Vaccination experiments in the gadoid haddock, Melanogrammus aeglefinus L., against the bacterial pathogen Vibrio anguillarum

Vibrio anguillarum is one of the primary pathogens responsible for high levels of fish mortality in the aquaculture industry, and among gadoids O2a and b are the most common pathogenic serotypes. In this paper a variety of studies were performed to assess the optimal route by which to challenge haddock against this pathogen, and an optimal regime to vaccinate haddock. The most efficient method to challenge haddock with V. anguillarum in this study was immersion in a bath containing 10(7)cfu/ml, where 60% mortality was seen. Subsequent experiments showed that juvenile haddock could be protected against bacterial challenge with V. anguillarum, with a significant reduction in mortalities observed amongst the vaccination treatments when compared to the unvaccinated controls. However, as seen previously in cod studies, vaccination did not induce a specific antibody response.     

Infection level of monogenean gill parasite, Diplozoon kashmirensis (Monogenea, Polyopisthocotylea) in the Crucian Carp, Carassius carassius from lake ecosystems of an altered water quality: What factors do have an impact on the Diplozoon infection?

Monogeneans are of great interest to the ecologists because of their simple life cycle and are considered as one of the important and sensitive parasites to any changes in water quality. The monogenean gill parasite, Diplozoon kashmirensis of the Carassius carassius was examined with the aim to evaluate the infection level and the factors influencing the infection. Our results showed that highest prevalence (34.22%) of D. kashmirensis was in the lake having high trophic status and least prevalence (10.90%) in the lake having least trophic status. Infection levels were significantly higher at the basins/sites having deteriorated water quality in comparison to the basins/sites having better water quality. The combined effect of pollutants and eutrophication showed decrease in intensity of D. kashmirensis in one of the polluted/hypertrophied site in the hypertrophic lake (Anchar Lake), while as synergistic effect (i.e. increase) on prevalence and intensity was observed in one of the polluted/hypertrophied site in the eutrophic lake (Dal Lake). Sex was not an important factor influencing the D. kashmirensis in three lakes. The study showed that condition factor was significantly higher in the hypertrophic lake and in an uninfected fish. In addition microhabitat influenced the Diplozoon infection. Most preferable site for the D. kashmirensis was middle sector of the gill. Furthermore, the gill parasite showed significantly positive correlation (r(p)=0.6, P<0.05) with water temperature in the hypertrophic lake, thus showing the impact of water temperature on this parasite. It was concluded that intra lake differences, season, condition factor, microhabitat and water temperature influenced the infection of D. kashmirensis. It seems from the data that eutrophic and hypertrophic habitats were favourable for the D. kashmirensis. The D. kashmirensis showed both antagonistic and synergistic response to the combined effect of pollution and eutrophication.     

Effect of aflatoxin B1 on resistance induced by Bordetella bronchiseptica vaccine in rabbits

The effects of aflatoxin B1 on the development of the immune response to oil-adjuvanted Bordetella bronchiseptica vaccine and on acquired resistance to bacterial challenge were studied in rabbits. The doses of aflatoxin used were insufficient to produce clinical intoxication. Rabbits were randomly assigned to three groups, each having six animals: control (T), vaccinated (V), and vaccinated plus aflatoxin (VA) at 0.05 mg/kg daily per os. Groups V and VA were vaccinated twice, and the three groups were subsequently challenged with virulent B. bronchiseptica. The average weight gain at weekly intervals was significantly reduced in group VA, and no statistically significant differences were found in the titers of agglutinating antibodies between groups V and VA. There were significant differences between groups V and VA in the extent and severity of the pneumonic process, group VA being most affected. Results indicated that agglutinating antibody titers were not related to the level of protection in the latter group. Other mechanisms, such as alveolar macrophage activity and cell-mediated immunity, were implicated in the impairment of the acquired resistance in rabbits subclinically intoxicated with aflatoxin.     

Evidence of aberration of the natural cytotoxic cell activity in Fundulus heteroclitus (Pisces: Cyprinodontidae) from the Elizabeth River, Virginia

There is a general agreement that exposure to high concentrations of polynuclear aromatic hydrocarbons (PAH) in sediments is associated with high frequencies of neoplasms in feral fish species. Since PAH modulate the activity of murine and amphibian natural cytotoxic (killer) cells, a leukocyte subpopulation that is believed to play an important role in immunosurveillance, we wished to determine if fish exposed to PAH could have an altered natural cytotoxic cell (NCC) activity. In the present study, mummichog (Fundulus heteroclitus L.) were collected from two sites in the Elizabeth River, VA that are heavily contaminated with PAH, and from a relatively unpolluted reference site in the York River, VA. The cytotoxic activity of anterior kidney and splenic leukocytes was tested against the tumor cell line K562. The leukocytes from Elizabeth River fish displayed a significant depression of the in vitro tumorilytic activity as compared with leukocytes from the York River fish. Analysis of leukocyte-tumor cell conjugates indicated that Elizabeth River fish leukocytes were unable to recognize and subsequently bind to the tumor target cells. This suggests an aberration in the early events of the cytotoxic mechanism. By keeping the fish in cleaner York River water for up to 28 weeks the suppressed NCC activity was reversed totally in one site, which is slightly contaminated, and partially in the other site, which is heavily polluted with creosote from an operating wood treatment plant. This indicates that the decreased NCC activity was related, at least in part, to exposure to the chemical pollutants in the Elizabeth River sediments.     

Phagocytic activity of macrophages of rainbow trout against Vibrio anguillarum and the opsonising effect of antibody and complement

The phagocytosis of Vibrio anguillarum by peritoneal macrophages from normal rainbow trout was enhanced by antibody and complement. Treatment of either macrophages or the bacteria by antibody also enhanced opsonisation. Five weeks after immunisation with V anguillarum, the phagocytic activity of macrophages from rainbow trout was increased significantly compared with the activity of those from unvaccinated fish. Although agglutinin titres did not increase until three weeks after immunisation, seven out of 10 fish challenged one week after immunisation survived, indicating that immunised fish had developed resistance to vibrio infection before significant levels of antibody or phagocytic activity became detectable.     

VA菌根对白三叶吸收水分和养分的影响

采用土培试验,研究水分胁迫条件下,施磷肥接种VA菌根菌对白三叶的生长,养分吸收和水分利用的影响.结果表明,土壤干旱严重则抑制地上部和根系的生长,影响植株吸收养分,降低水分利用率.随着施磷量的提高,该状况则得以缓解.在水分胁迫和正常供水条件下,施磷量相同,接种菌根组的地上和根系生物量干重显著高于不接种组,干物质累积增加,减轻水分胁迫对植株生长的抑制程度,改善了植株体内磷素养分状况,提高了白三叶对土壤磷的有效利用及水分利用率.     

Effects of altitude and variety on nutrient levels and epiphytes of oats北大核心CSCD

Oat crops are a major animal feed source in the Qinghai-Tibetan plateau and nearby areas. Typically，the crops are used for making silage rather than hay because of frequent rain at harvest in autumn. However，there is little data on how the yield，forage quality and levels of epiphytes that may affect silage quality of different oat varieties are affected by altitude. Therefore，four oat varieties（Longyan No. 3，Longyan No. 5，Baiyan No. 2 and Bayou No. 3） were planted in 8 localities forming an altitude gradient： Huangzhong （2295 m），Tianzhu （2797 m），Shandan （2860 m），Hezuo （2957 m），Haiyan （3052 m），Maqu （3474 m），Maqin （3765 m）and Chengduo （4217 m）. Measurements included crop yield and dry matter （DM），water soluble carbohydrate （WSC），crude protein （CP），neutral detergent fiber （NDF）and acid detergent fiber （ADF）contents，and counts of lactic acid bacteria （LAB），molds，yeasts and aerobic bacteria. It was found that there were significant altitude and variety effects on fresh yield，nutritional quality and microbial epiphyte counts. With increase in altitude，fresh yield，WSC，NDF and LAB counts increased（P<0.05），as much as 284. 00%，15. 49%，10.81% and 11.60% increment were observed at Chengduo site than that at Huangzhong，respectively；meanwhile DM，CP and yeast and mold counts were reduced by 15. 67%，36.27%，23.53% and 7.75% at Chengduo compared with Huangzhong site，respectively. Among the four tested varieties，Longyan No. 5 had the highest fresh yield （75605 kg·ha−1），WSC （201.4 g·kg-1 DM） and NDF （604. 2 g·kg-1 DM） at the Chengduo site，followed by Longyan No. 3. The highest CP （119.7 g· kg-1 DM） and mold counts （4.12 lg cfu·g-1 FM） were observed in Baiyan No. 2 at the Huangzhong site. Bayou No. 3 produced the highest LAB and yeast counts at Maqin and Chengduo sites. In general，Longyan No. 3 and Longyan No. 5 gave better performance at sites above 3000 m altitude；for sites below 3000 m，the four tested varieties were all found to be suitable for silage production. © 2022 Editorial Office of Acta Prataculturae Sinica. All rights reserved.     

Microbial contamination in human and animal drinking water

Water plays an important roll in the epidemiology of a lot of infectious diseases. Groundwater as well as surface water contains generally microorganisms of several species, which cannot always differentiated properly in autochthonous flora and contaminants with health significance. Sources of bacterial or viral contaminants may be feces from man and/or animals but with different counts in ground and surface water. With respect to water used for supplying farm animals it is stated that it must have initially drinking water quality. Since it generally looses this quality in this supply system the following requirements shall be met: Free of Salmonellas and/or Campylobacter in 100 ml, no E. coli in 10 ml, total bacterial count at 37 degrees C less than 1000 cfu/ml and total bacterial count at 20 degrees C less than 10,000 cfu/ml.     

多抗性保加利亚乳杆菌的诱变和筛选

通过紫外诱变和硫酸二乙酯诱变,筛选出抗生素抗性(头孢氨苄和氨苄西林)的保加利亚乳杆菌,同时模拟人体胃酸环境(pH1.5～4.5)和十二指肠高胆酸盐环境(0.1g/100mL～0.4g/100mL)对诱变后的保加利亚乳杆菌的抗性进行了研究。结果表明,保加利亚乳杆菌突变株可抗头孢氨苄和氨苄西林(均为2mg/mL),并且在pH2.5～4.5时具有较强的生存能力,培养4h活菌数仍达108cfu/mL以上,在pH1.5条件下仍有部分存活。在0.1g/100mL～0.3g/100mL胆酸盐条件下培养4h,活菌数仍达106cfu/mL以上,且能在0.4g/100mL胆汁盐中存活。     

Determination of the number of somatic cells in milk using the rapid diphenylamine DNA filter method

The rapid reaction of the diphenylamine agent with DNA was used for the determination of the counts of somatic cells in cow's milk, using the DNA filter method. The method is based on the filtration of a warmed (65-70 degrees C) mixture of milk with Triton X-100 through the Synpor nitrocellulose membrane filter, pore size 2 to 5 microns, and subsequent DNA determination of the collected somatic cells by the colour reaction of diphenylamine. A 2ml quantity of distilled water and 4 ml of diphenylamine reagent were added to the membrane filters with somatic cells. The mixture is warmed in water bath at 90 to 100 degrees C for 20 min., then it is cooled, centrifuged (3500 X g, 15 min.), and the optical density is measured at 595 nm. The relation 8 micrograms = 1 million cells was used for the conversion of DNA content to the counts of cells. The average variation coefficient of the determination was 5.9% and the coefficient of correlation between the diphenylamine DNA filter method and the direct microscopy of the somatic cells on membrane filters was r = 0.997. Using the diphenylamine DNA filter method, the counts of somatic cells can also be determined from milk samples stored in frozen condition or from the filters with collected cells kept at the temperature of 4 degrees C (10 days) or 25 degrees C (3 days). Milk stabilized with formaldehyde can also be used for the determination if stored at 4 degrees C.     

Survival of Listeria monocytogenes in wilted and additive-treated grass silage

Grass was field-dried to 3 different dry matter (DM) levels (200, 430 and 540 g/kg) and inoculated with 10(6)-10(7) cfu/g of a Listeria monocytogenes strain sharing a phagovar occasionally involved in foodborne outbreaks of listeriosis. Formic acid (3 ml/kg) or lactic acid bacteria (8 x 10(5)/g) with cellulolytic enzymes were applied only to forages with low and intermediate DM levels. Forages were ensiled in laboratory silos (1700 ml) and were stored at 25 degrees C for 30 or 90 days. After 90 days of storage, L. monocytogenes could not be detected in any silo, except one with the high dry matter grass without additive. After 30 days of storage, between 10(2) and 10(6) cfu L. monocytogenes/g silage were isolated from the untreated silages. Increasing the DM content from 200 to 540 g/kg did not reduce listeria counts possibly because of the lower production of fermentation acids (higher pH). In silages treated with additives, counts of L. monocytogenes were always lower than in silages without additive. In wet silages (DM 200 g/kg) both additives were effective, but in the wilted silages (DM 430 g/kg) only the bacterial additive reduced listeria counts below detection level. Listeria counts were highly correlated to silage pH (r = 0.92), the concentration of lactic acid (r = -0.80) and the pooled amount of undissociated acids (r = -0.83).     

Immersion booster vaccination effect on sea bass (Dicentrarchus labrax L.) juveniles

In each challenge 30 sea bass juveniles (mean weight 3.3 +/- 0.2 g SD) were used. During the whole experiment (water T: 18 +/- 1 degrees C) the fish were held in four 50l seawater independent recirculation systems (one fish group per 50l system). The protection to the pathogen Vibrio anguillarum was tested on booster vaccinated sea bass (Dicentrarchus labrax L.) juveniles. The vaccination was performed by immersion for 60 s in a commercial anti-V. anguillarum vaccine suspension. Booster vaccination took place 60 days after the initial immunization. Thirty days after the booster vaccination all the fish received intraperitoneally (IP) 3.0 x 10(6) cfu/fish (colony forming units) virulent V. anguillarum bacteria. The booster vaccination showed a strong protection effect on the challenged sea bass. In the next 20 days after the challenge the mortality was 0% among the booster vaccinated sea bass, 10% among the once vaccinated fish and 50% in the control group (unvaccinated fish). No mortality was observed among the unvaccinated sea bass injected IP with sterile normal saline by the challenge.     

Experimental infection by Vibrio anguillarum in mice and guinea pigs.

The fish pathogen Vibrio anguillarum causes a lethal infection in farmed fish characterized by hemorrhagic septicemia. There are no reports of experimental laboratory infections in warm-blooded animals. We investigated the effects of an intraperitoneal infection with different doses of a V. anguillarum suspension in mice and guinea pigs. The infection caused a 95-100% of mortality in 24-48 h. Hemorrhagic septicemia was observed at necropsy and confirmed by histological and hematological examination. Immunohistochemically positive bacterial clumps were detected exclusively in vessel lumen in all examined organs, including brain, and V. anguillarum was reisolated in pure culture from all organs, particularly from the kidney. Blood analysis showed erythropenia and leukopenia with granulocytosis in mice, platelet reduction and leukopenia with lymphocytosis in guinea pigs.     

益生菌干酪乳杆菌Zhang和双歧杆菌V9发酵豆乳的研究

对益生菌Lactobacillus casei Zhang和Bifidobacterium animalis V9单独及混合(1:1)发酵豆乳及以其制作的活性豆乳饮料进行研究,并测定了豆乳37℃发酵期间及活性豆乳饮料4℃28d贮藏期间的pH、TA、FAN和活菌数。结果表明,豆乳发酵过程中,混合发酵豆乳酸度变化大于单独发酵且可有效缩短发酵时间;B.animalis V9发酵的豆乳FAN含量变化最大。活性豆乳饮料贮藏期间,含有B.animalis V9活性豆乳饮料的pH值和TA变化不明显,含有L.casei Zhang活性豆乳饮料的pH值显著降低,TA显著升高;所有样品的FAN在贮藏7d期间显著下降,之后变化不明显;在28d贮藏结束时,L.casei Zhang和B.animalis V9单独及混合发酵豆乳饮料时活菌数分别是2.28×109cfu/g、4.27×108cfu/g、1.56×109cfu/g和2.8×108cfu/g。本研究初步表明,在豆乳中L.casei Zhang和B.animalis V9混合发酵优于单独发酵,在活性豆乳饮料中L.casei Zhang和B.animalis V9具有优良的贮藏稳定性,显示了其应用于发酵豆乳制品具有良好的潜力。     

A study to evaluate the field efficacy of ivermectin, fenbendazole and pyrantel pamoate, with preliminary observations on the efficacy of doramectin, as anthelmintics in horses

The efficacy of ivermectin, fenbendazole, pyrantel pamoate and doramectin was evaluated under field conditions at 2 sites in the Free State Province of South Africa. The study involved 25 horses at each site, divided into 5 groups of equal size. Ivermectin, fenbendazole and pyrantel pamoate were administered orally at doses of 0.2, 10 and 19 mg/kg respectively. Doramectin was administered by intramuscular injection at a dose of 0.2 mg/kg. Treatment efficacy was based on the mean faecal egg count reduction 14 days post treatment. At site A a faecal egg count reduction of 100% was found after treatment with ivermectin, fenbendazole and doramectin. A 96.1% reduction was found after treatment with pyrantel pamoate. At site B ivermectin and doramectin produced a 100% reduction in faecal egg counts, fenbendazole produced an 80.8% reduction and pyrantel pamoate a 94.1% reduction. Doramectin produced a 100% reduction in faecal egg counts at both sites, despite not being registered for use in horses. In addition, the results indicated reduced efficacy of fenbendazole at site B, which suggested benzimidazole resistance. Larval cultures showed that cyathostomes accounted for between 86 and 96% of pre-treatment parasite burdens at both sites. Other helminths identified in the faecal samples were Strongylus spp. and Trichostrongylus axei.     

Study on Synergistic Effect of VA5 Immunopotentiator on Pigeon NDV Inactivated Vaccine

This study was designed to evaluate the synergistic effect of VA5 immunopotentiator on pigeon ND4416 inactivated vaccine.160 healthy pigeons were randomly divided into four groups,including ND4416 strain inactivated vaccine group (NDV group),ND4416 strain inactivated vaccine and immunopotentiator (VA5) mixed group (NDV+VA5 group),La Sota inactivated vaccine group (La Sota group) and normal saline as the control group (C group),to assess their vaccine efficacy against virulent pigeon NDV by serological analysis and animal testing.Pigeons sera were collected at different time points after immunization and measured the HI antibody titer of each group.The results showed that VA5 immunopotentiator significantly improved the serum antibody level of pigeons immunized with pigeon Newcastle disease inactivated vaccine (P<0.05).In addition,comparative test of spleen lymphocyte transformation was conducted at various time points after immunization.The results indicated that VA5 effectively stimulated the lymphocyte transformation of immune pigeon.Pigeons in each groups were challenged with ND4416 strain at the 30th,90th and 180th d after immunization.The results presented that the NDV+VA5 group had 100% protection rate and higher than La Sota group.The duration of immunization test showed that the antibody titer of NDV+VA5 group reached peak 11.20log₂ at the 21st d,remained 7.50log₂ at 180th d,and the protection rate remained 100% at 180th d.It indicated that VA5 immunopotentiator sustained the immune duration of pigeon NDV vaccine up to 180 d.Moreover,the in vitro detoxification test results suggested that VA5 immunopotentiator reduced the in vitro detoxification cycle of pigeons after challenge.Overall,this study suggested VA5 immunopotentiator could significantly improve the immune efficacy of pigeon Newcastle disease inactivated vaccine,which provided a basis for further enhancing the efficacy of Newcastle disease vaccine,as well increased experimental data for the application of immunopotentiators.     

VA5免疫增强剂对鸽新城疫病毒灭活疫苗的增效作用研究

试验旨在探究VA5免疫增强剂对鸽新城疫病毒（NDV）灭活疫苗（ND4416株）的免疫增效作用。选取160只健康鸽随机分为4组：ND4416株灭活疫苗组（NDV组）、ND4416株灭活疫苗与免疫增强剂（VA5）混合组（NDV+VA5组）、La Sota灭活疫苗组（La Sota组）及生理盐水空白对照组（C组）,进行免疫效力及免疫持续期试验。采用血凝抑制试验检测各组鸽免疫后不同时间点的血清抗体效价,结果显示,VA5免疫增强剂能够显著提高鸽NDV灭活疫苗血清抗体水平（P<0.05）。脾脏淋巴细胞转化试验结果显示,VA5能够有效刺激免疫鸽的淋巴细胞转化。在免疫后第30、90、180天,各组鸽子采用ND4416株进行攻毒试验,结果显示,NDV+VA5组在3个时间点的攻毒保护率均为100%,均高于La Sota组。免疫持续期试验结果显示,NDV+VA5组血清抗体水平在免疫后第21天达峰值11.20log₂,第180天仍能达到7.50log₂,攻毒保护率达100%,表明VA5免疫增强剂可延长鸽新城疫疫苗的免疫效力持续期至180 d。攻毒后体外排毒检测结果表明,VA5免疫增强剂能够缩短攻毒后鸽的体外排毒周期,减少体外排毒。综上,VA5免疫增强剂能够显著提高鸽NDV灭活疫苗的免疫效果,为研制效果更好的鸽NDV疫苗提供了基础依据,同时也为免疫增强剂的应用增加了试验数据。     

Colonization of neonatal puppies by staphylococci.

The composition of the normal staphylococcal flora of bitches and their litters held in a breeding unit was studied. The animals were sampled at a number of sites using moistened swabs. Six bitches were sampled daily, for 10 days, before whelping and then, together with four puppies per litter, at whelping (day 11) and at 1 and 7 days thereafter. Staphylococcus intermedius formed the predominant staphylococcal isolate. S. intermedius counts at the oral and nasal sites on the bitches did not change markedly before whelping and remained low (< 6 cfu/swab). Significant rises in the oral counts on both the bitches (P < 0.05) and puppies (P < 0.001) were then observed after whelping (days 11-18). Abdominal counts on both the bitches and puppies also rose (P < 0.001) after whelping. S. intermedius counts at the vaginal vestibulum of the pregnant bitches were found to be higher than at any other site sampled and did not alter markedly until whelping when a decrease (P = 0.05; days 10-12) was observed. S. intermedius was not found at the anal site in any of the six bitches and only transiently colonized five of the puppies.     

Bacteria profiles in four coastal areas of East Zealand--coliform bacteria, E. coli and vibriolike organisms (VLO) (author's transl)]

The prevalence of E. coli, coliforms and Vibrio-like-organisms has been studied at four different sites along the east coast of Zealand. High values were registered in sediment taken from the water-line and at a depth of one meter. Generally there exist significant differences between the bacterial counts of water and sediment, and it is obvious that the greatest concentration is in the waterline sediment. The epidemiological and hygienic consequences of these facts are discussed.