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As research funding becomes more competitive, it will be imperative for researchers to break the mentality of a single laboratory/single research focus and develop an interdisciplinary research team aimed at addressing real world challenges. Members of this team may be at the same institution, may be found regionally, or may be international. However, all must share the same passion for a topic that is bigger than any individual's research focus. Moreover, special consideration should be given to the professional development issues of junior faculty participating in interdisciplinary research teams. While participation may be "humbling" at times, the sheer volume of research progress that may be achieved through interdisciplinary collaboration, even in light of a short supply of grant dollars, is remarkable., (Copyright 2009 Elsevier Inc. All rights reserved.)

Prospects for discovering new antiparasitic drugs for veterinary medicine in the coming century will be determined by economic, social and scientific factors. Consolidation in the pharmaceutical industry in general, and the animal health industry in particular, changes the business conditions in which drug discovery for veterinary medicine occurs. Social pressures on traditional animal agriculture and companion animal ownership have shifted the interest of animal companies primarily to pet medicine. Antiparasitic drug discovery is more than ever targeted to the most lucrative market segments, but the excellence of available drugs, and the apparent lack of resistance in important parasites, reduces industrial motivation to invest in parasitology. Veterinary parasitologists in academia will still have the chance to interact with their industrial counterparts in the traditional ways of supporting drug discovery and development. Nonetheless, there are many new opportunities to expand the research horizons of veterinary parasitology to strengthen the case for retaining a significant presence in the animal health industry.

Beta-sialons are ceramic phases occurring in the SiO(2)-Si(3)N(4)-AlN-Al(2)O(3) system. A series of samples with differing compositions has been investigated by magic-angle spinning nuclear magnetic resonance (NMR) spectroscopy and powder X-ray diffraction (XRD). Although the constituent nitrogen nuclei occupy positions of low symmetry in the beta-sialon structure, 14N NMR spectra could be recorded for the samples examined. The origin of the 14N signal could be traced to the presence of an aluminium nitride (AlN) impurity phase with the help of 27Al NMR and XRD results. Similarly, the existence of Al(2)O(3) grains could be readily detected for a number of samples. Thus, the combination of 14N and 27Al NMR is shown to be an especially effective tool in identifying and characterizing impurity phases in sialon ceramics, complementing the results obtained from standard XRD analysis., (Copyright 2002 Elsevier Science (USA))

The organism about which most is known on a molecular level is a nematode, the free-living organism Caenorhabditis elegans. This organism has served as a reasonable model for the discovery of anthelmintic drugs and for research on the mechanism of action of anthelmintics. Useful information on mechanisms of anthelmintic resistance has also been obtained from studies on C. elegans. Unfortunately, there has not been a large-scale extension of genetic techniques developed in C. elegans to research on parasitic species of veterinary (or human) parasites. Much can be learned about the essentials of nematode biology by studying C. elegans, but discovering the basic biology of nematode parasitism can only be gained through comparative studies on multiple parasitic species.

To date, 9 FMRFamide-related peptides (FaRPs) have been structurally characterised from Caenorhabditis elegans. Radioimmunometrical screening of an ethanolic extract of C. elegans revealed the presence of two additional FaRPs that were purified by reverse-phase HPLC and subjected to Edman degradation analysis and gas-phase sequencing. Unequivocal primary structures for the two FaRPs were determined as Ala-Ala-Asp-Gly-Ala-Pro-Leu-Ile-Arg-Phe-NH(2) and Ser-Val-Pro-Gly-Val-Leu-Arg-Phe-NH(2). Using MALDI-TOF mass spectrometry, the molecular masses of the peptides were found to be 1032 Da (MH) and 875 Da (MH)(+), respectively. Two copies of AADGAPLIRFamide are predicted to be encoded on the precursor gene termed flp-13, while one copy of SVPGVLRFamide is located on flp-18. Synthetic replicates of the peptides were tested on Ascaris suum somatic muscle to assess bioactivity. ADDGAPLIRFamide had inhibitory effects on A. suum muscle strips, which occurred over a range of concentrations from a threshold for activity of 10 nM to 10 microM. SVPGVLRFamide was excitatory on A. suum somatic musculature from a threshold concentration for activity of 1 nM to 10 microM. The inhibitory and excitatory effects of AADGAPLIRFamide and SVPGVLRFamide, respectively, were the same for dorsal and ventral muscle strips as well as innervated and denervated preparations, suggesting that these physiological effects are not nerve cord dependent. Addition of ADDGAPLIRFamide (10 microM) to muscle strips preincubated in high-K(+) and -Ca(2+)-free medium resulted in a normal inhibitory response. Peptide addition to muscle strips preincubated in Cl(-)-free medium showed no inhibitory response, suggesting that the inhibitory response of the peptide may be chloride mediated. A normal excitatory response was noted following the addition of 10 microM SVPGVLRFamide to muscle strips preincubated in high-K(+), Ca(2+)- and Cl(-)-free media., (Copyright 2001 Academic Press.)

Two nitrogen ceramic phases, the oxynitride LiSiON and the nitride LiSi2N3, have been studied by 6Li and 7Li NMR. Magic angle spinning (MAS) NMR experiments have been carried out at two magnetic field strengths (7.05 and 14.1 T). The spectra give evidence of the relative effects of the quadrupolar and chemical shift interactions. The electric field gradient tensor of both phases has been determined accurately by iterative fitting of the 6Li and 7Li MAS NMR line shapes at the two magnetic field strengths. Due to the fact that for 7Li the quadrupolar interaction is much larger than the chemical shift interaction, it is shown that neither the small chemical shift anisotropy nor the relative orientation of the two interaction tensors can be determined accurately by 7Li MAS NMR. For 6Li, the two interactions are comparable and the value of these parameters obtained from the fits of the 6Li experimental MAS line shapes are therefore much more reliable.

Research in anthelmintic pharmacology faces a grim future. The parent field of veterinary parasitology has seemingly been devalued by governments, universities and the animal industry in general. Primarily due to the success of the macrocyclic lactone anthelmintics in cattle, problems caused by helminth infections are widely perceived to be unimportant. The market for anthelmintics in other host species that are plagued by resistance, such as sheep and horses, is thought to be too small to sustain a discovery program in the animal health pharmaceutical industry. These attitudes are both alarming and foolish. The recent history of resistance to antibiotics provides more than adequate warning that complacency about the continued efficacy of any class of drugs for the chemotherapy of an infectious disease is folly. Parasitology remains a dominant feature of veterinary medicine and of the animal health industry. Investment into research on the basic and clinical pharmacology of anthelmintics is essential to ensure chemotherapeutic control of these organisms into the 21st century. In this article, we propose a set of questions that should receive priority for research funding in order to bring this field into the modern era. While the specific questions are open for revision, we believe that organized support of a prioritized list of research objectives could stimulate a renaissance in research in veterinary helminthology. To accept the status quo is to surrender.

To date, 53 peptides with C-terminal RFamides have been identified by the genome sequencing project in the nematode, Caenorhabditis elegans. In this study the FMRFamide-related peptide (FaRP) KPSFVRFamide (879.90 Da [MH]+) was structurally characterized from extracts of the nematode, Caenorhabditis elegans. Two copies of KPSFVRFamide are encoded by a gene designated flp-9. RT-PCR identified a single cDNA product which was confirmed as flp-9 by sequence determination. Flp-9 cDNA was isolated from larval stages of C. elegans but was not detected in adult worms, indicating that its expression is may be developmentally regulated. KPSFVRFamide displays sequence homology to the nematode peptide, KPNFIRFamide (PF4). The physiological effects of KPSFVRFamide, PF4 and the chimeras, KPNFVRFamide and KPSFIRFamide, were measured on body wall muscle and the vagina vera of the parasitic nematode, Ascaris suum. KPNFVRFamide and KPNFIRFamide had Cl--dependent inhibitory activity on innervated and denervated muscle-preparations, whereas KPSFVRFamide and KPSFIRFamide did not elicit a detectable physiological effect. Although all 4 peptides had inhibitory effects on the vagina vera, KPSFVRFamide and KPSFIRFamide (threshold, >/=0.1 microM) were less potent than KPNFVRFamide and KPNFIRFamide (threshold, >/=10 nM)., (Copyright 1999 Academic Press.)

To date, seven FMRFamide-related peptides (FaRPs) have been structurally characterized from C. elegans, of which one is structurally identical to the parasitic nematode peptide AF2 (KHEYLRFamide). The other six FaRPs have so far been identified in free-living forms only. In the present study an additional FaRP was isolated and structurally characterized from an ethanolic extract of C. elegans. The extract was screened using a C-terminally directed FaRP antiserum, and the FMRFamide-immunoreactive peptide purified to homogeneity using HPLC. Approximately 80 pmol of the peptide was subjected to Edman degradation and the unequivocal primary structure of the K7-amide, KSAYMRFamide (PF3/AF8) was determined following a single gas-phase sequencing run. The molecular mass of the peptide was determined using a MALDI-TOF mass spectrometer and was found to be 919 (MH+), which is in agreement with the theoretical mass of C-terminally amidated PF3. A new flp-gene, designated flp-6, has recently been identified which encodes six copies of KSAYMRFamide (PF3/AF8)., (Copyright 1998 Academic Press.)

The effective concentrations of benzoic acid (BA), butylated hydroxyanisole (BRA), and carvacrol (CA) that reduced radial growth of Fusarium and Penicillium species by 50% (EC 50 ) are reported. The EC 50 of BA, BRA, CA was lower for species of Fusarium, ranging from 210 to 420 μg/ml of BA, 50 to 75 μg/ml of BRA, and 50 to 165 μg/ml of CA. The EC 50 for Penicillium species of BA, BRA, and CA ranged from 250 to 3,000 μg/ml, 100 to 275 μg/ml, and 200 to 350 μg/ml, respectively. While the test fungi showed varying responses to subinhibitory concentrations of the compounds, the results of this investigation revealed that Fusarium species were more sensitive to BA, BRA, and CA than Penicillium species.

To date, 9 FMRFamide-related peptides (FaRPs) have been identified in Caenorhabditis elegans. Eight of these peptides are encoded on the flp-1 gene. However, AF2 (KHEYLRFamide) which was not co-encoded was the most abundant FaRP identified in ethanolic extracts. Further radioimmunometrical screening of acidified ethanol extracts of C. elegans has revealed the presence of other novel FaRPs, which are not encoded on the flp-1 gene. One of these peptides has been isolated by sequential rpHPLC and subjected to Edman degradation analysis and gas-phase sequencing and the unequivocal primary structure of the decapeptide Ala-Pro-Glu-Ala-Ser-Pro-Phe-Ile-Arg-Phe-NH2 was determined following a single gas-phase sequencing run. The molecular mass of the peptide was found to be 1133.7 Da, determined using a time-of-flight mass spectrometer. Synthetic replicates of this peptide were found to induce a profound relaxation of both dorsal and ventral somatic muscle-strip preparations of Ascaris suum with a threshold for activity of 10 nM. The inhibitory response was not dependent on the presence of nerve cords, indicating a post-synaptic site-of-action. The relaxation was Ca(+2)- and Cl(-)-independent but was abolished in high-K+ medium and could be distinguished from those of other inhibitory nematode FaRPs, including PF1 (SDPNFLRFamide) and PF4 (KPNFIRFamide).

The antifungal effect of butylated hydroxyanisole (BHA) and carvacrol alone and in combination on six toxigenic Fusarium species was examined in potato dextrose broth (PDB) cultures. The highest percentage of mycelial inhibition was recorded for F. poae in the presence of 200 μg/ml of BHA or carvacrol and BHA and carvacrol combined (50 μg/ml each and 100 μg/ml each), while F. moniliforme was the least sensitive of the test fungi at the same concentrations. A synergistic activity of BHA and carvacrol combined (50 μg/ml each) was observed against several of the test fungi. The pH of the filtrates from BHA or carvacrol and BHA plus carvacrol-amended cultures ranged between 5.0 and 5.4. The highest amount of total dissolved solids (TDS) was recorded for F. graminearum in the presence of 200 μg/ml of BHA. Leakage of amino acids, sugars, and protein was detected at each concentration of BHA or carvacrol alone and in combination. For the majority of the test fungi, the amount of leakage in the culture filtrates increased as the concentration of the test compounds increased. The leakage of sugars, amino acids, and protein may be an indication of mycelial membrane disruption by the BHA and/or carvacrol as an initial step in inhibition of the test fungi in vitro.

Numerous FMRFamide-related peptides (FaRPs) have been isolated and sequenced from extracts of free-living and parasitic nematodes. The most abundant FaRP identified in ethanolic/methanolic extracts of the parasitic forms, Ascaris suum and Haemonchus contortus and from the free-living nematode, Panagrellus redivivus, was KHEYLRFamide (AF2). Analysis of the nucleotide sequences of cloned FaRP-precursor genes from C. elegans and, more recently, Caenorhabditis vulgaris identified a series of related FaRPs which did not include AF2. An acid-ethanol extract of Caenorhabditis elegans was screened radioimmunometrically for the presence of FaRPs using a C-terminally directed FaRP antiserum. Approximately 300 pmols of the most abundant immunoreactive peptide was purified to homogeneity and 30 pmols was subjected to Edman degradation analysis and gas-phase sequencing. The unequivocal primary structure of the heptapeptide, Lys-His-Glu-Tyr-Leu-Arg-Phe-NH2 (AF2) was determined following a single gas-phase sequencing run. The molecular mass of the peptide was determined using a time-of-flight mass spectrometer and was found to be 920 (MH+)+, which was consistent with the theoretical mass of C-terminally amidated AF2. These results indicate that C. elegans possesses more than one FaRP gene.

This multidisciplinary study demonstrates the utility of the biophysical model approach to assess biological activity of anthelmintics in light of drug-delivery principles. The relationships between drug absorption and efficacy for a set of structurally disparate anthelmintics were determined in cultures of Haemonchus contortus, a nematode that parasitizes the ruminant gastrointestinal tract. Uptake, parameterized by the permeability coefficient, Pe, was shown to occur by absorption across the cuticle. Rates of drug appearance in nematode carcasses paralleled rates of drug disappearance from the medium, and absorption reached an apparent equilibrium within a few hours. The parasite/medium partition coefficient, K, was derived from the ratio of drug concentration in the parasite vs the medium at equilibrium. Pe and K values for each anthelmintic were correlated with lipophilicity (as measured by the partition coefficient (PC) in n-octanol/water) and both parameters plateaued at log PC approximately 2.5, with maximum Pe approximately 8 x 10(-4) cm/min and log K < or = 2.0. Absorption kinetics were related to in vitro potency by monitoring motility of H. contortus. The time required to reduce motility by 50% (t* 50) and Pe were used to calculate Cn*, the drug concentration in the parasite at t* 50, as an indicator of intrinsic potency. The quantitative interplay of apparent biological activity expressed as t* 50, dose, and intrinsic potency highlights the important contribution of drug-uptake kinetics.

In nematodes, FMRFamide-related peptides (FaRPs) have been structurally characterised from the parasite, Ascaris suum, and from two free-living species, Panagrellus redivivus and Caenorhabditis elegans. While both FaRPs isolated from P. redivivus (PF1 and PF2) have been identified in C. elegans, the two heptapeptides isolated from A. suum (AF1 and AF2) have until recently been considered unique to this parasitic species. We have recently isolated AF2 from P. redivivus and, during this study, an additional novel heptapeptide amide, Lys-Ser-Ala-Tyr-Met-Arg-Phe amide (KSAYMRFamide), was structurally characterised. A synthetic replicate of this peptide induced a rapid concentration-dependent muscle tension increase in an isolated. A. suum somatic muscle preparation, with a threshold of approximately 0.1 microM. These data suggest that the complement of FaRPs in parasitic and free-living nematodes may not be as radically different as preliminary studies would suggest, and that the absence of AF1, AF2 and KSAYMRFamide on the C.elegans FMRFamide-related peptide gene (flp-1) may imply the presence of at least two different FaRP genes in nematodes.

The effect of pH alone and in combination with two phenolic antioxidants on mycelial growth and conidial germination of mycotoxigenic fungi was examined. The mycotoxigenic fungi studied included seven members each of Fusarium and Penicillium genera. The present study was carried out at pH 4, 6, 8, and 10 alone and in the presence of 100 and 200 μg/ml of butylated hydroxyanisole (BHA) and propyl paraben, respectively. All of the test fungi examined were able to grow at pH 4, 6, 8, and 10 on potato dextrose agar. The majority of the Fusarium species, with the exception of Fusarium graminearum , were tolerant of acid and alkaline pHs. Two of the Penicillium species were rapid growers and were tolerant of acid and alkaline pHs. The tolerance of the slower growing Penicillium species varied in their response to acid and alkaline pHs. Conidial germination was partially inhibited in all of the Fusarium and Penicillium species in the presence of 100 μg/ml of BHA at all pH levels. However, in the presence of 200 μg/ml of BHA, there was complete inhibition in conidial germination for four of the Fusarium species at pH 4, 6, 8, and 10. Propyl paraben, at the four pH levels, completely inhibited conidial germination in Fusarium species and all but one species of Penicillium at 200 μg/ml. Reduction in colony diameter for the majority of Fusarium and Penicillium species, at all pH levels was observed at 100 and 200 μg/ml of BHA. However, at 200 μ/ml of Propyl paraben, mycelial growth was completely inhibited for the majority of the Fusarium and Penicillium species at the four pH levels.

The fungitoxic effect of butylated hydroxyanisole (BHA) on germination of conidia from seven toxigenic strains of Aspergillus was examined on potato dextrose agar (PDA). At concentrations of 200 μg/ml and above, BHA prevented germination of conidia in all of the test fungi after 24 and 48 h of incubation. On the other hand, at 100 μg/ml BHA reduced the rate of germination. When an impregnated disk assay was employed, distinct inhibition zones were observed at 200, 400, 800, and 1000 μg/disk of BHA after 24 h of incubation. After 48 h of incubation, distinct inhibition zones were still observed at 800 and 1000 μg/disk.

The optimum pH for growth for eight toxigenic strains of Aspergillus was recorded at pH 6 in unamended potato dextrose broth medium. Significantly less mycelial growth was observed at pH 4 and 8, respectively, for all of the fungal strains. In the presence of 1.0 mM of thymol, mycelial growth was completely inhibited at pH 4, 6, and 8 for all of the fungal strains. Carvacrol, at the same concentration, completely inhibited the growth of the fungi only at pH 4 and 8. At lower concentrations (0.1 and 0.5 mM), in the presence of thymol or carvacrol, partial mycelial inhibition at all pH levels was observed with significantly less mycelial growth at pH 4 and 8.