The development of Cowdria ruminantium in neutrophils

The sequential development of C. ruminantium (Kwanyanga and Kümm isolates) was followed in caprine leukocyte cultures by light microscopy, direct immunofluorescent microscopy (DFA), indirect immunoflourescent microscopy (IFA) and transmission electron microscopy (TEM). During the febrile response, one to several small cocci, large ring forms or rods were observed in neutrophils in blood smears and cytopreparations of neutrophil fractions using Diff Quik stain, Giemsa stain, DFA and TEM. One to several C. ruminantium colonies were seen in up to 35% of neutrophils maintained in vitro for 18 h to 5 days. The organisms were located in neutrophil phagosomes by TEM and were enveloped by two trilamellar unit membranes. Initially, C. ruminantium was tightly enclosed within phagosomes. At 20 h of incubation, organisms were frequently observed undergoing binary fission within enlarged phagosomal vacuoles. At later time periods, neutrophils harboured fully formed colonies (morula) containing numerous organisms. An occasional C. ruminantium-infected macrophage (Kümm isolate), and an occasional infected eosinophil (Kümm and Kwanyanga isolate) were found.     

In vitro induction of swine peripheral blood monocyte proliferation by the fibroblast-derived murine hematopoietic growth factor CSF-1

The addition of conditioned medium from murine L929 fibroblasts (MGF) to cultures of swine peripheral blood mononuclear cells (MNL) resulted in growth of cells of macrophage/monocyte lineage (MO). Glass-adherent swine MNL, shown to be greater than 95% phagocytic MO, grew in the presence of MGF, whereas swine blood granulocytes and lymphocytes were not MGF-responsive. Primary and secondary MO growth were directly dependent on MGF presence and concentration. MGF-stimulated MO synthesized DNA, as measured by cellular incorporation of tritium-labeled thymidine (3H-TdR). This mitogenic response was maximal by 5 to 6 days in primary MO cultures and declined thereafter to a lower magnitude in secondary MO cultures. In the presence of MGF, viable MO numbers increased with an approximate population doubling time of 5 to 7 days in primary culture. This growth rate was prolonged, to about 10 to 12 days, for MGF-stimulated MO in secondary cultures. MGF removal from primary and secondary MO cultures resulted in rapid growth cessation and cell death. MGF-stimulated MO could not be sustained in secondary culture beyond 7 weeks. MGF-cultured MO were positive for latex phagocytosis, non-specific esterase, Fc-receptor expression, and could mediate antibody-dependent cell-mediated cytotoxicity. The MO-mitogenic principle of MGF was identified as the murine, macrophage-specific colony-stimulating factor, CSF-1 (M-CSF). The swine MO-proliferative response to MGF was inhibited by addition of monospecific goat antisera to M-CSF. Purified M-CSF stimulated the growth of swine MO from cultures of MNL and primary glass-adherent MO.     

Ovarian activity in F1 prepubertal ewe lambs under tropical conditions

Seventy crossbred ewe lambs, born from Pelibuey (PB) dams mated with five sire breeds: White Dorper × PB (WD × PB), Black Head Dorper × PB (BHD × PB), Ile de France × PB (Ile × PB) Katahdin × PB (KT × PB) and Pelibuey (PB × PB), were used to determine age and weight to puberty, follicular populations during prepubertal development, ovulation rate at first ovulation and function of the first corpus luteum (FCL). Ewe lambs were maintained under grazing conditions and were supplemented with 300 g head^− 1 day^− 1 of a concentrate with 14% CP and 3.0 Mcal ME kg^− 1 of DM. From 145 days of age, monthly observations were performed by endoscopy to monitor follicular populations and luteal structures. Ovarian follicles were classified according to size into small (≥ 1 and < 3 mm), medium (≥ 3 and ≤ 4 mm) and large (> 4 mm). Additionally, weekly observations by ultrasonography were performed in four ewes per genotype to assess follicular populations. Luteal function from first ovulation was determined by circulating concentrations of progesterone. Ovulation rate was not affected by genotype (P > 0.05). All breed groups had the same pattern of follicular populations, where most of follicles were small, followed by medium and a small proportion of large follicles. Maximum diameter of follicles was similar for all genotypes during prepubertal development (P > 0.05), with an average range of 3.9 ± 0.20 to 4.4 ± 0.20 mm. The BHD × PB reached puberty at the youngest age of 240.9 ± 13.0 days, compared to all other genotypes (P < 0.05), 259.1 ± 11.7 for WD × PB, 279.3 ± 9.6 for KT × PB and 289.0 ± 15.3 days for PB × PB and 308.6 ± 11.9 days for Ile × PB ewe lambs. Ile × PB had greater weight to first ovulation (31.5 ± 1.19 kg) as compared to other breed groups (P < 0.01). The largest percentage of FCL was for KT × PB ewes (100%) (P < 0.05), followed by BHD × PB (88.9%), PB × PB (83.3), WD × PB (78.6%) and Ile × PB (77.8%). There were no differences in ovulation rate attributable to type of lambing from which ewe lambs came from (P > 0.05). BHD × PB ewe lambs reached puberty at an earlier age than other in this study. KT × PB ewe lambs had the highest percentage of FCL in their first ovulation. Breed of sire Ile de France increased weight to puberty and decreased FCL at first ovulation. In conclusion, the BHD × PB ewe showed puberty at an earlier age than other genotypes, while the KT × PB had the highest percentage of functional corpus luteum in their first ovulation. Ewe lambs coming from single lambings reach puberty at a heavier weight than those coming from multiple lambings. The ovulation rate in the first ovulation of ewe lambs is not affected by genotype and type of lambing.     

Survival of Post‐larval Litopenaeus vannamei Following Acclimation to Low Salinity Waters at Different Temperatures

Temperature and salinity are two factors known to influence the growth potential and survival of the Pacific white shrimp, Litopenaeus vannamei, acclimated to low salinity waters. In west Alabama, farmers suspect low water temperatures at stocking, in conjunction with low salinity and suboptimal ionic profiles, might be responsible for reduced survival and production at harvest. To determine the influence of temperature and salinity on post‐larval (PL) L. vannamei, a series of bioassays were conducted at the E.W. Shell Fisheries Research Station in Auburn, Alabama and Claude Peteet Mariculture Center in Gulf Shores, Alabama. PL L. vannamei of ages 11, 13, and 20 (PL₁₁, PL₁₃, and PL₂₀) were acclimated down to salinities of 12, 4, 2, 1, 0.5, and 0.2 ppt at different temperatures ranging from 17.6 to 24.0 C. During the acclimation bioassays survivals were assessed at 24 and 48 h. PL survival of the three age groups examined were significantly reduced at salinities of 1, 0.5, and 0.2 ppt. These results correspond well to those reported at higher temperatures confirming that across the tested temperature range salinity endpoint was the driving factor in determining survival and that suboptimal temperatures had a minimal influence on survival.     

ADVERSE EFFECTS OF HUMIC SUBSTANCES FROM DIFFERENT ORIGIN ON LUPIN AS RELATED TO IRON SOURCES

Humic substances improve the efficiency of different iron (Fe) sources overcoming Fe deficiency chlorosis of plants. However, applied at high rates, they can promote negative effects on plants. The main objective of this work was to study the potential adverse effect of three humic acids from different origin when they were applied with two effective Fe sources for plants: Fe- ethylenediaminedihydroxyphenylacetic acid (EDDHA) and Vivianite. To this end, an experiment with lupin (Lupinus albus L.) was performed involving two factors: (i) Fe source, and (ii) humic substances from three different origin (composted cork, leonardite, and compost obtained from a mixture of olive husk with cotton gin trash) applied at 0, 0.1, and 0.5 g organic carbon (C) kg^?1 of growing media. At the rates used, humic substances promoted adverse effects on plant development, chlorophyll meter readings, and Fe content in lupin grown in calcareous media. Overall, the effect on dry matter and Fe content in plants was more relevant when Fe was supplied with Vivianite, the effect on chlorophyll meter readings being more significant when Fe was applied as Fe-EDDHA. Differences were also observed depending on the source of humic substances, those from leonardite promoting the greatest decrease in dry matter in roots and shoots. These humic substances possessed the highest values of spectroscopy index for aromaticity (A₂₅₄ ). On the other hand, the application of humic substances from olive husk compost, which exhibited the lower aromaticity index, resulted in the smallest decrease in dry matter production and chlorophyll meter readings. Dry matter in roots decreased logarithmically with increased values of the estimates of the amounts of aromatic compounds accumulated in the growing media (R² = 0.92; P < 0.01) with Vivianite as Fe source. Thus, the effects decreasing dry matter production, particularly in roots, and chlorophyll meter readings can be ascribed at least partially to the presence of phytotoxic aromatic compounds in humic substances.     

Extraction of viral DNA from erythrocytes of swine with acute African swine fever

The preparation of wild-type African swine fever (ASF) virus DNA from small amounts of viremic blood from acutely febrile pigs is outlined. The extracted DNA is viral and not host-cell DNA, because of specific homology with cell culture grown and purified ASF virus and because no DNA bands are obtained with an equal amount of nonviremic pig blood. Thus, in the absence of suitable serologic methods for strain identification, it is now possible to catalogue wild-type isolates by characteristic DNA restriction patterns. The wild-type virus genome contains terminal single-stranded DNA cross-links and has the largest genome size (180 kilobase pairs) reported for the ASF virus. Experimental passage of the virus in contact-infected pigs and buffy coat cultures appears to confirm the stable nature of the ASF genome in the field.     

Physicochemical Characterization of Nopal Pads (Opuntia ficus indica) and Dry Vacuum Nopal Powders as a Function of the Maturation

This paper presents the physicochemical and nutrimental characterization of fresh nopal (Opuntia ficus indica, Redonda variety) and nopal powder produced at different stages of development. Nopal powder was obtained by dry vacuum technique using 10² Torr and low temperature (40°C). The results showed that the nutrimental and mineral composition of nopal changes as a function of the maturation as follow: The ash content increases from 18.41 for nopalitos (60 g of weight) to 23.24% (nopal pads 200 g); calcium content increases from 1.52 to 3.72%, while phosphorous exhibits an opposite trend: 0.43 to 0.27%, respectively. Calcium oxalate was determined by X-ray diffraction and SEM microscopy and quantified by using atomic absorption spectroscopy. Calcium oxalate decreases from 7.95 to 3.47 mg/g and the Ca/P ratio varies from 3.6 to 11. The soluble fibre decreases from 25.22 to 14.91%, while insoluble fibre increases from 29.87 to 41.65%. These results suggest that nopal could be an important source of minerals within the diets of people in Mexico and the rest of Latin America.     

For services rendered? Modeling hydrology and livelihoods in Andean payments for environmental services schemes

In the Andes, demand for water is growing and upland land-use changes are increasing. Water quality, quantity and seasonal flow have thus also become environmental services with potential monetary value. Yet, currently the region's pioneer PES schemes are not paying for measured environmental services, but for proxy land uses thought to provide the(se) service(s). Hydrological modeling makes explicit the tacit causal relationships and tests underlying assumptions. Ideally, when combined with an economic analysis of land-use alternatives, this could inform decision makers on how much to pay for different interventions in different spatial locations. This paper focuses on two Andean watersheds: Moyobamba (Peru) and Pimampiro (Ecuador). In the first case, a municipal water company is preparing a payment for environmental services (PES) scheme to reduce upstream sediment loads. In the second, a similar conservation-oriented municipal PES scheme has operated since 2000, but the hydrological linkages have never been tested. Applying the Soil & Water Assessment Tool (SWAT), we identify in both watersheds biophysically critical areas for service delivery, and compare services for current land uses with change scenarios: deforestation, reforestation, live barriers, and agroforestry. We then use the ECOSAUT optimization model to predict net economic benefits for service providers. In Moyobamba, switching to shade-grown coffee would halve sediment yields, and increase significantly farmers’ economic benefits. This requires high up-front investment, but the willingness to pay of water users in Moyobamba town may suffice to cover the upfront costs. In Pimampiro, resumed deforestation would increase sediments by >50% and reduce dry-season flow by 0.5%, thus reinforcing the rationale of the existing PES scheme, focused on conserving native forests and grasslands.