 1982, où la contamination par H. armigera a été forte. En 1983, où l'infestation a été faible, les différences étaient moins claires.En 1981 et 1982, 5 rangs de L. tremis à 25 m d'intervalle ont été utilisés comme pièges pour protéger le piment fort (Capsicum spp.). Les décomptes d'oeufs et de chenilles ont été respectivement près de 4 et 18 fois plus élevés.La conclusion est que les cultures pièges ont un grand avenir dans la lutte intégrée contre H. armigera.

     

The seasonality of phytoplankton in African lakes

Although some study of the subject began in 1899, wide-ranging information from African water-bodies has only become available since 1950. Important developments included the establishment of long-term centres of research, the adoption of improved methods for quantitative algal sampling, the more intensive study of environmental conditions, the beginnings of experimental testing, and the improvement of taxonomic knowledge.At higher latitudes (> 20 °) examples of pronounced algal seasonality are long-established; they are accompanied and influenced by marked changes in radiant energy income and so water temperature, and often by effects of seasonal water input. Illustrations are given from lakes in Morocco and South Africa.More generally in Africa, including the tropical belt, annual patterns of phytoplankton seasonality are usually either dominated by hydrological features (water input-output) or by hydrographic ones (water-column structure and circulation). Examples of both types are discussed, together with instances (e.g. L. Volta) of combined hydrological and hydrographic regulation. In both the seasonal abundance of diatoms is often distinct and complementary to that of blue-green algae, with differing relationships to vertical mixing and water retention.Horizontal variability in the seasonal cycle is especially pronounced in the larger or morphometrically subdivided lakes. Some inshore-offshore differentiation is also known to affect phytoplankton quantity (e.g. L. George) and species composition (e.g. L. Victoria). Longitudinal differentiation is common in elongate basins especially when with a massive or seasonal inflow at one end (e.g. L. Turkana, L. Nubia, L. Volta); occasional terminal upwelling can also be influential (e.g. southern L. Tanganyika). Such examples grade into the longitudinally differentiated seasonality of flowing river-reservoir systems, as studied on the Blue and White Niles.The annual amplitude of population density, expressed in orders of magnitude (=log₁₀ units), is one measure of seasonal variability. It can exceed 3 orders both in systems subject to hydrological wash-out (e.g. Nile reservoirs) and in the more variable species components of lakes of long retention (e.g. L. Victoria). Low amplitudes can be characteristic of some components (e.g. green algae in L. Victoria) or of total algal biomass (e.g. L. George, L. Sibaya).Seasonal changes may be subordinated to inter-annual ones, especially in shallow and hydrologically unstable lakes (e.g. L. Nakuru).     

Induction and release of secondary seed dormancy in genetically pure lines of Avena fatua

Induction and release of secondary dormancy in genetically pure dormant (AN-51, Mont 73) and non-dormant (CS-40, SH-430) lines of wild oat ( Avena fatua L.) were studied. These lines differed with regard to the optimal period of anaerobiosis necessary for induction of dormancy, and/or the degree (% of seeds acquiring dormancy) and duration of the dormancy induced. Secondary dormancy could be induced more effectively in the after-ripened seeds of dormant lines than in the non-dormant lines, where only a short-term dormancy could be induced (in 5–7 week-old-seeds). Higher anaerobiosis temperatures were more effective in inducing dormancy in all lines studied. Thus, as with primary dormancy, wild oat biotypes exhibit genetic variability in their secondary dormancy behaviour and factors like temperature can modify the expression of this trait.
The germination stimulants kinetin, isopentenyl adenine, sodium azide, potassium nitrate, ethanol and substituted phthalimides, which break primary dormancy in wild oats, stimulated germination of secondarily dormant seeds (line AN-51). Since these chemicals are structurally diverse, primary and secondary dormancies appear to be similar in part in their regulation.
Salicylhydroxamic acid, an inhibitor of cyanide-insensitive (alternative) respiration, did not inhibit: 1, spontaneous release of secondary dormancy in the line SH-430; and 2, stimulation of germination of secondarily dormant AN-51 seeds by various chemicals (except azide), suggesting that this respiratory pathway is not necessary for the release of induced dormancy.     

Trypanosoma brucei: analysis of relapsing populations in sensitive and resistant breeds of cattle

The clone DiTat 1.1 of Trypanosoma brucei brucei was injected into four bovids, and clones obtained from successive waves of parasitemia were used to study the expressed variant-specific surface glycoprotein repertoire. Twenty-four clones were obtained which could be classified into 12 different variable antigen types, in addition to the clone injected, using agglutination or immunofluorescence with monospecific antisera. The variable surface glycoproteins of the 25 clones were extracted using the detergent octyl-beta-D-glucopyranoside in the presence of the protease inhibitor, N-cbz-L-phenylalaninechloromethylketone. The molecular weights varied from 52,000 to 69,000 and the pI from 5.0 to 8.8. The virulence of 14 clones representing 13 variable antigen types was ascertained in mice. The mean survival time ranged from 20.5 to 43.0 days. Clones isolated from early peaks of parasitemia in the bovid were the most virulent while clones derived from later peaks were less virulent. It seems that organisms of diminishing virulence appear in bovids, leading to self-cure of the disease. All clones were sensitive to human serum in a blood infectivity inhibition test. Antibody against all virulent clones appeared in 20 cattle (10 Zebus, 10 Baoulés) which had been injected with T. brucei DiTat 1.1. There was no evidence for parasites of high or low virulence being preferentially expressed in resistant or sensitive hosts.