The control of Varroa destructor using oxalic acid

Twenty-four honeybee (Apis mellifera) colonies were used to monitor the efficacy of a solution of 2.9% oxalic acid (OA) and 31.9% sugar against the mite Varroa destructor. Mite mortality was established prior to and after OA treatments, which were conducted in August and September. The treatments resulted in 37% mite mortality as opposed to 1.11% in the controls. OA treatment conducted in September on previously untreated colonies resulted in 25% mite mortality. OA treatments in October and November resulted in approximately 97% mite mortality. These results suggest that OA is effective during the broodless period and less effective when applied to colonies with capped broods. The possible use of OA against the Varroa mite in honeybee colonies as an alternative to routine chemical treatments is discussed.     

Rotenone and oxalic acid as alternative acaricidal treatments for Varroa destructor in honeybee colonies

This experiment assessed the efficacy of rotenone and oxalic acid (OA) in an aqueous sugar solution in controlling the honeybee mite Varroa destructor. Colonies were populated with mite-infested brood combs and worker bees. Three rotenone or OA treatments administered during the period with capped brood on 31 July, 14 and 18 August resulted in an average efficacy of 24.10%. In untreated colonies mite mortality averaged 5.40%. No significant differences (P>0.05) were found between the rotenone and OA treatments. Three OA treatments administered on 9, 12 and 18 September resulted in a 77.93% mite mortality. An increase in mite drop (P<0.05) was observed at 2 and 4 days after each treatment. OA applications in broodless colonies resulted in significantly (P<0.001) higher mite mortality rates (98.65% average) than the three treatments of rotenone or OA in colonies with capped brood. The dynamics of mite mortality after each rotenone or OA treatment are discussed in this study.     

Efficacy of repeated trickle applications of oxalic acid in syrup for varroosis control in Apis mellifera: influence of meteorological conditions and presence of brood

Oxalic acid field trails for the control of varroosis (Varroa destructor) were carried out in an apiary located on the Mt. Imittos (Attica, Greece). The colonies received four successive applications (approximately one every 16 days) with 4.2% oxalic acid (OA) and 60% sugar solution by trickling method with two alternative types of syringes (an automatic self-filling dosing and a single-use) from the broodright to broodless period. The results indicate that the first three applications (from 6th October to 25th November-broodright period) resulted in 65.3% cumulative mite mortality, while only the last application (after the 26th November-broodless period) resulted in 77.3% mite mortality. Very low outern temperatures reduce to the minimum the bee movability, which may result into a slower development of the OA efficacy. No poor colony growth or queen loss were observed even if the bee colonies were received the four successive OA applications with the last one taken place at a very low outern temperature (6.2 degrees C). The trickling method using an automatic-filling syringe seems to be a very quick way for applying oxalic acid in large apiaries (approximately 150hives/h).     

Characterization of the immune response of domestic fowl following immunization with proteins extracted from Dermanyssus gallinae

Dermanyssus gallinae is the most significant ectoparasite of European poultry egg laying production systems due to high costs of control and associated production losses as well as adverse effects on bird welfare. In this study, soluble proteins were extracted from unfed D. gallinae (DGE) using a urea-based detergent and ultra-filtration, passed through a 0.22 μm filter and blended aseptically with adjuvant. One group of laying hens was immunized with DGE and adjuvant (Montanide ISA 50 V) whilst another group (Control) received physiological saline and adjuvant. All birds were immunized on two occasions, 21 days apart. Antibody response to immunization was determined by ELISA and western blotting using immunoglobulins (Igs) extracted from egg yolk. DGE immunization of hens resulted in a significant (P < 0.05) IgY response compared to controls, although there was no significant difference in IgM response between treatments. A number of proteins were identified by western blotting using IgY antibodies from DGE immunized birds, most prominently at 40 and 230 kDa. Analysis of proteins from approximately corresponding bands on SDS-PAGE confirmed the identity of tropomyosin, whilst other proteins showed high sequence homology with myosin and actin from other arachnid and insect species. Immunization of hens with DGE resulted in a 50.6% increase in mite mortality (P < 0.001) 17 h after feeding when tested by an in vitro mite feeding model. Data in this study demonstrate that somatic antigens from D. gallinae can be used to stimulate a protective immune response in laying hens. Further work is needed to identify other proteins of interest that could confer higher protection against D. gallinae, as well as optimization of the vaccination and in vitro testing protocol.     

Simulation modelling as a tool for evaluating surveillance programmes for detection of the Asian honeybee mite (Varroa destructor) in the South Island of New Zealand

AIM: To use a simulation model of the spread of the Asian honeybee mite (Varroa destructor) amongst apiaries, to evaluate a series of detection surveillance programmes for the South Island of New Zealand.

METHODS: Five potential incursion sites into the South Island were selected. A stochastic spatial simulation model, Varroa_sim, was adapted to simulate spread of the mite from these sites as a series of silent-phase propagating epidemics. The study population comprised all apiaries in the South Island registered in the Ministry of Agriculture and Forestry's (MAF's) apiary database in 2003. Six different surveillance programmes were simulated to try and detect the mite. Three of these were the actual multi-stage sampling plans conducted during the autumn (March–May) of 2001, 2002 and 2003, and the other three involved simple random sampling with sampling fractions equivalent to the actual numbers of apiaries tested in each of those years. The relative performances of the different surveillance plans were evaluated in terms of their ability to detect the mite early before it had spread too far and whilst there might still be a chance of eradication.

RESULTS: There were 13,798 registered apiaries in the South Island with valid map coordinates in the apiary database at the time of the study. The model generated 50 epidemics against which the various surveillance programmes were evaluated. The actual surveillance programmes conducted during the autumn of 2001 and 2002 generally performed fairly well in detecting the mite. The programme conducted in autumn 2003 detected the mite reasonably well in high-risk areas, but was very poor in low-risk areas. The simple random sampling strategies performed surprisingly well, and their relative rankings were proportional to the sampling fractions employed.

CONCLUSIONS: This study showed the value in using a spatial simulation model to generate plausible silent-phase epidemics, against which detection surveillance programmes could be evaluated, in ways that would otherwise not be possible.     

A new product with formic acid for Varroa jacobsoni Oud. control in Argentina. I. Efficacy

An organic product based on formic acid in a gel matrix was evaluated for use in Varroa control under autumnal climatic conditions in Argentina. Twenty colonies each received two gel packets with formic acid in two applications and numbers of falling mites were registered. After this treatment colonies received two other acaricides in order to compare efficacy. Average final efficacy in colonies treated with the organic product was 92% with a low variability. The gel matrix kept an adequate formic acid concentration inside the colonies with only two applications. This product is, therefore, a good alternative for Varroa control because it is organic, easy to use and presents a low variability in final efficacy between colonies. No queen, brood, or adult honeybee mortality was registered.     

Acaricidal effects of the essential oil of Origanum minutiflorum (Lamiaceae) against Rhipicephalus turanicus (Acari: Ixodidae)

The acaricidal effects of the volatile essential oil Origanum minutiflorum O. Schwarz & P.H. Davis (Lamiaceae) against adult Rhipicephalus turanicus was evaluated at a variety of concentrations and exposure times. Generally tick mortality increased with concentration and exposure. Ticks exposed to vapors from cotton wicks containing at least 10 μl/L resulted in complete (100%) mortality at 120 min. The major constituent of essential oil obtained from the plant material of O. minutiflorum was carvacrol.     

Occurrence of acute paralysis virus of the honey bee (Apis mellifera) in a Hungarian apiary infested with the parasitic mite Varroa jacobsoni.

Viruses of the honey bee have been known for a long time; however, recently the attention of scientists and apiculturalists has turned towards the relationship between these viruses and the parasitic mite Varroa jacobsoni. Although clinical symptoms indicated the presence of some of the viruses of bees in Hungary, none have previously been isolated or identified. During July unusual adult bee and brood mortality was observed in some colonies of an apiary in Budapest known to be infested with Varroa jacobsoni. Large amounts of acute paralysis virus (APV) were detected serologically in healthy honey bee pupae killed by the injection of a bacteria-free extract of diseased adult bees. Crystalline arrays of 30 nm particles were seen in ultrathin sections of the tissues of injected pupae and naturally infected adult bees. In spite of the application of acaricide treatments the bee population in several colonies had collapsed by the end of summer and the apiary suffered severe wintering losses.     

The efficacy of pyrantel pamoate against ascarids and hookworms in cats

The purposes of this study were to evaluate pyrantel pamoate administered orally at 20 mg/kg body weight for the removal of induced or natural infections of Ancylostoma tubaeformae and Toxocara cati in cats and to compare the efficacy of paste (40 mg base/g) and granule (80 mg base/g) formulations. Thirty cats of mixed breeding and various ages with natural and/or induced infections of A. tubaeformae and T. cati were assigned to one of three treatment groups: (1) non-medicated controls; (2) paste formulation at 20 mg base/kg; or (3) granule formulation at 20 mg base/kg. Infections were induced by feeding the cats on carcasses of infected mice. The study was conducted in replicates of at least one animal per treatment per replicate. The study parameters included clinical observations, physical examinations, faecal egg counts and the numbers, species and stages of worms recovered at necropsy. The paste formulation was 99.3% and 99.7% effective in reducing egg counts of Ancylostoma sp. and Toxocara sp. respectively. The granule formulation was 97.7% and 99.9% effective in reducing faecal egg counts of Ancylostoma sp. and Toxocara sp. respectively. When administered in paste form, pyrantel pamoate was 99.5% effective in removing adult Ancylostoma and 100.0% effective against adult Toxocara. The granule formulation was 97.9% effective against Ancylostoma and 100% effective against Toxocara. No toxic effects of either formulation of the drug were noted.     

Lack of prolonged activity of lavender essential oils as acaricides against the poultry red mite (Dermanyssusgallinae) under laboratory conditions

Managing the poultry red mite, Dermanyssus gallinae (De Geer) by conventional means (i.e., synthetic acaricides) has become increasingly problematic. As a possible alternative, research has identified several plant essential oils that are toxic to D. gallinae. However, essential oils are highly volatile and any acaricidal effect they exert could be short-lived in practice.This study investigated the short-lived toxicity of six lavender essential oils to D. gallinae. In sealed Petri-dishes, mites were exposed to filter papers impregnated with essential oil at a concentration of 0.14 mg/cm³. When filter papers were used immediately after impregnation, 66–90% D. gallinae mortality was observed after 24 h, depending upon the essential oil used. If impregnated filter papers were left in a fume cupboard for 24 h prior to use, mortality rates of D.gallinae fell to 11% or less.     

Isolation of <Emphasis Type="Italic">Vagococcus salmoninarum</Emphasis> in Rainbow Trout, <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> (Walbaum), Broodstocks: Characterization of the Pathogen

Coldwater ‘streptococcosis’, caused by Vagococcus salmoninarum, is an emerging disease of rainbow trout in the European Union, causing mortality rates up to 50% in broodstock during the spawning period, with water temperature of 10–12°C. A study to determine the presence and role of this bacterium was undertaken using classical bacteriological techniques confirmed with polymerase chain reaction. This is the first report of isolation of V salmoninarum in relation to outbreaks of mortality in a rainbow trout farm devoted exclusively to broodstock rearing in Spain. A total of 10 isolates of V salmoninarum were characterized by their morphological, cultural, physiological, biochemical and enzymatic traits. Some differences were observed in parameters such as growth on MacConkey agar, H2S production, acid production from starch, and some other minor variations. Isolates were sensitive to erythromycin and oxytetracycline tested in vitro, but treatments conducted in the field were ineffective. An attempt at vaccination did not provide encouraging results.     

Stimulation of the <Emphasis Type="Italic">In Vitro</Emphasis> Migration of Ovine Eosinophils by Factors Derived from the Sheep Scab Mite, <Emphasis Type="Italic">Psoroptes ovis</Emphasis>

The ectoparasitic astigmatid mite Psoroptes ovis causes sheep scab, a highly contagious, severe allergic dermatitis associated with damage to the fleece and hide, loss of condition and occasional mortality. The scab lesion is characterized by a massive infiltration of eosinophils that begins very rapidly after infection. This paper reports the finding that mite-derived factors directly enhance the migration of ovine eosinophils in vitro. Significant (p < 0.01) and dose-dependent (r = 0.972 ± 0.018 (SD)) activity was initially identified in whole mite extracts, by comparison with medium controls in an assay based on modified Boyden chambers and ovine bone marrow target cells. Similar pro-migratory activity (p < 0.005; r = 0.928 ± 0.069 (SD)) was detected in washes containing mite excretory/secretory material. By direct comparison with migration ratios (n = 3) for defined chemotactic (rmeotaxin = 3.430 ± 0.360 (SD)) and chemokinetic (rminterleukin-5 = 0.982 ± 0.112 (SD)) stimuli it was determined that the activity in both mite extracts (0.992 ± 0.038 (SD)) and mite washes (0.969 ± 0.071 (SD)) was chemokinetic. Subsequent experiments (n = 3) in which live mites were incorporated directly into the in vitro assay system indicated that they produced factors that significantly (p < 0.001) enhanced eosinophil migration to a degree directly related to mite numbers (r = 0.993 ± 0.005 (SD)). The identity of the factor(s) responsible is uncertain, but their presence suggests that mites may be capable of directly activating eosinophils in vivo, and raises the possibility that mites could directly influence, perhaps even initiate, the rapid early tissue eosinophilic response observed in experimental sheep scab infections.     

Surveillance of Toxigenic Fungi and Ochratoxin A in Feedstuffs from Córdoba Province,Argentina

The aim of this work was to evaluate the incidence of potential ochratoxigenic mycoflora and ochratoxin A (OA) in poultry, pig and rabbit feeds. Eighty poultry, pig and rabbit feed samples were taken at random from factories located from Córdoba province, Argentina, over a period of 8 months. Isolation and quantitative enumeration of fungal propagules were done on DRBC and DG18 media. The predominant species wereA. candidus, A. flavus,A. terreus,A. parasiticus,P. implicatum P. minioluteum,P. crustosum andP. citrionigrum. The distribution of sectionNigri species varied according to the feedstuffs analysed. The frequency ofA. niger var.niger was noticeably high in poultry feed samples on DRBC medium. TheNigri section species was present at moderate mean colony counts (CFU/g) from three feeds. Mycotoxin analysis of these samples showed that OA was detected in 15%, 10% and 12% of pig, poultry and rabbit feed samples, respectively. The mean levels detected ranged between 15 and 25 ng/g from three feeds. The presence of ochratoxigenic species ofNigri section and OA in feeds indicates the risk of potential exposure of poultry, pigs and rabbits through the ingestion of feeds.     

Establishment of canine and feline cells expressing canine signaling lymphocyte activation molecule for canine distemper virus study

Antimicrobial therapy is a useful tool to control bovine mastitis caused by Staphylococcus aureus, as consequence an increase in staphylococci resistant cases has been registered. Alternative strategies are desirable and bacteriocins represent attractive control agents to prevent bovine mastitis. The aim of this work was to evaluate the activity of five bacteriocins synthesized by Bacillus thuringiensis against S. aureus isolates associated to bovine mastitis. Fifty S. aureus isolates were recovered from milk composite samples of 26 Holstein lactating cows from one herd during September 2007 to February 2008 in México and susceptibility of those isolates to 12 antibiotics and 5 bacteriocins from B. thuringiensis was evaluated. S. aureus isolates were mainly resistant to penicillin (92%), dicloxacillin (86%), ampicillin (74%) and erythromycin (74%); whereas susceptibility to gentamicin, trimethoprim and tetracycline was detected at, respectively, 92%, 88%, and 72%. All S. aureus isolates showed susceptibility to the five bacteriocins synthesized by B. thuringiensis, mainly to morricin 269 and kurstacin 287 followed by kenyacin 404, entomocin 420 and tolworthcin 524. Our results showed that S. aureus isolates had differences in the antimicrobial resistance patterns and were susceptible to bacteriocins produced by B. thuringiensis, which could be useful as an alternative method to control bovine mastitis.     

Spatial epidemiology of the Asian honey bee mite (Varroa destructor) in the North Island of New Zealand

We describe the spatial epidemiology of Varroa destructor infestation among honey bee apiaries in the greater Auckland area of the North Island of New Zealand. The study population was comprised of 641 apiaries located within the boundaries of the study area on 11 April 2000. Cases were those members of the study population declared Varroa-infested on the basis of testing conducted between April and June 2000. The odds of Varroa was highest in apiaries in the area surrounding transport and storage facilities in the vicinity of Auckland International Airport. A mixed-effects geostatistical model, accounting for spatial extra-binomial variation in Varroa prevalence, showed a 17% reduction in the odds of an apiary being Varroa infested for each kilometre increase in the squared distance from the likely site of incursion (95% Bayesian credible interval 7–28%). The pattern of spatially autocorrelated risk that remained after controlling for the effect of distance from the likely incursion site identified areas thought to be ‘secondary’ foci of Varroa infestation initiated by beekeeper-assisted movement of infested bees. Targeted investigations within these identified areas indicated that the maximum rate of local spread of Varroa was in the order of 12 km/year (interquartile range 10–15 km/year).     

Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing

In total, 26 Campylobacter (C.) strains, isolated from liver, spleen, caecal or jejunal content of laying hens from different flocks were examined. In these flocks a drop in egg production, an increasing mortality and livers with whitish-grey lesions as post-mortem finding were observed. Suspected Campylobacter colonies were differentiated using a modified m-PCR in 13 Campylobacter jejuni and 13 Campylobacter coli strains. All isolates were characterised by typing of the flaA and flaB gene each with two restriction enzymes. To compare the four different profiles for all strains an artificial “fla-type” was generated. Different and identical fla-types of C. jejuni and C. coli were recovered from both intestinal and extra-intestinal organs of the laying hens and even from individual birds. One significant observation is that some fla-types of C. jejuni or C. coli were detected in intestinal and systemic sites but not all fla-types of both species appeared to be equally able to invade internal organs.     

Isolation,characterization and protection studies in mice of a streptomycin-dependent mutant ofStreptococcus suis type 1/2

An avirulent, streptomycin-dependent (Str-D) mutant ofStreptococcus suis type 1/2 was produced and characterized by its antimicrobial susceptibility, growth kinetics, biochemical reactions and reversion rate. Homologous and heterologous vaccine trials in mice resulted in complete protection against challenge withS. suis types 1 and 1/2 and partial protection against challenge withS. suis type 2.Abbreviations Str-D streptomycin-dependent - CFU colony-forming units - PBS phosphate-buffered saline - IP intraperitoneal - LD₅₀ the dose that results in 50% mortality     

Simulation modelling as a tool for evaluating surveillance programmes for detection of the Asian honeybee mite ( Varroa destructor ) in the South Island of New Zealand

AIM: To use a simulation model of the spread of the Asian honeybee mite (Varroa destructor) amongst apiaries, to evaluate a series of detection surveillance programmes for the South Island of New Zealand. METHODS: Five potential incursion sites into the South Island were selected. A stochastic spatial simulation model, Varroa_ sim, was adapted to simulate spread of the mite from these sites as a series of silent-phase propagating epidemics. The study population comprised all apiaries in the South Island registered in the Ministry of Agriculture and Forestry's (MAF's) apiary database in 2003. Six different surveillance programmes were simulated to try and detect the mite. Three of these were the actual multi-stage sampling plans conducted during the autumn (March-May) of 2001, 2002 and 2003, and the other three involved simple random sampling with sampling fractions equivalent to the actual numbers of apiaries tested in each of those years. The relative performances of the different surveillance plans were evaluated in terms of their ability to detect the mite early before it had spread too far and whilst there might still be a chance of eradication. RESULTS: There were 13,798 registered apiaries in the South Island with valid map coordinates in the apiary database at the time of the study. The model generated 50 epidemics against which the various surveillance programmes were evaluated. The actual surveillance programmes conducted during the autumn of 2001 and 2002 generally performed fairly well in detecting the mite. The programme conducted in autumn 2003 detected the mite reasonably well in high-risk areas, but was very poor in low-risk areas. The simple random sampling strategies performed surprisingly well, and their relative rankings were proportional to the sampling fractions employed. CONCLUSIONS: This study showed the value in using a spatial simulation model to generate plausible silent-phase epidemics, against which detection surveillance programmes could be evaluated, in ways that would otherwise not be possible.     

In vitro antiviral activity of chestnut and quebracho woods extracts against avian reovirus and metapneumovirus

Field evidences have suggested that a natural extract, containing tannins, could be effective against poultry enteric viral infections. Moreover previous studies have shown that vegetable tannins can have antiviral activity against human viruses. Based on this knowledge three different Chestnut (Castanea spp.) wood extracts and one Quebracho (Schinopsis spp.) wood extract, all containing tannins and currently used in the animal feed industry, were tested for in vitro antiviral activity against avian reovirus (ARV) and avian metapneumovirus (AMPV). The MTT assay was used to evaluate the 50% cytotoxic compounds concentration (CC₅₀) on Vero cells. The antiviral properties were tested before and after the adsorption of the viruses to Vero cells. Antiviral activities were expressed as IC₅₀ (concentration required to inhibit 50% of viral cytopathic effect). CC₅₀s of tested compounds were >200 μg/ml. All compounds had an extracellular antiviral effect against both ARV and AMPV with IC₅₀ values ranging from 25 to 66 μg/ml. Quebracho extract had also evident intracellular anti-ARV activity (IC₅₀ 24 μg/ml). These preliminary results suggest that the examined vegetable extracts might be good candidates in the control of some avian virus infections. Nevertheless further in vivo experiments are required to confirm these findings.     

香精油在蜂群抗螨中的应用

开发新的抗螨药物势在必行,而香精油及其组分具有显著的抑螨效果。百里酚(香精油的一种主要组分)及其一些组分,在实验室试验中其杀螨率一般都超过90%,而且对蜜蜂危害小,长期使用后在蜂蜜中的残留量很低,但要应用于养蜂生产,还有许多工作要做。对香精油在蜂群抗螨中的应用情况进行了综述。