An assessment of the duration of Mycoplasma hyopneumoniae infection in an experimentally infected population of pigs

Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary pathogen of enzootic pneumonia (EP), a highly prevalent respiratory disease that affects pigs worldwide. Previous studies have demonstrated that M. hyopneumoniae infection can be longer than 185 days; however, the total duration of infection has not been determined yet. Therefore, the objective of this study was to determine the duration of M. hyopneumoniae infection in asymptomatic carriers. To achieve our goal, 60 pigs were inoculated with M. hyopneumoniae strain 232 and the persistence of M. hyopneumoniae in the respiratory tract was assessed by detection of the bacterial DNA in bronchial swabs and the ability of the infected pigs to transmit the pathogen to sentinels. Infection of the inoculated animals was demonstrated by the detection of M. hyopneumoniae DNA in nasal swabs, seroconversion to the bacteria and the onset of dry coughing. Experimentally infected pigs shed M. hyopneumoniae prior to and after the cough was observed. M. hyopneumoniae DNA was detected in 100% of experimentally infected pigs at 94 days post infection (dpi), 61% at 214dpi and 0% at 254dpi. Experimentally infected pigs transmitted the bacteria to sentinels at 80 and 200dpi. Results of this study have demonstrated that M. hyopneumoniae infected pigs can be incubatory as well as convalescent carriers of the pathogen and that convalescent carriers can remain infectious for up to 200 days. Total clearance of M. hyopneumoniae in the group was evidenced, demonstrating that duration of M. hyopneumoniae infection lasts less than 254 days.     

Rapid methodology for antigenic profiling of FMDV field strains and for the control of identity, purity and viral integrity in commercial virus vaccines using monoclonal antibodies

Monoclonal antibodies (MAbs) developed against different foot-and-mouth disease virus (FMDV) vaccine strains were extensively used to study any possible antigenic variations during vaccine production in Argentine facilities. Additionally, a typing ELISA using strain specific MAbs was developed to detect potential cross contaminations among FMDV strains in master and working seeds with high specificity and sensitivity and to confirm strains identity in formulated vaccines. This assay was carried out for the South American strains currently in use in production facilities in Argentina (A24/Cruzeiro, A/Argentina/01, O1/Campos and C3/Indaial) and for the strain O/Taiwan, produced only for export to Asia. These non-cross reactive MAbs were also used to analyze the integrity of viral particles belonging to each one of the individual strains, following isolation of 140S virions by means of sucrose density gradients from the aqueous phase of commercial polyvalent vaccines. Antigenic profiles were defined for FMDV reference strains using panels of MAbs, and a coefficient of correlation of reactivity with these panels was calculated to establish consistent identity upon serial passages of master and production seeds. A comparison of vaccine and field strain antigenic profiles performed using coefficients of correlation allowed the rapid identification of two main groups of serotype A viruses collected during the last FMD epidemic in Argentina, whose reactivity matched closely to A/Argentina/2000 and A/Argentina/2001 strains.     

Comparison of agar gel immunodiffusion test, rapid slide agglutination test, microbiological culture and PCR for the diagnosis of canine brucellosis

The performance of the rapid slide agglutination test, with and without 2-mercaptoethanol (RSAT and 2ME-RSAT) and agar gel immunodiffusion test (AGID) was evaluated for the diagnosis of brucellosis in naturally infected dogs. The microbiological culture, PCR and clinical parameters were used as reference. A total of 167 dogs were clinically examined and tested by blood culture, culture of semen/vaginal swab and PCR in blood and semen/vaginal swab. According to the results observed the 167 dogs were divided into three groups: Brucella canis infected dogs (Group 1), B. canis non-infected dogs (Group 2) and dogs with suspected brucellosis (Group 3). The dogs were then tested by RSAT, 2ME-RSAT and AGID. Groups 1 and 2 were used to calculate the diagnostic sensitivity and specificity of the serological tests and the results observed in Group 3 were also discussed. The diagnostic sensitivity of RSAT, 2ME-RSAT and AGID was respectively 70.58%, 31.76%, and 52.94%. The diagnostic specificity of RSAT, 2ME-RSAT and AGID was respectively 83.34%, 100%, and 100%. In dogs with suspected brucellosis 15% were RSAT positive, none was 2ME-RSAT positive and 5% were AGID positive. Although the serological tests are the most commonly used methods for brucellosis diagnosis, a significant proportion of false-negative results were observed highlighting the importance of the direct methods of diagnosis, like blood culture and PCR to improve the diagnosis of canine brucellosis.     

Effect of the bovine viral diarrhoea virus (BVDV) infection on dairy calf rearing

The aim of this study was to compare the cumulative incidence of mortality, clinical diarrhoea and respiratory disease in calves, during their first six months of age, in herds with different bovine viral diarrhoea virus (BVDV) infection status. Calves’ health indicators were tested by comparing proportions in 101 farms with dissimilar infection condition. The results indicate that there was a significant relationship between the BVDV status (actively infected herd or not) and the cumulative incidence of mortality and respiratory disorders.     

Removal of Chromium, Copper, and Nickel from an Electroplating Effluent Using a Flocculent Brewer’s Yeast Strain of Saccharomyces cerevisiae

The release of heavy metals in aquatic systems due to the discharge of industrial wastewaters is a matter of environmental concern. Heat-inactivated cells of a flocculent strain of Saccharomyces cerevisiae were used in the bioremediation, in a batch mode, of a real electroplating effluent containing Cu, Ni, and Cr. In this approach, no previous reduction of Cr(VI) to Cr(III) was required. Cr(VI) was selectively removed (98%) by yeast biomass at pH 2.3. At this pH, Cr(VI) is mainly in the form of HCrO ₄ ^? and yeast surface is surrounded by H⁺ ions, which enhance the Cr(VI) interaction with biomass binding sites by electrostatic forces. Subsequently, pH of the effluent was raised up to 6.0; this pH maximizes the efficiency of cations removal since at this pH the main binding groups of yeast cells are totally or partially deprotonated. The passage of effluent through a series of sequential batches, at pH 6.0, allowed, after the third batch, the removal of Cu(II), Ni (II), Cr total, and Cr(VI) in the effluent to values below the legal limit of discharge. The strategy proposed in the present work can be used in plants for the treatment of heavy metals rich industrial effluents containing simultaneously Cr(VI) and Cr(III).     

The influence of European and American wild germplasm in hop (<Emphasis Type="Italic">Humulus lupulus</Emphasis> L.) cultivars

Microsatellite variation at the nuclear and chloroplast genomes was evaluated for wild European and wild American hops, in order to assess the genetic diversity and origin of cultivated hops. Seven nuclear loci and 32 chloroplast loci were used in the analysis of 182 hop accessions including wild European (68), wild American (48), and cultivars (66). A total of 116 alleles were identified using 7 nuclear microsatellites showing different averages of polymorphism and distribution in the wild American and European accessions and cultivars. Two main groups were established as revealed by several statistical analyses; one including European wild accessions and cultivars and a second group consisting of American wild accessions. Three polymorphic chloroplast microsatellite loci were detected, six alleles were scored which defined a total of five haplotypes that were exclusive or presented different distribution between American and European wild accessions. A major influence of the wild European haplotypes was detected among hop cultivars. To the best of our knowledge, this is the first work reporting the use of chloroplast microsatellites in hops.     

Essential oils for preventative treatment and control of Asian soybean rust

The objective of the work was to evaluate the effect of essential oils as a preventive treatment to control Asian soybean rust. Initially the fungitoxic effect of the essential oils of Hyptis marrubioides, Aloysia gratissima and Cordia verbenacea was tested on the urediniospores of Phakopsora pachyrhizi, through in vitro tests. The in vivo test was set up in a greenhouse, using cultivar MGBR-46. The treatments consisted of the three oils at different concentrations, a fungicide based on pyraclostrobin + epoxyconazole and a control. To verify the potential of the essential oils in preventive treatment, inoculation was conducted at 0, 6, 12 and 24?h intervals, after application of the treatments. All treatments inhibited 100% of Phakopsora pachyrhizi germination. In the in vivo test, it was observed that all of the oils were efficient in controlling soybean rust, by preventive treatment, mainly at the higher concentrations. Efficiency of the oils was reduced with the increase of the interval between the application of the treatments and the onset of the pathogen. The essential oils, at the tested dosages, were not as efficient as the pyraclostrobin + epoxyconazole based fungicide.     

Quantitative modification of the testicular structure in pigs fed with anabolic doses of clenbuterol

Morphological and structural data of the testes of thirty male pigs were recorded in order to evaluate the effects of an anabolic clenbuterol treatment. Pigs aged 6 months were randomly allocated to one of three experimental groups. In two treated groups, the animals were fed with anabolic doses of clenbuterol (1 ppm); in the CLB group (n = 10) clenbuterol was given until they were slaughtered (treatment period = 3 months) whereas in the CLBW group (n = 10) the clenbuterol was withdrawn two weeks before slaughter (treatment period = 2.5 months); clenbuterol was not given to the pigs of the control group (n = 10). Stereological estimations of the tissular volume fraction and tubular volume density were applied to quantify the structural constituents of the testes. The results showed an increased volume fraction of the testicular interstitium especially in the Leydig cell population, as a result of the clenbuterol treatment. The increase in the nuclear volume fraction of the Leydig cells was the more persistent change in the variations recorded in both treated groups with respect to the control. A regression of the seminal epithelium was also recorded in the treated animals. The rest of the structural parameters were closer to the normal figures in the CLBW group, suggesting a recovery of the testicular structure when clenbuterol was withdrawn.     

Thermal decomposition and isomerization of cis-permethrin and beta-cypermethrin in the solid phase

The stability to heart of cis-permethrin and beta-cypermethrin in the solid phase was studied and the decomposition products identified. Samples heated at 210 degrees C in an oven in the dark showed that, in the absence of potassium chlorate (the salt present in smoke-generating formulations of these pyrethroids), cis-permethrin was not isomerized, although in the presence of that salt, decomposition was greater and thermal isomerization occurred. Other salts of the type KXO3 or NaXO3, with X being halogen or nitrogen, also led to a considerable thermal isomerization. Heating the insecticides in solution in the presence of potassium chlorate did not produce isomerization in any of the solvents assayed. Salt-catalysed thermal cis-trans isomerization was also found for other pyrethroids derived from permethrinic or deltamethrinic acid but not for those derived from chrysanthemic acid. The main thermal degradation processes of cis-permethrin and beta-cypermethrin decomposition when potassium chlorate was present were cyclopropane isomerization, ester cleavage and subsequent oxidation of the resulting products. Permethrinic acid, 3-phenoxybenzyle chloride, alcohol, aldehyde and acid were identified in both cases, as well as 3-phenoxybenzyl cyanide from beta-cypermethrin. A similar decomposition pattern occurred after combustion of pyrethroid fumigant formulations.     

In-vitro evidence for activative thiolysis and selfsynergism of sulfenyl dicarbamate derivatives of 3,4 methylenedioxyphenyl N-methylcarbamate

Activative thiolytic cleavage of the N-S bond and self-synergism in sulfenyl dicarbamate (SD) derivatives of 3,4-methylenedioxyphenyl N-methylcarbamate has been investigated. The formation of precursor carbamates by the reaction of the propoxur derivative with glutathione and cysteine was demonstrated by HPLC. The slower rate of insectidal action of SD-propoxur compared with propoxur and its lower ki for inhibition of housefly acetylcholinesterase measured after preincubation of housefly head homogenate with N-ethylmaleimide were considered as indirect evidence of the N-S breakdown as a delay factor in toxicity. The SD-carbaryl derivative, 3,4-methylenedioxyphenyl N-methylcarbamate, and piperonyl butoxide similarly inhibited the oxidative metabolism of [¹⁴C]carbaryl in post-mitochondrial homogenates of housefly abdomens. The results indicated that activative thiolysis and self-synergism performed by the methylenedioxyphenyl group could be processes involved in the insecticidal action of the sulfenyl dicarbamates.