In vitro effects of DDT analogs on trout brain Mg2+-ATPases: Inhibition kinetics

A continuous in vitro steady-state assay procedure was used to investigate the dependence of trout brain mitochondrial Mg²⁺-stimulated adenosinetriphosphatase specific activity on temperature and substrate concentration. The inhibition of enzyme activity by DDT was independent of substrate concentration. DDT and several analogs caused increases in the experimental activation energy and frequency factor of the enzyme-catalyzed reaction, which gave rise to a negative temperature coefficient of inhibition. It is suggested that DDT and other highly lipophilic compounds have the potential to allosterically affect membrane-bound enzymes by simply becoming a major lipoid component of the membranes.     

Evaluation of formalin-fixed paraffin-embedded tissues from feline vaccine site-associated sarcomas for feline foamy virus DNA

OBJECTIVE: To evaluate a group of vaccine site-associated sarcomas (VSS) for the presence of feline foamy virus (FeFV) DNA, using polymerase chain reaction (PCR) methods. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded (FFPE) tissue blocks from VSS of cats. PROCEDURE: DNA was extracted from FFPE sections of each tumor, and regions of the gag and pol genes of FeFV were amplified by use of PCR methods, using 1 primer set for each region. Sensitivity of the method was compared between fresh and FFPE cells, using mouse kidney tissue that was injected with FeFV-infected cultured cells and using agarose-cell pellets. Results-Feline foamy virus DNA was not detected in VSS tissues. Sensitivity of the method was 10 times greater in fresh versus FFPE mouse tissues. Sensitivity of the method in fresh FeFV-infected cultured cells versus FFPE agarose-cell pellets was equal when fixation was 24 or 48 hours and 10 times greater when fixation was 72 hours or 1 week. CONCLUSIONS AND CLINICAL RELEVANCE: A PCR-based method can be successfully applied to FFPE tissues for FeFV DNA detection. Results suggest there is no direct FeFV involvement in the pathogenesis of VSS in cats.     

Reproductive performance of beef cow herds in New Zealand

AIM: To describe the reproductive performance of beef cow herds in New Zealand and to develop reference ranges for assessing the reproductive performance of individual herds from in-calf rates, that take into account variation in the length of mating periods. METHODS: Veterinary practices throughout New Zealand involved in beef cattle work were asked to collect reproductive data from seasonally calving beef cow herds mated in the spring of 2001 through to the end of summer of 2002. An estimate of conception rate (termed calculated conception rate: CCR) was determined for each herd, assuming that the conception rate was constant for each 21-day interval of the mating period. The algebraic relationship between CCR and in-calf rate at pregnancy testing was defined for mating periods of different durations and, therefore, given the in-calf rate and the duration of the mating period, a CCR could be determined for each herd. Expected pregnancy rates were recalculated from CCR data for a range of mating period durations to produce a look-up table for assessing herd reproductive performance. Reproductive data describing regional differences in in-calf rates and CCRs, bull:cow ratios, breed characteristics, start dates of mating and durations of mating periods were summarised. The effect of study variables in explaining CCRs was examined using a general linear model (GLM). RESULTS: Data were collected from 1,005 beef cow herds distributed throughout New Zealand. The median in-calf rate for all herds was 91%, the lower quartile was < or =88% and the upper quartile > or =94%. The mean CCR for herds with complete reproductive data (862) was 55% (SD 11), the lower quartile was < or =48% and upper quartile > or =61%. Median in-calf rates for 2-year-old heifers (mated at approximately 15 months of age), 3-year-old heifers (mated at approximately 27 months of age), and mixed-age cows were 90%, 91% and 92%, respectively. The study variables that accounted for significant variation in breeding group CCR in a multivariate GLM were 'region' (p<0.01) and 'date mating commenced' (<0.01). The adjusted R2 for the model was 0.055. CONCLUSIONS: The reproductive reference range produced provides veterinarians and herd managers with a quantitative method for assessing reproductive performance of beef cow herds compared with industry averages, from in-calf rates at the time of pregnancy testing and durations of mating periods.     

Evaluation of the effectiveness of Duddingtonia flagrans and Monacrosporium thaumasium in the biological control of gastrointestinal nematodes in female bovines bred in the semiarid region

Brazil has a herd of 212 million cattle and 171 million hectares of pastures that produce approximately 96 % of Brazilian beef. The Brazilian production system enables animal infection by endoparasites, which are considered one of the main obstacles for the development of this industry and are responsible for considerable economic losses. The control of parasitic diseases is performed via the administration of antiparasitic drugs, but they leave residues of the products in the treated animal, affect non-target organisms and select resistant strains of the parasites. The species D. flagrans and M. thaumasium are promising and sustainable alternatives for controlling gastrointestinal helminths of ruminants and other herbivores. In this study, we evaluated the efficacy of isolates of these species, formulated in a sodium alginate matrix and administered twice a week, to reduce the number of environmental infective larvae of gastrointestinal nematodes that affect prepubescent zebu females. The treated animals presented fewer eggs and a lower number of infective larvae per gram of faeces (p?<?0.05). The pastures occupied by treated animals showed a statistically significant reduction (p?<?0.05) of the number of L₃ and, furthermore, the genera Cooperia sp., Haemonchus sp., and Oesophagostomum sp. were the most prevalent. The average weight of the animals did not differ statistically (p?>?0.05) among the treated and control groups. The use of sodium alginate pellets as vehicle for delivery of the fungus mycelia D. flagrans (isolate AC001) and M. thaumasium (isolate NF34A) proved effective in controlling trichostrongylids in prepubescent cows bred in the semi-arid region, with an effective reduction in the number of infective larvae in the pastures.     

Pseudomonas syringae pv. coryli, the Causal Agent of Bacterial Twig Dieback of Corylus avellana

ABSTRACT Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by a polyphasic approach. All strains were assessed by fatty acids analysis and repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX and ERIC primer sets. Representative strains also were assessed by sequencing the 16S rDNA and hrpL genes, determining the presence of the syrB gene, testing their biochemical and nutritional characteristics, and determining their pathogenicity to hazelnut and other plants species or plant organs. Moreover, they were compared with reference strains of other phytopathogenic pseudomonads. The strains from hazelnut belong to Pseudomonas syringae (sensu latu), LOPAT group Ia. Both fatty acids and repetitive-sequence-based PCR clearly discriminate such strains from other Pseudomonas spp., including P. avellanae and other P. syringae pathovars as well as P. syringae pv. syringae strains from hazelnut. Also, the sequencing of 16S rDNA and hrpL genes differentiated them from P. avellanae and from P. syringae pv. syringae. They did not possess the syrB gene. Some nutritional tests also differentiated them from related P. syringae pathovars. Upon artificial inoculation, these strains incited severe twig diebacks only on hazelnut. Our results justify the creation of a new pathovar because the strains from hazelnut constitute a homogeneous group and a discrete phenon. The name of P. syringae pv. coryli is proposed and criteria for routine identification are presented.     

Prevalence of anthelmintic resistance on sheep farms in New Zealand

AIM: To establish the prevalence of anthelmintic resistance in parasitic nematodes on sheep farms in New Zealand. METHODS: A cross-sectional prevalence study was conducted, using a standardised faecal nematode egg count (FEC) reduction (FECR) test (FECRT) for ivermectin, at a full (0.2 mg/kg) and half (0.1 mg/kg) dose rate, and albendazole, levamisole and albendazole-levamisole in combination, on 60 lambs (n=10 per group) on farms selected from throughout New Zealand. Farms that conformed with selection criteria were chosen at random (n=80) or with a history of suspected resistance to macrocyclic lactone (ML) anthelmintics (n=32). Resistance to an anthelmintic was inferred when there was <95% reduction in FEC 7-10 days after treatment. Larval cultures were performed for all control groups and for treated groups for which resistance was evident. RESULTS: Of the farms randomly selected, 36% showed > or =95% FECR for all anthelmintics tested; resistance to ivermectin at 0.1 and 0.2 mg/kg liveweight was evident on 36% and 25% of these farms, respectively. Resistance to both ivermectin (0.2 mg/kg) and levamisole was evident on 8/80 (10%) farms, to ivermectin and albendazole on 10/80 (13%) farms, and to ivermectin, levamisole and albendazole on 6/80 (8%) farms. The prevalence of resistance to a half dose of ivermectin tended to be more prevalent on farms with a history of suspected ML resistance (p=0.06). Resistance to albendazole was seen across all the main parasite genera, and to levamisole in Nematodirus, Ostertagia (= Teladorsagia) and Trichostrongylus species. Resistance to ivermectin was dominated by Ostertagia spp, although Cooperia, Nematodirus and Trichostrongylus species were also implicated. CONCLUSION: Anthelmintic resistance in parasitic nematodes of sheep is common in New Zealand. Not only was resistance to albendazole and levamisole common, but resistance to the ML, ivermectin, was at a higher prevalence than expected. Sheep farmers and advisors in New Zealand need to re-evaluate the way they manage parasites, and more research is urgently needed if the steady decline in anthelmintic susceptibility is to be halted.     

Drug sales and the veterinarian

Sir,—Most private practitioners and probably all veterinary clubs in New Zealand derive a considerable portion of their incomes from the sales of drugs, and it is certainly true that most veterinary dubs are dependent on drug sales profits to allow them to continue at their present economic levels.     

Detection and measurement of benzimidazole resistance alleles in Haemonchus contortus using real-time PCR with locked nucleic acid Taqman probes

Benzimidazole resistance is a common problem in parasitic nematodes of ruminants and early detection is vital if its spread is to be monitored and controlled. Real time PCR offers a fast and reliable method for rapid detection and measurement of resistance allele frequencies. In Haemonchus contortus a single nucleotide polymorphism at codon 200 of the beta-tubulin gene (TTC to TAC), causing a phenylalanine to tyrosine amino acid substitution, has been shown to be involved in many cases of resistance. Locked nucleic acid (LNA) Taqman probes have been used in this work to detect and measure the frequency of resistance alleles in individual and multiple H. contortus. Detection of resistant genotypes using LNA Taqman probes in individual H. contortus is simpler and more reliable than with previously described assays. Measurement of the frequency of resistant alleles in populations of H. contortus was achieved by using the cycle threshold (C(t)) values and a standard curve derived from populations with known allele frequencies. Results using the LNA probes on individual and multiple worms gave similar results to the allele specific PCR. The sensitivity of the LNA assay on multiple nematodes allowed reliable detection of > or = 10% resistance allele frequency. Using the final fluorescence method, it was possible to differentiate populations with approximately 0, 5 and 10% resistance allele frequencies.     

酸度对胡敏酸与镉和锌离子络合反应的影响

用离子交换平衡法研究了不同pH下胡敏酸与锌、镉单独及复合存在下形成络合物的稳定性及配位数.结果表明:在Zn2+,Cd2+单独存在时,络合反应的稳定常数及配位数随着pH值的增大而增大;相同pH条件下,Zn2+和胡敏酸络合物的稳定常数及配位数大于Cd2+.在Zn2+,Cd2+复合存在时,pH的变化与胡敏酸络合金属离子的稳定常数及配位数没有明显依存关系;与单独存在时相比,在pH 3.5和pH7.0时,Zn2+,Cd2+和胡敏酸络合物的稳定常数和配位数都减小;而在pH 5.0时,Cd2+和胡敏酸络合物的稳定常数和配位数都增大,Zn2+和胡敏酸络合物的稳定常数和配位数都减小.