Multistep Solid-State Organic Synthesis of Carbamate-Linked Covalent Organic Frameworks.

Herein, we demonstrate the first example of a multistep solid-state organic synthesis, in which a new imine-linked two-dimensional covalent organic framework (COF-170, 1) was transformed through three consecutive postsynthetic modifications into porous, crystalline cyclic carbamate and thiocarbamate-linked frameworks. These linkages are previously unreported and inaccessible through de novo synthesis. While not altering the overall connectivity of the framework, these chemical transformations induce significant conformational and structural changes at each step, highlighting the key importance of noncovalent interactions and conformational flexibility to COF crystallinity and porosity. These transformations were assessed using 15N multiCP-MAS NMR spectroscopy, providing the first quantitation of yields in COF postsynthetic modification reactions, as well as of amine defect sites in imine-linked COFs. This multistep COF linkage postsynthetic modification represents a significant step toward bringing the precision of organic solution-phase synthesis to extended solid-state compounds.

Conversion of Imine to Oxazole and Thiazole Linkages in Covalent Organic Frameworks.

Imine-linked ILCOF-1 based on 1,4-phenylenediamine and 1,3,6,8-tetrakis(4-formylphenyl)pyrene was converted through consecutive linker substitution and oxidative cyclization to two isostructural covalent organic frameworks (COFs), having thiazole and oxazole linkages. The completeness of the conversion was assessed by infrared and solid-state NMR spectroscopies, and the crystallinity of the COFs was confirmed by powder X-ray diffraction. Furthermore, the azole-linked COFs remain porous, as shown by nitrogen sorption experiments. The materials derived in this way demonstrate increased chemical stability, relative to the imine-linked starting material. This constitutes a facile method for accessing COFs and linkages that are otherwise difficult to crystallize due to their inherently limited microscopic reversibility.

Chemical Conversion of Linkages in Covalent Organic Frameworks.

The imine linkages of two layered, porous covalent organic frameworks (COFs), TPB-TP-COF ([C6H3(C6H4N)3]2[C6H4(CH)2]3, 1) and 4PE-1P-COF ([C2(C6H4N)4][C6H4(CH)2]2, 2), have been transformed into amide linkages to make the respective isostructural amide COFs 1' and 2' by direct oxidation with retention of crystallinity and permanent porosity. Remarkably, the oxidation of both imine COFs is complete, as assessed by FT-IR and 13C CP-MAS NMR spectroscopy and demonstrates (a) the first chemical conversion of a COF linkage and (b) how the usual "crystallization problem" encountered in COF chemistry can be bypassed to access COFs, such as these amides, that are typically thought to be difficult to obtain by the usual de novo methods. The amide COFs show improved chemical stability relative to their imine progenitors.

Chemistry of Covalent Organic Frameworks.

Linking organic molecules by covalent bonds into extended solids typically generates amorphous, disordered materials. The ability to develop strategies for obtaining crystals of such solids is of interest because it opens the way for precise control of the geometry and functionality of the extended structure, and the stereochemical orientation of its constituents. Covalent organic frameworks (COFs) are a new class of porous covalent organic structures whose backbone is composed entirely of light elements (B, C, N, O, Si) that represent a successful demonstration of how crystalline materials of covalent solids can be achieved. COFs are made by combination of organic building units covalently linked into extended structures to make crystalline materials. The attainment of crystals is done by several techniques in which a balance is struck between the thermodynamic reversibility of the linking reactions and their kinetics. This success has led to the expansion of COF materials to include organic units linked by these strong covalent bonds: B-O, C-N, B-N, and B-O-Si. Since the organic constituents of COFs, when linked, do not undergo significant change in their overall geometry, it has been possible to predict the structures of the resulting COFs, and this advantage has facilitated their characterization using powder X-ray diffraction (PXRD) techniques. It has also allowed for the synthesis of COF structures by design and for their formation with the desired composition, pore size, and aperture. In practice, the modeled PXRD pattern for a given expected COF is compared with the experimental one, and depending on the quality of the match, this is used as a starting point for solving and then refining the crystal structure of the target COF. These characteristics make COFs an attractive class of new porous materials. Accordingly, they have been used as gas storage materials for energy applications, solid supports for catalysis, and optoelectronic devices. A large and growing library of linkers amenable to the synthesis of COFs is now available, and new COFs and topologies made by reticular synthesis are being reported. Much research is also directed toward the development of new methods of linking organic building units to generate other crystalline COFs. These efforts promise not only new COF chemistry and materials, but also the chance to extend the precision of molecular covalent chemistry to extended solids.

Local Electronic Structure of Molecular Heterojunctions in a Single-Layer 2D Covalent Organic Framework.

The synthesis of a single-layer covalent organic framework (COF) with spatially modulated internal potentials provides new opportunities for manipulating the electronic structure of molecularly defined materials. Here, the fabrication and electronic characterization of COF-420: a single-layer porphyrin-based square-lattice COF containing a periodic array of oriented, type II electronic heterojunctions is reported. In contrast to previous donor-acceptor COFs, COF-420 is constructed from building blocks that yield identical cores upon reticulation, but that are bridged by electrically asymmetric linkers supporting oriented electronic dipoles. Scanning tunneling spectroscopy reveals staggered gap (type II) band alignment between adjacent molecular cores in COF-420, in agreement with first-principles calculations. Hirshfeld charge analysis indicates that dipole fields from oriented imine linkages within COF-420 are the main cause of the staggered electronic structure in this square grid of atomically-precise heterojunctions.

Palladium-catalysed anti-Markovnikov selective oxidative amination.

In recent years, the synthesis of amines and other nitrogen-containing motifs has been a major area of research in organic chemistry because they are widely represented in biologically active molecules. Current strategies rely on a multistep approach and require one reactant to be activated prior to the carbon-nitrogen bond formation. This leads to a reaction inefficiency and functional group intolerance. As such, a general approach to the synthesis of nitrogen-containing compounds from readily available and benign starting materials is highly desirable. Here we present a palladium-catalysed oxidative amination reaction in which the addition of the nitrogen occurs at the less-substituted carbon of a double bond, in what is known as anti-Markovnikov selectivity. Alkenes are shown to react with imides in the presence of a palladate catalyst to generate the terminal imide through trans-aminopalladation. Subsequently, olefin isomerization occurs to afford the thermodynamically favoured products. Both the scope of the transformation and mechanistic investigations are reported.

Practical water production from desert air.

Energy-efficient production of water from desert air has not been developed. A proof-of-concept device for harvesting water at low relative humidity was reported; however, it used external cooling and was not desert-tested. We report a laboratory-to-desert experiment where a prototype using up to 1.2 kg of metal-organic framework (MOF)-801 was tested in the laboratory and later in the desert of Arizona, USA. It produced 100 g of water per kilogram of MOF-801 per day-and-night cycle, using only natural cooling and ambient sunlight as a source of energy. We also report an aluminum-based MOF-303, which delivers more than twice the amount of water. The desert experiment uncovered key parameters pertaining to the energy, material, and air requirements for efficient production of water from desert air, even at a subzero dew point.

Epidemiology and seasonal dynamics of gastrointestinal nematode infections of sheep in a semi-arid region of eastern Ethiopia.

A study on the epidemiology and seasonal dynamics of gastrointestinal nematode infections of sheep was carried out in a semi-arid region of eastern Ethiopia for 2.5 years (May 2003 to September 2005). The experimental flock comprised a total of 60 Black Head Ogaden sheep, consisting of four equal groups of young male and female and old male and female sheep. These grazed on communal pastures together with a larger university flock, as well as with animals owned by neighbouring small-holder farmers. A new experimental flock was established each year of the study. Parasitological data (EPG, faecal culture L3, PCV and FAMACHA estimates) and animal performance (weight change) were recorded each month on all experimental animals. In addition, four tracer lambs were assigned each month to the flock to determine the seasonal patterns of infective larvae acquired from pasture. Results showed distinct seasonal patterns associated with the bi-modal annual rainfall. High levels of infection occurred during the short and long rain seasons with peaks occurring in May and September of each year. Haemonchus contortus was the most prevalent parasite, followed by Trichostrongylus spp., with a number of other nematode species being occasionally recorded. H. contortus showed and increased propensity to undergo arrested development during the dry seasons. Correlations between EPG and PCV, EPG and FAMACHA eye scores, and PCV and FAMACHA eye scores were all highly significant (P<0.001). However, there was no significant association between the EPG and LW of the study animals during each study year. This information will provide a basis for developing epidemiologically based control strategies for gastrointestinal nematode parasites that are appropriate for flocks owned by small-holder farmers of semi-arid areas of eastern Ethiopia.

Prolonged persistence of fecally excreted ivermectin from reindeer in a sub-arctic environment.

In December 2001 and 2002, feces from reindeer calves treated with ivermectin were distributed on plots established on two types of forested reindeer pasture in northern Finland. The ungrazed plots were on an enclosure that had been fenced to prevent reindeer access for the last 6 years. The grazed plots were on an area that had been heavily stocked by reindeer during the last 5 years. After enclosures had been established, reindeer and large wildlife were prevented from entering by a fence. Topsoil samples (feces, vegetation, and soil) were collected monthly during the summers of the following 2 years, over a period of from 25 to 95 weeks after deposition. The samples were analyzed for ivermectin using HPLC. Although ivermectin degradation rapidly took place during the first spring, considerable residual ivermectin could be measured throughout the sampling time, showing that ivermectin in feces on pasture may not be photodegraded as rapidly as previously believed. The results support the need for further environmental evaluation studies on the use of ivermectin to control reindeer parasites.

Towards the implementation of the "basket of options" approach to helminth parasite control of livestock: emphasis on the tropics/subtropics.

The virtual reliance on anthelmintic drugs alone to control internal parasites of livestock is inappropriate and ultimately unsustainable. In the tropics and subtropics, widespread and high levels of anthelmintic resistance, particularly in nematode parasites of small ruminants, is rife. But more to the point, many farmers in these regions of the world are resource poor and cannot afford, or are reluctant to purchase drugs that may also be of dubious quality. As it is with any intervention, the benefits must outweigh the costs. This is not only in terms of conventional parameters such as reduced mortality and increasing productivity (meat, milk, fibre and traction power) of livestock, but also within the broad framework of helminths of veterinary/human importance, the aim should be a positive impact on reducing the threat of helminth zoonoses. However, understanding the issues involved and education of the end-users (farmers) is of fundamental importance, before any internal parasite control program should be promoted. Within the above context, we provide examples of how the "basket of options" approach could be adopted for the control of three quite disparate helminth problems in the tropics and subtropics, viz.: strongyle nematode infections of donkeys, the Taenia solium cysticercosis/taeniosis problem of pig and man and Haemonchus contortus infections in small ruminants. The "best practice" approaches can be defined as those "basket of options" that are practical, affordable, available and appropriate, whether to the commercial producer, or to the resource-poor farmer. Constraints that may restrict applying such options are accessibility to, and affordability of, suitable remedies and above all, the availability of information needed to make informed decisions in this regard.

Anthelmintic resistance of nematode parasites of small ruminants in eastern Ethiopia: exploitation of refugia to restore anthelmintic efficacy.

Faecal egg count reduction tests (FECRT) were conducted in May 2003 to determine the efficacy of anthelmintics used for treatment against nematode parasites in separately managed sheep and goat flocks at Alemaya University in eastern Ethiopia. These tests revealed high levels of anthelmintic resistance to albendazole, tetramisole, the combination of these two drugs, and to ivermectin in the goat flock (predominantly infected by Haemonchus contortus and Trichostrongylus spp.), whereas all drugs were highly efficacious in the sheep flock. A second FECRT confirmed these observations. Following this, a new management system was implemented on the goat flock for a period of 9 months (January-September 2004) in an attempt to restore the anthelmintic efficacy. This involved a combination of measures: eliminating the existing parasite infections in the goats, exclusion from the traditional goat pastures, and introducing communal grazing of the goats with the university sheep flock and livestock owned by neighbouring small-holder farmers. A second series of FECRTs (Tests 3 and 4) conducted 7 months after this change in management, showed high levels of efficacy to all three drugs (albendazole, tetramisole and ivermectin) in the goat flock. This is the first field study to demonstrate that anthelmintic efficacy in the control of nematode parasites of small ruminants can be restored by exploiting refugia.

Ethnoveterinary plant preparations as livestock dewormers: practices, popular beliefs, pitfalls and prospects for the future.

Ethnomedicine is an integral part of traditional medical practices in many countries of the developing world. A large proportion of the population uses this form of treatment for primary health care and for the treatment of ailments in their livestock. Livestock is a major asset for resource-poor smallholder farmers and pastoralists throughout the world and internal parasites are recognized by these communities as having an impact on livestock health. Parasitic infections are among those infections that traditional healers confidently treat and against which an enormous variety of remedies exist. Many of these are based on the use of plant preparations. Although various methods have been used for the validation of traditional phytomedical preparations, there is a lack of standardization of these procedures. The present study is aimed at providing an overview of ethnoveterinary deworming preparations, the various methods that have been used in their validation and the future prospects for their use against helminth parasites of ruminant livestock in developing countries, with an emphasis on nematode parasites. Recommendations are made on the procedures that should be followed to conduct in vivo and in vitro assays. Fostering better interaction between traditional healers and scientists is advocated to prevent harmful overexploitation, both of local knowledge and of plant species that may have effects against nematode parasites.

Nematode control in 'green' ruminant production systems.

Collectively, nematode parasites of domestic ruminants continue to pose the greatest disease problem in grazing livestock systems worldwide, despite the powerful and extensive chemotherapeutic arsenal available for their control. The widespread development of anthelmintic resistance, particularly in nematode parasites of small ruminants, and the trend towards non-chemical (ecological, organic, green) farming of livestock has provided an impetus for the research and development of alternative parasite control methods. This article provides a brief overview of the non-chemotherapeutic options for parasite control and how they might play a role either in organic farming or in other low-input farming systems.

Global perspectives on nematode parasite control in ruminant livestock: the need to adopt alternatives to chemotherapy, with emphasis on biological control.

Effective, sustainable control of nematode parasites of grazing livestock is becoming evermore challenging and difficult. This is largely due to two contrasting issues. One is the rapid escalation of resistance to anthelmintic drugs, which is arguably the greatest problem now facing the small ruminant industries worldwide. Secondly, there is the increasing trend towards organic farming, in which there is prohibition of the prophylactic use of all chemical compounds. Livestock producers urgently need non-chemotherapeutic alternatives in parasite control. Researchers have responded to this challenge and a variety of quite different approaches have been the subject of intense investigation in many countries for several decades now. These vary in relation to their stage of development for on-farm use, their utility, and their applicability across the spectrum of grazing livestock enterprises throughout the world. One relatively recent innovation is the biological control approach to nematode parasites. This has now reached the stage of commercialization. This review focuses on these issues and provides an overview of the possible ways in which the biological control of nematode parasites could be employed in grazing ruminant livestock systems worldwide.

Seasonal translation of infective larvae of gastrointestinal nematodes of cattle and the effect of Duddingtonia flagrans: a 3-year pilot study.

A study was conducted over 3 years (1998-2000) to investigate larval availability of gastrointestinal nematodes from faeces of cattle reared under different parasite control schemes. These cattle were part of a parallel, but separate grazing trial, and were used as donor animals for the faecal material used in this experiment. At monthly intervals, faeces were collected and pooled from three groups of first-season grazing cattle. These groups were either untreated, ivermectin bolus treated or fed the nematophagous fungus Duddingtonia flagrans. The untreated and fungus treated animals were infected with gastrointestinal nematodes and the number of eggs per gram (epg) pooled faeces ranged between 50 and 700 in the untreated group and between 25 and 525 epg in the fungus treated group. Each year between June and September, artificial 1 kg dung pats were prepared and deposited on pasture and protected from birds. The same treatments, deposition times and locations were repeated throughout the study. Larval recovery from herbage of an entire circular area surrounding the dung pats was made in a sequential fashion. This was achieved by clipping samples in replicate 1/4 sectors around the dung pats 4, 6, 8 and 10 weeks after deposition. In addition, coinciding with the usual time of livestock turn-out in early May of the following year, grass samples were taken from a circular area centred where the dung pats had been located to estimate the number of overwintered larvae, which had not been harvested during the intensive grass sampling the previous year. It was found that recovery and number of infective larvae varied considerably within and between seasons. Although the faecal egg counts in 1999 never exceeded 300 epg of the faecal pats derived from the untreated animals, the abnormally dry conditions of this year generated the highest level of overwintered larvae found on herbage in early May 2000, for the 3 years of the study. Overall, biological control with D. flagrans significantly reduced larval availability on herbage, both during and between the grazing seasons, when compared with the untreated control. However, the fungus did not significantly reduce overwintered larvae derived from early season depositions (June and July), particularly when dung pats disappeared within 2 weeks after deposition. Very low number of larvae (<3 per kg dry herbage) were sporadically recovered from grass samples surrounding the ivermectin bolus faecal pats.

Dung beetle activity and the development of trichostrongylid eggs into infective larvae in cattle faeces.

In the perspective to reduce the use of antiparasitic drugs, any interaction between the processes that control the development of parasite eggs into infective larval stages (L3) in dung and the activities of different dung-breeding organisms becomes of interest. The objective of the present study was to determine whether dung beetle activity affected the development of parasite larvae in cattle dung. Faeces containing eggs of parasites (predominantly Cooperia spp.) were pooled according to high (250-600 epg) or low (approx. 100 epg) egg counts. Experimental dung pats were formed for each category of faeces and to half of these pats, dung beetles (20 Aphodius rufipes, 20 A. scybalarius syn. rufus) were added and kept for 12 days at 21 degrees C (+/-1 degrees C) and 90% RH (+/-5% RH). Beetles were then removed and the pats were divided in two where half the pat was incubated for an additional 12 days at 21 degrees C (+/-1 degrees C) and 90% RH (+/-5% RH) and the other half was immediately analysed. A greater number of L3 were recovered from the dung subjected to beetle activity compared with control dung (P<0.001). However, following an additional 12 days incubation of the dung, similar numbers of L3 were recovered from beetle-affected dung with high egg counts, whereas there were significantly greater numbers of L3 derived from the control dung (P<0.001). There was no significant difference in L3 recovery in the two categories of pats (i.e. high and low nematode egg counts). The results indicate that activity by Aphodius spp. in fresh dung can optimise conditions for nematode development to the infective larval stage if favourable environmental conditions prevail. Such synergistic effect may be due to the fact that dung beetles used in this study are dung dwellers, i.e. no substantial amount of the dung is removed or burrowed as these beetles feed, lay their eggs and the larval development takes place in brood chambers inside the dung.

The anthelmintic efficacy of the plant, Albizia anthelmintica, against the nematode parasites Haemonchus contortus of sheep and Heligmosomoides polygyrus of mice.

Albizia anthelmintica Brong., belongs to the plant family Mimosaceae. The plant is widely used in East Africa by poor smallholder farmers and pastoralists to treat their livestock against internal parasites. The anthelmintic effects of water extracts from the bark of A. anthelmintica, obtained from three different geographic areas in Kenya and using different methods of preparation, were tested at different doses in sheep and mice infected with the nematode parasites Haemonchus contortus and Heligmosomoides polygyrus, respectively. Lambs were infected with 3000 infective larvae of H. contortus and treated with the plant preparations 28 days later, while mice were infected with 200 infective larvae of H. polygyrus and treated 18 days later. Proximate analysis established high levels of crude proteins in A. anthelmintica bark. Two sheep out of the 45 treated with the plant preparations suffered from transient bloat, which was relieved by dosing with a surfactant. Significant reductions in faecal egg counts were observed in lambs treated with A. anthelmintica in two of the three experiments undertaken, but the efficacy levels achieved were well below the 70% reduction required. Similar values of packed red cell volume and live weight gain were observed for treated and control lambs. There was no overall significant effect of treatment with A. anthelmintica on faecal egg and total worm counts in mice. A dose rate of 1000 mg/kg bodyweight of A. anthelmintica preparation resulted in death of all mice. The results show that A. anthelmintica at the doses and preparations used is not efficacious against H. contortus in sheep or against H. polygyrus in mice.

Evaluation of anthelmintic properties of extracts from some plants used as livestock dewormers by pastoralist and smallholder farmers in Kenya against Heligmosomoides polygyrus infections in mice.

Parasitic nematodes are among the most common and economically important infectious diseases of grazing livestock, especially in small ruminants in the tropics and subtropics in Kenya the control of gastrointestinal nematode infections in sheep and goats is usually made with synthetic anthelmintics but substantial levels of anthelmintic resistance have been recorded. A number of medicinal plants, that may provide possible alternatives, and are used by pastoralists and smallholder farmers in Kenya as deworming agents for their livestock and equines, namely Aframomum sanguineum, Dodonea angustifolia, Hildebrandtia sepalosa, Myrsine africana, Rapanea melanophloeos from Kenya, and Azadirachta indica from Kenya and Malaysia, together with the chemicals embelin and santonin that occur in some of these plants, were evaluated against Heligmosomoides polygyrus in mice. Commercial anthelmintics, namely ivermectin, pyrantel and piperazine, were also investigated, both to validate the mouse model system and to assess efficacy of these drugs against H. polygyrus. Pyrantel and ivermectin were highly effective in reducing the numbers of H. polygyrus worms as well as eggs in faeces of the mice, but piperazine had a lower activity. Application of santonin and M. africana significantly reduced the number of total worm counts (TWC) but not faecal egg counts (FEC). The use of embelin, R. melanophloeos and A. indica reduced FEC but not TWC. In all cases, however, reductions were well below the a priori level of 70% required for biological significance. A. sanguineum, D. angustifolia and H. sepalosa had no effect on either TWC or FEC. In conclusion, none of the plant preparations had any biologically significant anthelmintic effect in this monogastric host-parasite model system.

Evaluation of gastro-intestinal nematode parasite control strategies for first-season grazing cattle in Sweden.

A three-year grazing experiment (1998-2000) was conducted with first-season grazing cattle (FSGC) on improved pastures in central-eastern Sweden. Comparison was made between five groups with 10 calves in each group where four of these were set stocked and either (1) untreated, (2) ivermectin bolus treated, (3) subjected to biological control with the nematophagous fungus Duddingtonia flagrans, or (4) treated with a copper wire particle bolus. The fifth treatment was an evasive grazing strategy, whereby untreated calves were turned out onto pasture used by older cattle the previous year and then these calves were moved to silage aftermath in mid-July. To introduce low-levels of parasite infection to the experiment, each animal received a 'priming dose' of approximately 5,000 Ostertagia ostertagi and 5,000 Cooperia oncophora infective third stage larvae immediately prior to the start of the first grazing year of the trial. Results showed that efficient and sustainable parasite control of FSGC was possible to achieve without the use of anthelmintics by using turnout pastures that the previous year had been grazed by older cattle, in combination with a mid-July move to aftermath leys. Biological control also proved beneficial but the efficacy was impaired if high faecal egg counts coincided with rapid dung pat degradation due to heavy rainfall. No indication of parasite control was observed with the copper wire particle bolus. It was also demonstrated that the impact of gastrointestinal (GI) parasitism varied between years and that the level of overwintering contamination is important but likewise, is unpredictable. Although faecal egg counts in 1999 were low, due both to a delayed turnout and drought for the major part of the grazing season, deposited eggs successfully developed to infective larvae and overwintered in large numbers. The population of overwintered infective larvae at the time of turnout in early May played an important role in the course of infection in 2000 and resulted in an average 65 kg advantage of the ivermectin treated calves compared with the untreated calves.Thus, this three-year grazing experiment has emphasised the importance of subclinical gastrointestinal nematode infections in FSGC in Sweden. In addition, the study has shown that adequate parasite control may be achievable without the use of anthelmintics.

Anthelmintic activity of preparations derived from Myrsine africana and Rapanea melanophloeos against the nematode parasite, Haemonchus contortus, of sheep.

Myrsine africana L. and Rapanea melanophloeos L. belong to the plant family Myrsinaceae. Various rural communities in Kenya, such as smallholder farmers and pastoralists, use them to treat their livestock. The anthelmintic effects/activities of leaves and fruits of M. africana and fruits of R. melanophloeos were tested in sheep experimentally infected with the nematode parasite Haemonchus contortus. Male lambs were infected with 3000-5000 third stage larvae of H. contortus and treated 28 days after inoculation with concoctions made from leaves or fruits of the plants. No significant reduction in faecal nematode egg counts was observed with any of the concoctions at any of the doses tested. Packed red cell volume decreased and live weight increased at similar rates in treated and control groups, thus there was no significant effect of treatment. The results showed that the tested extracts of the M. africana and R. melanophloeos were not efficacious against H. contortus in sheep.