Evaluation of the infectivity of Trichinella papuae and Trichinella zimbabwensis for equatorial freshwater fishes

The discovery of Trichinella species infecting poikilotherm vertebrates has opened new possibilities in the epidemiology of this parasite group. The aim of the present work was to investigate the infectivity of the two non-encapsulated species of Trichinella infecting both mammals and reptiles, Trichinella papuae and Trichinella zimbabwensis, for equatorial freshwater carnivore fishes. To this end, two species of piranhas, four Serrasalmus nattereri and four Serrasalmus rhombeus, were each inoculated per os with the two species of Trichinella larvae. Six days post infection (p.i.), one fish of each species inoculated with one of the two species of Trichinella was sacrificed. The intestines and celomatic cavities were searched for worms using dissection microscopy, and the presence of muscle larvae was evaluated by artificial digestion. The other 4 inoculated fish were sacrificed 60 days p.i. and similarly searched for the presence of worms. No larva or adult worms were detected in any organ or tissue at 6 or 60 days p.i. The lack of infectivity of T. papuae and T. zimbabwensis for fish suggests that the entozoic habitat of this animal does not represent a suitable environment for these two Trichinella species. More importantly, these data indicate that freshwater fishes, one of the food resources for crocodiles, caimans and alligators, are unlikely to play a role in the epidemiology of the known species of the genus Trichinella.     

半滑舌鳎弧菌病病原的分离鉴定

从患病半滑舌鳎(Cynoglossus semilaevis)内脏中分离到2株优势菌株,人工感染试验证实菌株150803对半滑舌鳎具有致病性;采用形态学观察、生理生化特性分析、16S r RNA等方法对所分离菌株进行鉴定,结果显示,菌株150803为革兰阴性杆菌,生化特性与溶藻弧菌(Vibrio alginolyticus)较为接近,以hsp60基因为遗传标记构建的系统发育树将菌株150803与溶藻弧菌聚为一支,置信度为99%,采用特异性引物扩增溶藻弧菌dnaJ基因得到预期144 bp的基因片段。综合以上结果判定引起此次半滑舌鳎发病的病原为溶藻弧菌。对22种抗菌药物的敏感性试验表明,菌株150803对恩诺沙星、环丙沙星、庆大霉素、菌必治敏感,对氟苯尼考、呋喃唑酮、苯唑青霉素等具有抗性。     

Evaluation of a mixture of trimethoprim and sulphaquinoxaline for the treatment of bacterial and coccidial diseases of poultry

Representative experiments from work undertaken to develop a synergistic mixture of trimethoprim and sulphaquinoxaline for the preventive treatment of certain poultry diseases are described. Sulphaquinoxaline in the diet for four days was shown to achieve at least an 85 per cent higher blood level than nine other sulphonamides in chicks, and the efficacies of various trimethoprim/sulphaquinoxaline regimes in the diet or in the drinking water were demonstrated against pasteurellosis, colisepticaemia and five kinds of coccidiosis. Regimes for bacterial diseases were begun one day before infection but those for coccidial diseases were begun on the same day as infection or later. Overall, a total dose of 30 mg/kg bodyweight/day (trimethoprim/sulphaquinoxaline = 1:3) controlled these seven diseases. The same treatment was also shown to control sulphaquinoxaline-resistant strains of Escherichia coli and Eimeria acervulina. Although both drinking water and food were used for drug administration, twice the inclusion rate was required in food to that in water for equivalent efficacy. The significance of different modes of expression of dosages for bacterial and coccidial diseases is explained.     

The development and analysis of species specific and cross reactive monoclonal antibodies to leukocyte differentiation antigens and antigens of the major histocompatibility complex for use in the study of the immune system in cattle and other species

We examined the potential of developing a set of species specific and cross reactive monoclonal antibodies (MoAbs) for use in the study of the phylogenetic and functional relation of class I and class II antigens of the major histocompatibility complex (MHC) and leukocyte differentiation antigens in cattle and other species. Comparing immunization strategies demonstrated the number of hybrids producing cross reactive antibodies can be increased by hyperimmunization of mice with lymphoid cells from multiple species. Comparing various methods of assay (antibody-complement mediated cytotoxicity [CT], enzyme linked immunosorbent assay [ELISA] and flow microfluorimetry [FMF]), revealed FMF is the most useful technique for the primary assay of hybridomas producing MoAbs of potential interest. By using dual parameter and dual fluorescence analysis, we could determine whether a given MoAb reacted with mononuclear cells (lymphocytes and monocytes) and/or granulocytes, and also whether any two MoAbs of different isotype and specificity recognized antigens present on identical or separate populations of leukocytes. Comparing the patterns of MoAb reactivity with leukocytes obtained from cows, goats, sheep, pigs, horses and humans, as well as comparing the patterns of reactivity with a panel of lymphoid cell lines derived from cattle (with enzootic bovine leukemia) and humans (with various forms of leukemia), revealed sets of MoAbs reactive with unique antigenic determinants present on BoLA class I (15 MoAbs) and class II (9 MoAbs) antigens, and also MoAbs reactive with determinants present on leukocyte differentiation antigens (36 MoAbs). Dual fluorescence analysis demonstrated the antigens detected by some MoAbs are predominantly expressed on one lineage of leukocytes while others are expressed on two or more lineages of leukocytes. Dual and single fluorescence analysis also demonstrated the PNA receptor(s) is: expressed on T cells, granulocytes and class II antigen monocytes and absent or expressed in low amount on sIgM+ B cells and a newly defined Non T/Non B population of cells. The strategies described for identifying and analyzing the specificity of MoAbs demonstrate the feasibility of developing a set of cross reactive MoAbs for identifying homologous molecules in multiple species and delineating their functional and phylogenetic relation.     

Susceptibility of different bacterial species isolated from food animals to copper sulphate, zinc chloride and antimicrobial substances used for disinfection

A total of 569 different bacterial isolates (156 Salmonella, 202 E. coli, 43 S. aureus, 38 S. hyicus, 52 E. faecalis, 78 E. faecium) were tested for susceptibility to copper sulphate, benzalkonium chloride, hydrogen peroxide and chlorhexidine using MIC determinations. A total of 442 isolates were also tested for susceptibility to formaldehyde and 177 isolates for susceptibility to zinc chloride. Enterococcal isolates formed a bimodal distribution of MICs to copper sulphate, whereas the other bacterial species formed one large population. Otherwise the isolates formed one large population of susceptibilities to the different antimicrobial agents. Large variations were observed in the susceptibility of the different bacterial species to the different compounds. Staphylococci were in general very susceptible to all antimicrobial compounds tested. The Salmonella isolates were in general less susceptible to copper sulphate, benzalkonium chloride and chlorhexidine followed by E. coli and the Gram-positive species. The opposite was the case for zinc chloride. All isolates were very susceptible to H(2)O(2) with MICs ranging from 0.002 to 0.016%, and to formaldehyde with MICs at 0.003 and 0.006%. This study showed that Danish bacterial isolates from livestock so far have not or have only to a limited degree developed resistance to antimicrobial compounds commonly used for disinfection. Acquired copper resistance was only found in enterococci. There were large differences in the intrinsic susceptibility of the different bacterial species to these compounds, and Salmonella especially seems intrinsically less susceptible than the other bacterial species, which might have human health implications.     

蚕桑生产下滑原因及对策

1　我区蚕业现状我区的蚕业生产在 90年代前都保持持续发展的趋势 ,1 990年养蚕 2 .1万张 ,首次突破产茧 50 0t大关 ,继后四年中 ,产茧均以 2 50t速度递增 ,1 994年养蚕 4 .6万张 ,产茧 1 50 0t,茧款收入达 1 80 0万元 ,达到了我区蚕业史上的最高峰 ,涌现出产茧 1 50t的乡 1个 ,1 0 0t的乡 2个 ,50t乡 1个 ;养蚕收入 1 0～ 1 5万元的专业村 3个 ;1～ 2万元的大户 2个 ;曾出现过市劳模养蚕状元袁天奇 ,年养蚕 56张 ,收入 2 .8万余元而远近闻名。以后随着市场的变化 ,茧价下降等原因 ,全区挖桑毁桑特别严重 ,养蚕逐年减少 ,至 2 0 0 0年全区养…     

Using indirect ELISA to assess different antigens for the serodiagnosis of Fasciola gigantica infection in cattle, sheep and donkeys

An indirect enzyme-linked immunosorbent assay (iELISA) was evaluated for its diagnostic capability in detecting antibodies against Fasciola gigantica infection in cattle, sheep and donkeys sera using crude worm, excretory–secretory and glutathione S-transferase antigens prepared from adult liver fluke. Presence of F. gigantica worms at post-mortem examination of cattle, sheep and donkey’s livers was taken as a gold standard for the evaluation of the assay. The diagnostic sensitivity, specificity and accuracy percentages of iELISA were determined for each antigen. Excretory–secretory antigen gave the best results for the serodiagnosis of F. gigantica infection in cattle, sheep and donkeys using iELISA with diagnostic sensitivity percentages of 93.3%, 94.9% and 93.3%, respectively, while the specificity percentages were 96.7%, 97.2% and 96.3%, respectively, whereas the accuracy percentages were 95%, 96% and 95.7%, respectively. The diagnostic sensitivity percentages of iELISA using crude worm antigen were 96.7%, 100% and 93.3%, respectively, while the specificity percentages were 80%, 83.3% and 85.2%, respectively, whereas the accuracy percentages were 88.3%, 86.7% and 87%, respectively. The diagnostic sensitivity percentages of iELISA using glutathione S-transferase antigen were 66.7%, 71.8% and 60%, respectively, while the specificity percentages were 70%, 77.8% and 77.8%, respectively, whereas the accuracy percentages were 68.3%, 74.7% and 73.9%, respectively. Conclusively, excretory–secretory antigen dependent iELISA can be used as a reliable serodiagnostic test for F. gigantica infection in cattle, sheep and donkeys.     

Trends in antimicrobial susceptibility among isolates of Campylobacter species in Ireland and the emergence of resistance to ciprofloxacin

Measurements were made of the susceptibility to six commonly prescribed antibiotics, including erythromycin, tetracycline and ciprofloxacin, of 130 isolates of Campylobacterjejuni and 15 isolates of Campylobacter coli cultured from human and poultry sources during 2000. The results were compared with the results from a collection of strains isolated between 1996 and 1998. The levels of resistance to erythromycin remained low, 2 per cent and 4.4 per cent for the human and poultry isolates, respectively. Resistance to tetracycline had increased to 31 per cent and 24.4 per cent from 13.9 per cent and 18.8 per cent for the human and poultry isolates, respectively. However, the resistance to ciprofloxacin of the strains isolated during 2000 had increased to 30 per cent, whereas between 1996 and 1998 there had been no resistance to this agent among human isolates, and only 3.1 per cent resistance among poultry isolates. The molecular basis for this resistance has been shown to be the result of a single amino acid substitution, Thr-86-Ile, in the gyrA subunit of DNA gyrase in Cjejuni. A subset of 59 isolates was tested by molecular methods and all of the 25 phenotypically resistant isolates possessed this substitution. None of the human isolates had been treated with ciprofloxacin before their laboratory isolation.     

Studies on the efficacy of different adjuvants in live stock specific bacterial vaccines for turkeys against Bordetella infection and onset of antibody titers in respect to the age of the turkey poults

For evaluating the influence of the age of the vaccinated birds on the development of antibodies, five groups of turkey poults were inoculated subcutaneously at day 1, 7, 10, 14 and 21 of life with vaccine containing inactivated Bordetella avium and Freund's incomplete adjuvant. No matter which vaccine schedule was used, serum antibodies with the ELISA were first detected at the 28th day of life and increased continuously until the 49th day, when it exhibited either a peak or a plateau. Aluminium hydroxide, Freund's complete and incomplete adjuvant and a mineral oil-arlacel-tween-mixture being permitted adjuvants (appendix II EWG 2377/90) and the adjuvant Gerbu 100 were evaluated for their suitability. Turkeys were vaccinated at the age of three weeks and examined clinically as well as serologically up to the 11th week. Humoral antibodies were detected quantitatively using an ELISA for IgG and a microagglutination test for IgM and qualitatively using immunodiffusion. The damage at the application site was rated by measurement of the swelling of the tissue. In the 10th week, the animals were infected with the agent for challenge. The serological examination for IgG antibodies in the ELISA both treatments with Freund's adjuvants resulted in high titers, which differed significantly from the unvaccinated control after 21 days. IgM could be detected from day 7 onwards in all vaccinated groups and showed the highest titers when aluminium hydroxide was used as adjuvant. In the immunodiffusion assay, precipitating antibodies could be found from the first week after vaccination onwards. There was no correlation between the occurrence of precipitating antibodies and ELISA titers. The measurements of the swelling of the tissue in the beginning showed the largest swellings in the animals injected with Freund's incomplete adjuvant and differed significantly from the unvaccinated control. In the 10th week, the animals were infected with Bordetella avium for challenge. In comparison to the unvaccinated animals, all vaccinated turkeys, no matter which adjuvant was used, showed a distinct and significant reduction of the reisolation rate.     

Changes in the bacterial flora of the upper and lower respiratory tracts and bronchoalveolar lavage differential cell counts in feedlot calves treated for respiratory diseases.

Serial nasopharyngeal swab and bronchoalveolar lavage cultures were used to estimate changes in the bacterial flora of the respiratory tracts of calves during the first month after arrival in the feedlot. Bronchoalveolar lavage (BAL) differential cell counts served to evaluate pulmonary inflammatory changes during this period. Two groups of calves were studied, one consisting of clinically normal controls (n = 60), the other, of cases (n = 59) which received treatment for respiratory disease (penicillin +/- trimethoprimsulfadoxine). A variety of organisms, including Pasteurella multocida, Pasteurella haemolytica, Haemophilus somnus, Mycoplasma bovis and Mycoplasma bovirhinis, were present in the upper and lower airways of both groups during the postarrival period. With the exception of M. bovis, an overall decline in the prevalence of these organisms was observed during the course of the study. In cases, there was a marked decrease in the number of Pasteurella spp. and H. somnus isolates immediately following treatment. For the Pasteurella spp., however, this effect was shortlived as they often appeared to recolonize the respiratory tract within eight days of terminating antimicrobial therapy. Treatment did not appear to affect the frequency of isolating M. bovis. Its prevalence, in both groups of calves, increased to levels approaching 100% during the course of the study. All Pasteurella spp. isolates were tested for susceptibility to several commonly used antimicrobials. Resistance was only evident among P. haemolytica isolated from cases and in every instance this was to a combination of penicillin, ampicillin and tetracycline. Significantly more isolates were resistant after treatment than before. There were BAL differential cell count abnormalities indicative of inflammation in both cases and controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Diagnostic utility and cost‐effectiveness of reflex bacterial culture for the detection of urinary tract infection in dogs with low urine specific gravity

Background: Urinary tract infections (UTIs) may be subclinical or difficult to detect in dilute urine as sediment abnormalities may not be observed. In our laboratory, bacterial culture is automatically performed (reflex culture) on samples with urine specific gravity (USG)≤1.013 to increase the likelihood of detecting infection. The value of routine culture of dilute urine, however, has not been fully assessed. Objective: The purpose of this retrospective study was to evaluate the frequency of positive bacterial cultures and analyze the diagnostic utility and cost‐effectiveness of culture compared with routine sediment examination for detecting UTI in dilute urine specimens from dogs. Methods: Urinalysis and concurrent aerobic bacterial culture results were obtained from the electronic medical record system at the University of California–Davis Veterinary Medical Teaching Hospital for samples with USG≤1.013 analyzed from July 1998 through January 2005. Urine collection method, presence of leukocytes and bacteria, bacterial culture results, and clinical diagnosis were recorded. Cost‐effectiveness of reflex culture, based on low USG as the sole criterion, was evaluated. Results: Of 1264 urine specimens, 106 (8.4%) had positive bacterial cultures. Using culture as the gold standard, sediment evaluation had a diagnostic sensitivity of 58.5% and specificity of 98.3% (diagnostic accuracy 94.9%). An additional cost of $60 per patient was incurred, leading to average annual costs of $11,668 for reflex bacterial cultures of all samples with low USG, regardless of collection method. Within our study population, 10 urine samples needed to be cultured for each true positive result. Conclusions: The sensitivity of urine sediment evaluation is low for UTI in dilute urine samples; however, reflex bacterial culture does not appear to be cost‐effective in dogs with USG≤1.013 in the absence of active urine sediment or high clinical suspicion for UTI.     

上海市柑桔主要病虫害综合防控技术介绍

经过1998年至2011年观察研究比较,认为上海地区柑桔病虫害主要有六种,为了提供优质、安全的果品,在柑桔病虫害的防治方面宜采取农业防治为基础,物理生物防治为主,化学农药防治为辅的综合防治方法,现在分别对这六种最主要的病虫害进行发生规律和防治方法的介绍。     

Establishment of the PCR system specific to Salmonella spp. and its application for the inspection of food and fecal samples

We established the PCR detection system specific to Salmonella species using Salmonella enterotoxin gene (stn). The detection limit was one bacterial cell per one gram of fecal and minced-meat samples using enrichment procedure by Tripticase soy broth or Salmonella enrichment broth, respectively. We concluded that this PCR system is useful for the practical application in the field of the public hygiene.     

Estimation of receiver-operating characteristic curves to determine accuracy of a competitive enzyme-linked immunosorbent assay for the serodiagnosis of Brucella infection in domestic water buffalo (Bubalus bubalis) and cattle

OBJECTIVE: To estimate receiver-operating characteristic (ROC) curves for a competitive ELISA (c-ELISA) that is used in serodiagnosis of brucellosis in water buffalo and cattle, to determine the most appropriate positive cutoff value for the c-ELISA in confirmation of infection, and to evaluate species differences in c-ELISA function. SAMPLE POPULATION: Sera from 4 herds of cattle (n = 391) and 4 herds of water buffalo (381). PROCEDURE: Serum samples were evaluated for Brucella-specific antibodies by use of a c-ELISA. On the basis of previous serologic test results, iterative simulation modeling was used to classify animals as positive or negative for Brucella infection without the use of a gold standard. Accuracy of c-ELISA for diagnosis of infection was compared between cattle and water buffalo by comparison of areas under ROC curves. RESULTS: A positive cutoff value of 30% inhibition for c-ELISA yielded sensitivity and specificity estimates, respectively, of 83.9 and 92.6% for cattle and 91.4 and 95.4% for water buffalo. A positive cutoff value of 35% inhibition yielded sensitivity and specificity estimates, respectively, of 83.9 and 96.2% for cattle and 88.0 and 974% for water buffalo. Areas under ROC curves were 0.94 and 0.98 for cattle and water buffalo, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: ROC curves can be estimated by use of iterative simulation methods to determine optimal cutoff values for diagnostic tests with quantitative outcomes. A cutoff value of 35% inhibition for the c-ELISA was found to be most appropriate for confirmation of Brucella infection in cattle and water buffalo.     

青藏高原东部常见禾本科植物种子大小变异及其与萌发的关系

禾本科植物在青藏高原分布较广,选取该地47种常见禾本科植物,观测其种子大小变异,并在人工气候室内做萌发试验,研究了在变温(12 h 10℃/12 h 20℃)、无光照条件下,禾本科植物的种子大小对萌发特性的影响。结果表明:高寒草甸禾本科植物种子百粒重范围为0.006 9~1.436 3 g,平均重量为0.192 1 g,重量大小变异达到了近3个数量级。60%的种子在0.006 9~0.148 9 g,种子大小聚类分析后,归为组1的有28个物种(0.006 9~0.148 9 g),说明高寒草甸禾本科以较小的种子占绝对优势。种子大小与萌发指数、萌发率都成正相关,与萌发开始时间呈负相关,这说明对于高寒草甸禾本科植物来说,种子越小萌发越快,萌发开始时间越早,而萌发率却越低,反之亦然。试验结果反映了禾本科植物种子大小在一定程度上对萌发特性起着重要的作用。     

Correlation between the serum neutralization test and the indirect immunofluorescent test for the detection of specific antibodies to infectious bovine rhinotracheitis virus.

Comparative titrations show that immunofluorescence on infected rabbit kidney cells is relatively as sensitive as the neutralization test for the detection of antibodies to the infectious bovine rhinotracheitis virus. When calf testis cells infected with this virus are used for the detection of antibodies by the indirect immunofluorescent test nonspecific staining is high. This nonspecific reaction is absent when rabbit kidney cells are used.     

The transfer of maternal IgE and other immunoglobulins specific for Trichostrongylus colubriformis larval excretory/secretory product to the neonatal lamb

The transference of immunoglobulins from six New Zealand Romney ewes to their lambs was examined. Immunoglobulin levels were determined in ewe plasma, colostrum and lamb plasma shortly after birth and before the lambs fed, in lamb plasma 2 days after birth, and lamb plasma, ewe plasma and milk 30 days after parturition. Levels of total IgE, and IgE, IgG1, IgG2, IgM, and IgA with specificity for Trichostronglus colubriformis third stage larval secretory/excretory products (TcL3E/S) were determined. Mean levels of total IgE were three times higher in colostrum than in parturient ewe plasma while only trace amounts were detected in milk at 30 days after birth (107.7, 34.3, and 0.2U ml(-1), respectively, differences between means P< or =0.01). Mean total IgE in lamb plasma rose from being undetectable before suckling to levels comparable to those of the ewes by 2 days after birth (21.7U ml(-1)) and then declined to low levels by 30 days (0.4U ml(-1)). Total IgE levels in lamb plasma were significantly correlated with levels in ewe plasma and colostrum (r=0.91, P< or =0.01; r=0.96, P< or =0.003, respectively). The transference of TcL3E/S-specific IgE, IgG1 and IgA was substantial with mean levels of these antibodies in lamb plasma at 2 days comparable to that in parturient ewe plasma (absorbance levels in lamb plasma of 0.283, 0.537, and 0.334, respectively). Proportionally less maternal IgM and IgG2 appeared to be transferred to the lambs (absorbance of 0.112 and 0.081, respectively). Levels of TcL3E/S-specific IgE and IgG1 in lamb plasma at 2 days were significantly correlated with levels in parturient ewe plasma and colostrum (r=0.89 and 0.82, 0.85 and 0.96; all P< or =0.05, respectively). These results indicate that IgE is concentrated in ewe colostrum and that substantial amounts of maternal IgE are transferred to lambs via colostrum. Further, the results suggest that humoral immunity against gastro-intestinal nematode parasites and potentially other parasites in colostrum-fed lambs may approximate that of the ewe. The implications of the transference of humoral immunity through colostrum in ruminants for the passive protection and the development of active immunity against parasites remains to be fully explored.     

Pulsed-wave Doppler tissue imaging velocities in normal geriatric cats and geriatric cats with primary or systemic diseases linked to specific cardiomyopathies in humans, and the influence of age and heart rate upon these velocities

Pulsed-wave Doppler tissue imaging (pw-DTI) techniques allow the non-invasive assessment of myocardial dynamics. pw-DTI has demonstrated regional and global diastolic impairment in various forms of human and feline cardiomyopathy. We hypothesise that in geriatric cats with systemic diseases that have been linked to specific cardiomyopathies in human beings, the myocardial velocity profile will be altered when compared to either normal or hypertrophic cardiomyopathy (HCM) cats; and that both age and heart rate have a significant affect upon pw-DTI velocities. The aims of this study were to determine whether the feline M-mode or myocardial velocity profile is altered in geriatric cats with disease states that have been linked to specific cardiomyopathies in humans when compared to normal geriatric cats or geriatric cats with HCM and to determine whether age or heart rate has a significant effect upon pw-DTI velocities within these groups of cats. Sixty-six cats aged 8 years or above were included in the study, and were divided as follows: Unaffected (n=8), basilar septal bulge (BSB) (17), HCM (14), hyperthyroid (HiT(4)) (12) and chronic renal failure (CRF) (15). Systolic blood pressure was normal in all the cats. pw-DTI systolic (S'), early (E') and late diastolic (A') velocities were assessed from standardised sites within the myocardium, and the relationships between these and disease group, age and heart rate were then assessed. In cats with HCM, the E' velocity was decreased at various sites. Conversely, the HiT(4) cats demonstrated increased S' velocities. The only site at which the age of the cat was significantly related to myocardial velocities was the S' velocity from the apical mid-septum. There were also significant positive relationships between heart rate and the magnitude of myocardial S', E' and A' velocities of radial motion and S' and A' velocities of longitudinal motion. pw-DTI detected diastolic dysfunction in untreated cats with HCM and increased systolic function in HiT(4) cats. The age of the cat was of little significance, whereas heart rate significantly influenced myocardial velocity profiles.