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.     

Development of resistance to Trichostrongylus colubriformis in goats

A high level of host resistance to Trichostrongylus colubriformis was established in a group of Saanen goats infected weekly with 10,000 infective larvae for 10 weeks, treated with an anthelmintic and then challenged twice with 50,000 larvae, with a further anthelmintic treatment 28 days after the first challenge. A second group of goats exposed only to the two challenge infections developed worm burdens intermediate between, and significantly different from, the first group and a third control group given only the second challenge infection. Nematode fecundity (eggs per female) and male:female ratios were also significantly decreased in the first two groups. Faecal egg counts and worm counts were highly correlated and this relationship was dominated by the influence of the number of eggs/female nematode. The goats were approximately 9 months old at the start of the experiment.     

The use of PCR to detect Neospora caninum DNA in the blood of naturally infected cows

Twelve 2-year old heifers in their fifth month of gestation when pregnancy tested were used in this study. Six heifers aborted at approximately 4 months of gestation and had blood samples drawn less than 6 weeks after the abortions were identified. Blood samples were also drawn from three sero-positive pregnant and three sero-negative pregnant heifers. DNA was isolated from the samples and a 350 bp fragment of the Nc-5 gene was PCR amplified using primer pair Np21+ and Np6+. Also, the Internal Transcribed Spacer 1 (ITS1) was PCR amplified using Tim 3 and Tim 11 primer pair. The Nc-5 gene fragment was cloned, sequenced and the sequence BLAST-tested. Similarly, the ITS1 product was sequenced and BLAST-tested. The BLAST test results revealed that Neospora caninum DNA was present in these blood samples indicating that polymerase chain reaction can be used in the detection of N. caninum DNA in the blood of sero-positive cows.     

Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates. METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent antibody test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice. RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3-7%. CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

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.     

Farm management practices associated with macrocyclic lactone resistance on sheep farms in New Zealand

AIM: To identify farm practices associated with the presence of resistance to a macrocyclic lactone (ML) anthelmintic on sheep farms in New Zealand. METHODS: A cross-sectional study was conducted to test for associations between the presence of resistance to an ML anthelmintic (ivermectin) and management practices on sheep farms in New Zealand. Selection of farms was both random (n=80) and purposive (n=32; being farms with a history of suspected ML resistance). Resistance was inferred from faecal nematode egg count (FEC) reduction (FECR) tests (FECRTs) when there was <95% reduction in FEC 7-10 days after treatment with a half dose of ivermectin (0.1 mg/kg). A logistic regression model was built to identify farm-level factors that were associated with the presence or absence of ML resistance. RESULTS: Of the 112 flock managers that were approached for interview, 103 (92%) returned useable questionnaires. The odds of ML resistance were increased: on farms that had used long-acting ML products in ewes as a pre-lambing treatment for > or =3 of the previous 5 years (odds ratio (OR) = 7.2; 95% confidence interval (CI) = 1.7-30.3); on farms where <70% of the total stock units mid-winter were from sheep (OR=6.5; 95% CI=1.6-25.6); on farms which over the year purchased >10% of the number of sheep present mid-winter (OR=7.1; 95% CI=1.5-34.7); and on farms where the average wool diameter of the main flock was <37 (OR=4.1; 95% CI=1.1-14.7) microns. The model provided a good fit to the data (pseudo R2=0.64; Hosmer-Lemeshow statistic = 0.38). CONCLUSIONS: Explanatory factors identified as associated with the presence of ML (ivermectin) resistance on farms included the use of long-acting anthelmintic formulations in ewes pre-lambing, sources of refugia of unselected parasites on the farm, breed of sheep and their requirements for anthelmintic treatments, and the importing of resistant parasites with purchased stock. The study provides support for controls that aim to provide refugia of susceptible worms and that minimise the risk of introduction of resistance through effective quarantine drenching.     

Anthelmintic resistance and management of nematode parasites on beef cattle-rearing farms in the North Island of New Zealand

AIM: To provide information on current farmers' opinions and farming practices thought to be related to anthelmintic resistance, and to test for associations between the presence of anthelmintic resistance and management practices on beef-cattle rearing farms in the North Island of New Zealand. METHODS: A study using an interview-based questionnaire about management of internal parasites was conducted on 62 beef cattle-rearing farms in the North Island of New Zealand, using case-control analyses to test for associations between management practices and the presence or absence of resistance to ivermectin or albendazole. Resistance was inferred from faecal nematode egg count (FEC) reduction (FECR) tests (FECRTs) when there was <90% reduction in FEC 7-10 days after treatment of calves <12 months of age. RESULTS: Of the 59 farmers who completed the questionnaire, most (n=40) ranked parasites highly, and at about the same level as quality and quantity of feed, as important production-limiting factors for their enterprises. In contrast, anthelmintic resistance was not perceived to be a problem on 13 farms, and its importance was rated low on 24, moderate on 15, and high on only six farms. Despite all farms having planned parasite control programmes, there was heavy reliance on clinical signs of parasitism to determine frequency of treatments. About one in three farmers with beef breeder cows routinely treated their calves at marking, one in five treated mixed-age cows, and almost half treated rising 2-year-old cows before calving. One in four farmers used anthelmintics on calves on 8-12 occasions in their first year of life. Co-grazing with other species was rare, but follow-on grazing within 3 months after older cattle or sheep was common. On most farms, grazing cattle was restricted to part of the farm, a finding with implications for parasite control and persistence of larvae in refugia. Macrocyclic lactone (ML) anthelmintics or their combinations with other action families were currently, and for the past 5 years, used more frequently than benzimidazoles and levamisole, and benzimidazole-levamisole combinations. The prevalence of resistance to ivermectin was high (82%) and no plausible model of associations could be constructed from the data. The prevalence of resistance to albendazole was 60%, and the risk of resistance increased as the number of rising 1-year-old cattle present mid-winter increased, and decreased as the number of breeding cows >2 years old present mid-winter increased. CONCLUSION: It is clear that in practice anthelmintic resistance is a secondary consideration to obtaining productivity advantages from the use of anthelmintics in beef cattle. Farmers' opinions were divided on many issues and the overall impression was of confused and diverse thinking regarding the principles of the use of anthelmintics. The overall outlook regarding anthelmintic resistance in cattle is bleak unless the need for integrated and long-term research activities is acted upon soon.     

The effect of repeated, four-weekly eprinomectin treatment on milk production in pasture-based, seasonally-calving dairy cattle

A randomised clinical trial from the North Island of New Zealand was conducted to assess the effect of repeated anthelmintic treatment on milk production, and to assess factors that affect treatment response. Nine hundred and twenty three multiparous, lactating dairy cattle from three pasture-based, spring-calving dairy herds were enrolled in this trial. Within each herd, cattle were stratified on age and calving date, and were randomly allocated to treatment (n=319) or control (n=604) groups. The treatment group received ≥0.05mg/kg of topical eprinomectin every 28 days for eight treatments during lactation. Pooled-milk from treated cows and bulk-milk samples were obtained at each treatment and analysed with an Ostertagia antibody ELISA, expressed as optical density ratios (ODR). Bi-monthly milk data were collected and expressed as energy-corrected milk (kg/day; ECM). A linear mixed model was used to analyse ECM, with cow as the random effect. The effect of anthelmintic treatment on days from calving, and start-of-mating, to conception were analysed with Cox-proportional hazard models. ODR values ranged from 0.6 to 1.3; there were no differences in ODR between herds (p=0.12), or between pooled-milk from treated cows and bulk-milk (p=0.26). Repeated treatments had no effect on daily ECM yields (p=0.74). However, there was a significant treatment×herd interaction (p=0.03); treatment increased ECM in one herd by 0.781kg/cow/day (p=0.015), but resulted in a non-significant decrease in the other two herds. A curvilinear interaction existed between days-in-milk and treatment response (p=0.039); the greatest treatment effect occurred during mid-lactation. Previous year milk production (p=0.46) and age (p=0.11) did not influence the effect of treatment on ECM. Treatment had no effect on any reproductive parameter. In conclusion, under New Zealand pastoral conditions, anthelmintic treatment increased milk production in one herd, but had no effect in two other herds. Further work is needed to identify why this variation in gastro-intestinal parasitism occurs.     

Computed tomographic assessment of vascular invasion and resectability of mediastinal masses in dogs and a cat

AIMS: To assess the sensitivity of non-angiographic contrast-enhanced computed tomography (CT) to determine the presence of vascular invasion of cranial mediastinal masses in dogs and a cat, and to evaluate the association between vascular invasion and peri-operative mortality.

METHODS: A retrospective study was conducted on 25 dogs and one cat. CT scans were completed with slices ranging from 2 to 10 mm. CT images were evaluated by a board-certifi ed radiologist blinded to previous diagnoses and surgical fi ndings. Each CT study was evaluated for vascular invasion, defi ned as disruption of the vessel wall and extension of the mass into the vessel lumen. Data retrieved from the surgery reports included surgical approach, whether vascular invasion was present, the surgeon's decision on operability, and post-operative complications.

RESULTS: Computed tomographic evaluation revealed 25/26 masses had no evidence of vascular invasion. During surgical exploration, 10/26 masses were found to invade major regional vasculature; the cranial vena cava (CVC) was the vessel most commonly invaded (7/10 animals), and 4/7 (57%) patients with invasion of the CVC were euthanised or died in the perioperative period, from surgical or disease-related problems, which was signifi cantly higher than patients without vascular invasion (p=0.045).

CONCLUSIONS: Non-angiographic contrast-enhanced CT was signifi cantly less sensitive for detecting vascular invasion of cranial mediastinal masses when compared with surgical evaluation. If the CVC was invaded by a tumour there was a signifi cant risk of death peri-operatively when compared with non-invasive cases.

CLINICAL RELEVANCE: Due to the signifi cantly higher mortality risk associated with invasion of the CVC, a more sensitive method than CT should be investigated to determine vascular invasion of mediastinal masses pre-operatively.