Nasal lesions in rats exposed to hydrogen sulfide for four hours

Fischer-344 rats were exposed for 4 hours to 0, 14, 280, or 560 mg of hydrogen sulfide.m-3 and killed 1, 18, or 44 hours later. We evaluated the nasal epithelial cells and determined the anatomic distribution of lesions. Inhalation of 560 mg of hydrogen sulfide.m-3 induced necrosis and exfoliation of respiratory and olfactory mucosal cells, but not squamous epithelial cells. The anatomic distribution of lesions was midway along the nasal passages involving nasal and maxillary conchae, but not ethmoidal conchae. Injured respiratory mucosa repaired rapidly, whereas olfactory mucosa continued to exfoliate at 44 hours after exposure.     

Histologic and ultrastructural alterations in lungs of rats exposed to sub-lethal concentrations of hydrogen sulfide

Fischer-344 rats were killed 1, 18, and 42 hr after a single 4-hr exposure to an atmosphere of 0, 116, or 615 mg m-3 of hydrogen sulfide (H2S). Lungs, fixed by the intratracheal route, were examined by light and electron microscopy. Histologic changes were transient and mainly present in rats exposed to 615 mg m-3 H2S. Lesions included severe but transitory pulmonary edema and fibrinocellular alveolitis which was restricted to the proximal alveolar region of the lung. Electron microscopically, ciliated bronchiolar cells were the only cells that developed necrosis; they were rapidly replaced by mitosis. Alveolar endothelium had cytoplasmic blebs, but alveolar epithelium had minor changes. No mast cell degranulation was detected in lungs with edema. A 4-hr exposure to 615 mg m-3 is markedly edematogenic for the lung but only moderately cytotoxic for pulmonary cells.     

Effects of cyclophosphamide on the kaolin consumption (pica behavior) in five strains of adult male rats

It is known that pica, the consumption of non-nutritive substances such as kaolin, can be induced by administration of toxins or emetic agents in rats. In the present study, we examined the effects of intraperitoneal (i.p.) administration of cyclophosphamide on pica behavior and on the concentration of 5-hydroxyindoleacetic acids (5HIAA) in cerebrospinal fluid (CSF) in the following five strains of adult male rats: Sprague Dawley (SD), Wistar, Fischer 344 (F344), Wistar-Imamichi (WI) and Long Evans (LE). Cyclophosphamide (25 mg or 50 mg/kg) was injected (i.p.) into the rats and kaolin and food intake were measured at 24 hr after injection. The animals were anesthetized with urethane (1 g/kg) at 3 hr after injection of cyclophosphamide, and CSF was collected from the cisterna magna. WI and LE rats clearly showed pica behavior as compared with the other strains. In LE rats, the concentration of 5HIAA in CSF also increased in a dose-dependent manner of cyclophosphamide. The pretreatment with ondansetron (5-HT(3) antagonist) restored both changes (kaolin consumption and 5HIAA levels) induced by cyclophosphamide. These results suggest that the LE rat is sensitive to cyclophosphamide, that pica induced by cyclophosphamide mimics many aspects of emesis including the serotonergic response in the central nervous system and that use of the pica model would be a practical method for evaluating the effects of antiemetic drugs in addition to the mechanism of emesis.     

三种药物对西施舌稚贝的急性毒性研究

在常温静水条件下,进行敌百虫、甲醛和鳗菌净对西施舌稚贝的急性毒性试验。结果表明:敌百虫、甲醛和鳗菌净对西施舌稚贝的24hLC50分别为26.75mg/L、54.25mg/L、1.47mg/L,其安全浓度分别为0.27mg/L、0.54mg/L、0.01mg/L。本文探讨如何有效地使用药物杀灭西施舌稚贝的敌害生物,以保证西施舌人工育苗成功。     

稻草与不同水平百脉根组合的瘤胃体外发酵及甲烷产量的动态变化研究

为研究稻草(RS)添补百脉根(LC)的瘤胃体外发酵及甲烷产量的动态变化,本试验采用单因子试验设计,体外批次培养48h,研究RS分别补添0%、10%、20%、30%、40%、50%、60%、70%、80%、90%与100%的百脉根(LC0、LC10、LC20、LC30、LC40、LC50、LC60、LC70、LC80、LC90与LC100)在3h、6h、12h、24h、36h与48h的瘤胃体外发酵参数与甲烷产量。结果显示,3~48h各组的pH值在6.64～6.99,NH3-N浓度为2.98~28.74mg/100mL,甲烷产量为0.038~1.313mmol。24h前甲烷产量随LC比例的增加而增加,多数组间差异不显著(P>0.05),24~48h则随LC比例的增加而减少,组间差异显著(P<0.05)。表明反刍动物RS基础日粮与LC组合可以降低甲烷产量。     

Critical evaluation of the modified-adult immersion test with discriminating dose bioassay for Boophilus microplus using American and Australian isolates

Similar adult immersion tests (AITs) for acaricide susceptibility of Boophilus microplus were done in Texas, USA (Mu?oz strain) and in Queensland, Australia (N-strain and Ultimo isolates). Engorged adult female ticks were immersed in one of a series of dilutions of commercial acaricide in water and then incubated at room temperature for 7 days. Data on oviposition were collected 7 days after exposure to acaricide and subjected to probit analysis. For most data, we observed poor fit to the probit model. Substantial differences in both LC50 and LC99 for the susceptible strains occurred between the respective laboratories and confidence intervals for all acaricides and all strains were unacceptably wide. For amitraz, the discriminating concentration (double the LC99.9 or LC99) recommended by FAO was 0.25%, but our estimates ranged from 0.46% to 9000%. For cypermethrin, the recommended DD was 0.0050%, with our estimates ranging from 0.00022% to 0.74%. For coumaphos the recommended DD was 0.50% but our estimates ranged from 0.66% to 130%. Finally, for moxidectin, the recommended DD was 0.10%, while our estimates ranged from <0.0001% to 5.9%. The method does not provide a means to discriminate between amitraz-susceptible and -resistant, nor between cypermethrin-susceptible and -resistant B. microplus.     

Assessment of diclofenac LC50 reference values in juvenile and embryonic stages of the zebrafish (Danio rerio)

The aim of the study was to compare the acute toxicity of diclofenac to juvenile and embryonic stages of the zebrafish (Danio rerio). Acute toxicity tests were performed on the aquarium fish Danio rerio, which is one of the model organisms most commonly used in toxicity testing. The tests were performed using a semi-static method according to OECD guideline No. 203 (Fish, acute toxicity test). Embryo toxicity tests were performed in zebrafish embryos (Danio rerio) in compliance with OECD No. 212 methodology (Fish, short-term toxicity test on embryo and sac-fry stages). The results were subjected to a probit analysis using the EKO-TOX 5.2 programme to determine 96hLC50 and 144hLC50 (median lethal concentration, 50% mortality after a 96 h or 144 h interval, respectively) values of diclofenac. The statistical significance of the difference between LC50 values in juvenile and embryonic stages of Danio rerio was tested using the Mann-Whitney non-parametric test implemented in the Unistat 5.1 programme. The LC50 mean value of diclofenac was 166.6 +/- 9.8 mg/L in juvenile Danio rerio, and 6.11 +/- 2.48 mg/L in embryonic stages of Danio rerio. The study demonstrated a statistically higher sensitivity to diclofenac (P < 0.05) in embryonic stages compared to the juvenile fish.     

3种半数致死浓度计算方法之比较

通过对寇氏法、概率单位法及线性回归法等3种半数致死浓度(50%lethal concentration,LC50)计算方法的阐述,比较了其基本公式与操作方法等。结果表明不同时间的半数致死浓度均依线性回归法、寇氏法、概率单位法的顺序逐步升高,但差异性不显著(P>0.05)。在操作上,概率单位法和线性回归较寇氏法更容易掌握。     

Syringe test (modified larval immersion test): a new bioassay for testing acaricidal activity of plant extracts against Rhipicephalus microplus

We report a new bioassay "syringe test" (modified larval immersion test) for in vitro evaluation of acaricidal activity of crude plant extracts. Prepared syringes, containing eggs of tick, were incubated until 14 d after hatching of eggs, when the bioassay was performed on the larvae. Lethal concentrations for 50% of larvae (LC(50)), LC(90) and LC(99) values were calculated for each tested product. 95% confidence intervals for LC(50) were very narrow, indicating a high degree of repeatability for the new bioassay on larvae of R. microplus. Bioassays were applied to six crude aqueous-methanol extracts from five plants (Acacia nilotica, Buxus papillosa, Fumaria parviflora, Juniperus excelsa, and Operculina turpethum), of which three showed discernible effects. Twenty-four hours post exposure, LC(99) values were 11.9% (w/v) for F. parviflora, 20.8% (w/v) and 29.2% (w/v) for B. papillosa and A. nilotica, respectively. After six days of exposure these values were; 9.1% (w/v), 9.2% (w/v) and 15.5 (w/v) for F. parviflora, A. nilotica and B. papillosa, respectively.     

唐古特莨菪中生物碱的分离、鉴定与生物活性研究

研究唐古特莨菪中生物碱对农业害虫(螨)的生物活性,可为利用该植物研制新型植物源杀虫(螨)剂提供理论依据。本文采用柱色谱分离法从唐古特莨菪地上部分分离得到2个生物碱类化合物,通过MS、NMR确定了其化学结构,分别为莨菪碱、东莨菪碱。采用微量点滴法和玻片浸渍法测定了莨菪碱、东莨菪碱对甘蓝蚜、桃蚜等9种蚜虫和朱砂叶螨、二斑叶螨等4种叶螨的触杀活性。结果表明,莨菪碱对禾谷缢管蚜、桃粉蚜、豆蚜和绣线菊蚜具有较强的触杀作用,LC₅₀值分别为257.863,275.459,344.645和344.717 mg/L。东莨菪碱对豆蚜和禾谷缢管蚜具有较强的触杀作用,LC₅₀值分别为311.585和392.309 mg/L。对照药剂鱼藤酮对甘蓝蚜和棉蚜具有较强的触杀作用,LC₅₀值分别为399.542和436.124 mg/L。对甘蓝蚜等9种蚜虫的触杀作用,莨菪碱强于或与东莨菪碱相当;对豆蚜、禾谷缢管蚜的触杀作用,莨菪碱和东莨菪碱均强于鱼藤酮;对甘蓝蚜、桃蚜和棉蚜的触杀作用,莨菪碱和东莨菪碱均弱于鱼藤酮。莨菪碱和东莨菪碱对截形叶螨、山楂叶螨、二斑叶螨和朱砂叶螨具有较强的触杀作用。2种生物碱对4种叶螨的毒力相当,对二斑叶螨和朱砂叶螨的毒力(201.027~224.172 mg/L)均高于截形叶螨和山楂叶螨(257.014~332.698 mg/L)。对照药剂鱼藤酮对截形叶螨、山楂叶螨的LC₅₀值分别为196.847和224.592 mg/L,其毒力高于莨菪碱和东莨菪碱。     

Pharmacokinetics of a single intravenous dose of marbofloxacin in adult donkeys

Six donkeys each received 2 mg/kg marbofloxacin as a 10 per cent aqueous solution administered intravenously. Principal pharmacokinetic parameters were determined and two efficacy indices were computed by using pharmacokinetic parameters and selected mic90 values of marbofloxacin against pathogenic equine strains to predict the efficacy of the drug at this dose. The pharmacokinetics of marbofloxacin in donkeys was characterised by a large mean volume of distribution at a steady state (1.15 [0.09] l/kg) and a long mean (sd) elimination half-life of 9.24 (1.96) hours. It was also characterised by a relatively slow total body clearance of 0.10 (0.02) l/kg/hour, slower than in horses. Using mic90 values of marbofloxacin against pathogenic equine strains with a daily dose of 2 mg/kg, appropriate values of efficacy indicators were obtained only for Enterobacteriaceae. Daily intravenous doses of 0.33, 2.62 and 20 mg/kg were calculated for evaluation in clinical trials of infections due to Enterobacteriaceae, Staphylococcus aureus and Streptococci, respectively.     

结肠小袋纤毛虫体外药物急性毒杀试验

通过结肠小袋纤毛虫对甲硝唑、氟苯尼考和血虫净（三氮咪）的急性毒性试验,以机率单位法获得半数致死浓度（LC50）,结果显示：甲硝唑、氟苯尼考对结肠小袋纤毛虫2h的半数致死浓度（LC50,2h）分别是457．8mg／L,1705mg／L,12h的半数致死浓度（LC50,12h）分别是25．64mg／L、973．8mg／L。血虫净（三氮咪）12h和24h半数致死浓度（LC50）分别是331．8mg／L、295．4mg／L。甲硝唑是杀灭结肠小袋纤毛虫的理想药物。     

Toxicity of cypermethrin and diazinon to Haematobia irritans (Diptera: Muscidae) in its American southern range

Synthetic pyrethroids are considered currently less effective than organophosphate insecticides for controlling Haematobia irritans on cattle in its American southern range. To test the validity of this hypothesis, horn fly populations from 95 cattle herds in central-northern Argentina and southern Brazil were evaluated to determine 50% lethal concentrations (LC(50)) to cypermethrin and diazinon, respectively. Horn flies were exposed to filter papers and glass vials treated with cypermethrin and diazinon, respectively, and LC(50) values were determined using probit analysis. Two and 78 fly populations showed LC(50) values indicating susceptibility and resistance to cypermethrin, respectively, in comparison to historical LC(50) values obtained when the horn fly populations were susceptible to cypermethrin. Five fly populations had an undetermined resistance status to cypermethrin. The diazinon tests showed no obvious indications of H. irritans resistance. Thirty nine percent of farmers still rely on pyrethroids to control horn fly populations on cattle which represents a decreasing proportion from a few years ago when horn fly control was carried out using pyrethroids almost exclusively. Organophosphates could provide control for horn flies resistant to pyrethroids; however, the reliance on chemicals alone to control H. irritans is not considered a proper approach for the future.     

Determination of the susceptibility of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) to ivermectin and fipronil by Larval Immersion Test (LIT) in Uruguay

Rhipicephalus (Boophilus) microplus is an important cattle pest in Uruguay, and the law regulates its control. It is resistant to organophosphates, synthetic pyrethroids and, as recently discovered, to fipronil. Resistance to macrocyclic lactones (MLs) and amitraz have not been documented; however, veterinarians and farmers have reported treatment failures. The objective of the present work was to study the susceptibility of cattle tick strains from different Uruguayan counties to ivermectin (IVM) and fipronil by using the Larval Immersion Test (LIT). The Mozo strain was used as the susceptible reference strain. From 2007 to 2009, twenty-eight tick populations were collected from different cattle farms with and without history of IVM or fipronil use. A probit analysis estimated dose-mortality regressions, lethal concentrations (LC), and confidence intervals. The resistance ratio (RR) was determined at the LC(50) and LC(90) estimates. To classify a tick population in relation to resistance, three categories based on a statistical analysis of LC and RR between field populations and Mozo strains were defined: susceptible (no differences), incipient resistance (differences and RR(50)<2) and resistant (differences and RR(50)≥2). Eighteen field populations were tested with IVM and five of them presented a RR(50) range between 1.35 and 1.98 and the LC(50/90), which is statistically different from the Mozo strain (incipient resistance). However, the RR(90) increases ≥2 in four of the populations, confirming that tick resistance to IVM is emergent. The low RR values obtained could be a result of a low frequency of treatments with IVM. Twenty-seven tick populations were tested with fipronil and six were diagnosed as resistant according to the LIT. Cross-resistance was not observed between fipronil and IVM on these tick populations. The current study presents different R. (B.) microplus populations with an incipient resistance to IVM, and indicates that the fipronil tick resistance is restricted to certain areas in Uruguay.     

Evaluation by scoring and computerized morphometry of lesions of early Mycoplasma pulmonis infection and ammonia exposure in F344/N rats

To evaluate lesions of ammonia exposure and Mycoplasma pulmonis infection in gnotobiotic F344/N rats, groups of rats were inoculated with M. pulmonis, exposed to 76 micrograms/liter (100 parts per million) ammonia, or both, or served as controls. Six rats from each group were killed 3, 5, 7, and 9 days after inoculation and their respiratory organs prepared for histologic examination. Lesions were assessed by subjective scoring and computerized morphometry, and results obtained by the two methods were compared to assess agreement of results. Lesions were limited to the nasal passages. Ammonia exposure resulted in hyperplastic and degenerative changes in the anterior nasal epithelium. Lesions of mycoplasmosis were more severe in ammonia-exposed than unexposed rats, but qualitative differences in lesions were not apparent. Results of the two methods agreed in assessment of submucosal inflammatory cell accumulation. Epithelial lesions were difficult to evaluate by both methods in infected, ammonia-exposed rats. In evaluation of lesions of ammonia exposure alone, agreement was adequate. Both methods indicated that exudates were significantly greater in ammonia-exposed infected rats than in non-exposed infected rats. Thus, scoring and morphometry provided similar assessments of differences in lesion severity among the treatment groups.     

Acetylsalicylic acid and blood coagulation in the horse.

Equine blood may contain salicylic acid (SA) taken up as free acid or represents the metabolite of acetylsalicylic acid (ASA). To obtain information of SA in race horses we screened blood samples of trotting-horses routinely drawn to be analyzed for doping substances. The individual values determined followed a Gaussian distribution displaying a geometric mean of 19 ng SA per ml serum. A probit analysis revealed linear relationship (r = 0.995). Additional studies examined the antithrombotic efficacy of ASA in the horse. An oral dose of 300 mg ASA considerably elevated the bleeding time for more than 2 hours with concomitant SA serum levels between 800 and 1000 ng/ml. It is concluded that salicylate levels even below 1 microgram/ml serum bring about considerable pharmacologic effects such as prolongation of bleeding time, decrease in blood viscosity and possibly dilatation of blood vessels. These effects may improve the tissue supply with blood including oxygen.     

Median infectious dose (ID₅₀) of porcine reproductive and respiratory syndrome virus isolate MN-184 via aerosol exposure

The median infectious dose (ID(50)) of porcine reproductive and respiratory syndrome (PRRS) virus isolate MN-184 was determined for aerosol exposure. In 7 replicates, 3-week-old pigs (n=58) respired 10l of airborne PRRS virus from a dynamic aerosol toroid (DAT) maintained at -4°C. Thereafter, pigs were housed in isolation and monitored for evidence of infection. Infection occurred at virus concentrations too low to quantify by microinfectivity assays. Therefore, exposure dose was determined using two indirect methods ("calculated" and "theoretical"). "Calculated" virus dose was derived from the concentration of rhodamine B monitored over the exposure sequence. "Theoretical" virus dose was based on the continuous stirred-tank reactor model. The ID(50) estimate was modeled on the proportion of pigs that became infected using the probit and logit link functions for both "calculated" and "theoretical" exposure doses. Based on "calculated" doses, the probit and logit ID(50) estimates were 1 × 10(-0.13)TCID(50) and 1 × 10(-0.14)TCID(50), respectively. Based on "theoretical" doses, the probit and logit ID(50) were 1 × 10(0.26)TCID(50) and 1 × 10(0.24)TCID(50), respectively. For each point estimate, the 95% confidence interval included the other three point estimates. The results indicated that MN-184 was far more infectious than PRRS virus isolate VR-2332, the only other PRRS virus isolate for which ID(50) has been estimated for airborne exposure. Since aerosol ID(50) estimates are available for only these two isolates, it is uncertain whether one or both of these isolates represent the normal range of PRRS virus infectivity by this route.     

Experimental rapid infection in market swine following exposure to a Salmonella contaminated environment

The objective of these experiments was to evaluate the possibility of swine becoming infected with Salmonella Typhimurium after a short time interval in a contaminated environment. Two experiments were conducted. Experiment 1 consisted of five trials with eight market weight swine. Pigs were necropsied at 2 (n = 10), 3 (n = 10) and 6 (n = 5) hours after continuous exposure to an environment contaminated with feces shed by swine intranasally inoculated with nalidixic acid-resistant Salmonella Typhimurium (chi 4232). In Experiment 2, pigs were necropsied after 30 minutes (n = 6), 60 minutes (n = 6), 2 hours (n = 6), and 6 hours (n = 3). In addition, control animals with no exposure were also necropsied in both experiments. At necropsy, the superficial inguinal, ileocecal, and mandibular lymph nodes, as well as cecal contents, distal ileum portion, and feces were evaluated. All samples were cultured for the presence of the nalidixic acid-resistant Salmonella. Feces deposited on the floor by intranasally inoculated swine were mixed with water to form slurry with a resulting load of 10(3)-10(5) Salmonella Typhimurium CFU per gram. In Experiment 1, 80% percent of animals with a 2-hour, 60% of animals with a 3-hour, and 100% of animals with a 6-hour exposure to this slurry had at least one sample test positive for the marked Salmonella Typhimurium strain. In Experiment 2, 50% of the 30 minute, 50% of the 60 minute, and 33% of the 2-hour exposed pigs had at least one sample test positive. These experiments show that market swine can become infected during routine resting or holding periods when exposed to relatively low levels (10(3) CFU) of Salmonella in the simulated pre-slaughter environment, and that exposure times as short as 30 minutes are sufficient to produce contaminated gastrointestinal tracts. They also demonstrate the high risk of holding pigs longer than six hours. Intervention at this step in the swine production process may have a significant impact on the safety of pork products.     

Estimation Models for the Morbidity of the Horses Infected with Equine Influenza Virus

Estimation formulas for the morbidity of horses infected with equine influenza virus by linear regression, logistic regression and probit transformation were developed, using data from the outbreak at the Sha Tin Racing Track in Hong Kong in 1992. Using these formulas, we estimated the equine influenza virus morbidity rates at training centers belonging to the Japan Racing Association (JRA) in October 1997 and in October 1998. In 1998 JRA started a new vaccination program, and every horse must now be vaccinated twice per year. At that time, the vaccine included two US lineage virus strains, the A/equine/Kentucky/81 strain and the A/equine/La Plata/93 (LP93) strain, against equine type-2 influenza viruses; it did not include any EU lineage virus strains, such as A/equine/Suffolk/89 (SF89). Comparing the geometric mean (GM) values of hemagglutination inhibition (HI) titers between the LP93 strain and the SF89 strain in 1997 and in 1998, they both rose significantly at every age (p<0.05) by Wilcoxon test. Calculations by the simulation models show the morbidity rates for LP93 diminished from 0.439 (linear), 0.423 (logistic) and 0.431 (probit) to 0.276 (linear), 0.265 (logistic) and 0.271 (probit), respectively. On the other hand, the estimated morbidity rates for SF89 diminished only slightly from 0.954 (linear), 0.932 (logistic) and 0.944 (probit) to 0.946 (linear), 0.914 (logistic) and 0.927 (probit), respectively. Our simulation models could estimate the effect of the vaccine on each of the equine virus strains represented by the morbidity of infected horses. Thus, they are useful for vaccine evaluation.     

Susceptibility of two-week old Lymnaea natalensis to some plant extracts

The molluscacidal potency of 17 Nigerian plants extracted by the unevaporated crude water (UECW) method was evaluated on two-week old Lymnaea natalensis Krauss. Five extracts were not active but extracts of Balanites aegytiaca, Blighia sapida, Boswellia dalzielii, Cissampelos mucronata, Detarium microcarpum, Kigelia africana, Opilia celtidifolia, Parkia clappertoniana, Polygonum limbatum, Pseudocedrela kotschyi, Nauclea latifolia and Securidaca longipedunculata were molluscacidal. There is potential for their future use in the integrated control of Lymnaea natalensis, as well as other snails. Mortality data for lethal concentration values for all extracts were analysed by use of probit transformation. The upper and lower fiducial limits of the LC50 (P = 0.05) were also determined.