Efficacy of 2,6-dichlorophenol lure to control Dermacentor nitens

This study was carried out with the objective of evaluating the efficacy of a 2,6-dichlorophenol (2,6-DCP) lure to control Dermacentor nitens (Acari: Ixodidae). Slow-release formulations of the pheromone formulated with and without cypermethrin were prepared. Olfactometer bioassays were used to define the best dose of the pheromone and to evaluate the effect of cypermethrin with 2,6-DCP attractiveness. Sexually active males were released 15 cm from 2 cmx1 cm pieces of polypropylene treated with different odors: 2,6-DCP in a liposphere system (1.5, 30 and 300 microg--without cypermethrin and 30 microg--with cypermethrin) and 2,6-DCP in hexane (30 microg). The tests were conducted 7 and 15 days after the preparation of the odor sources. The percentages of males that oriented, or showed directional movement toward the stimulus, and their tracks were recorded. Lures (10 cmx2 cm pieces of polypropylene) treated with the best dose of the liposphere formulation (30 microg) were prepared. The lures were evaluated in horses that had been artificially infested with D. nitens. The horses' ears were infested with 3000 D. nitens larvae per ear, once weekly for 4 weeks. The animals were divided into three groups: control, 2,6-DCP and 2,6-DCP+cypermethrin. On day 0, the lures of their respective treatments were attached to the horses' napes. From days 6 to 20 after attachment, female ticks of 4 mm or over in length were counted on the ears, every 2 or 3 days. Olfactometer analysis showed higher orientation to 30 microg dose and more prolonged release of the pheromone in the liposphere formulation than in hexane; cypermethrin did not interfere with the attractive effect of the pheromone. The lures were efficient in the first 10 days after attachment, when the mean number of females was higher in the control group (24.9) than in 2,6-DCP and cypermethrin (5.4), and 2,6-DCP (9.2) groups. After that period the number of females was similar in the control and treated groups. These results indicate that 2,6-DCP lures used in a liposphere formulation can control D. nitens for at least 10 days by preventing its copulation. However, further evaluation of longer-term pheromone release under natural conditions is needed in order to validate this kind of control. In addition, the use of extra lures on the horse's tail may help to control populations on the hindquarters.     

Conidiobolomycosis in sheep in Brazil

Conidiobolomycosis is reported in the state of Piauí, in the semiarid region of northeastern Brazil. Affected sheep had depression, weight loss, serous or mucohemorrhagic nasal discharge, and cranium-facial asymmetry from exophthalmos of 1 eye, generally with increased volume of the eyeball, keratitis, and corneal ulceration. At necropsy of 60 sheep, friable masses were observed in the posterior region of the nasal cavity, often destroying the ethmoturbinate bones. Frequently, the lesions invaded the nasal sinuses, cribiform plate, orbit, and brain. The masses were irregular, granular with moist surfaces, and soft and friable with white, yellow, or tan coloration. Dissemination of the lesion to lungs was observed in 27 sheep, to the brain in 26, to lymph nodes in 3, to the kidney in 2, and to the gallbladder and heart in 1. The microscopic examination showed granulomatous inflammation composed of central necrosis surrounded by lymphocytes, epithelioid and giant cells, and fibrous tissue. In all lesions, negatively stained structures representing hyphae were surrounded by Splendore-Hoeppli material. Coagulative necrosis, thrombosis, and vasculitis were also observed. Grocott methenamine silver stain showed 8-30-microm-thick hyphae, rarely septate or ramified, irregular in shape, and with black contoured wall, sometimes with bulbous dilatation in the extremities. On electron microscopy, the hyphae had a thick double wall surrounded by cellular remnants and an inflammatory exudate. Conidiobolus coronatus was isolated from the lesions of 6 sheep. Conidiobolomycosis is an important disease of sheep in the state of Piauí, and other regions of northeastern Brazil.     

Epidemiological aspects of canine babesiosis in the semiarid area of the state of Minas Gerais, Brazil

Epidemiological aspects of Babesia vogeli infection were studied in the canine population of a rural town located in the Brazilian “Drought Polygon” of the state of Minas Gerais, Brazil. The survey was carried out in March 2003, when 505 dogs were identified and their characteristics registered on appropriate forms. Blood samples were collected at this time and again in June, September and December 2003. Serum samples were tested by the indirect fluorescent antibody test (IFAT) to detect antibodies against B. vogeli. The prevalence of anti-B. vogeli antibodies was 18.8%; however, no correlations were found between prevalence of infection and the age or gender of the animals. Cross-bred dogs presented a higher chance of acquiring infection in comparison to pure-bred dogs. Significant differences concerning the incidence of the disease were found during the period April–June in comparison to other months, demonstrating that transmission of B. vogeli is related to seasonal variations of tick infestations. The results indicate that climatic factors within the semiarid area interfere directly in the epidemiology of canine babesiosis.     

Antimicrobial resistance in Enterococcus spp. and Escherichia coli isolated from poultry feed and feed ingredients

Poultry feed is at the start of the food safety chain in the "farm-to-fork" model, and might serve as a source of antimicrobial resistant bacteria present in poultry meat. Antimicrobial resistance was investigated in 1137 enterococci and 163 Escherichia coli strains recovered from 23 samples of commercial broiler feed and 66 samples of raw feeding materials taken over half a year timespan. Enumeration of enterococci and E. coli were also performed using traditional plating and fluorescent in situ hybridisation methods. Viable enterococci were detected in all feed samples and in 66% of samples of separate feed ingredients, while E. coli was present in 50% and 32% of feed and raw feeding materials, respectively. The median values (50th percentile) for plate and FISH counts for feeds were, respectively, 2.70 log CFU/g and 5.52 log cells/g for enterococci, and 0.15 log CFU/g and 6.00 log cells/g for E. coli. Among enterococci recovered from feed ingredients, resistance to rifampicin, erythromycin, nitrofurantoin, tetracycline, and ciprofloxacin was found in 59.8%, 21.6%, 21.2%, 18.0% and 6.9% of the isolates, respectively. A considerable proportion of the enterococci isolates obtained from broiler feed displayed resistance to tetracycline (69.1%), rifampicin (58.5%), erythromycin (52.9%) and nitrofurantoin (36.2%). Lower percentage of resistance was observed to chloramphenicol (4.6%), ciprofloxacin (3.9%), vancomycin (1.9%) and ampicillin (1.2%). Among E. coli recovered from feed ingredients and poultry feeds, resistance to ampicillin, tetracycline and streptomycin was found in 22.9%, 27.6% and 19.0% and in 22.4%, 41.4% and 17.0% of the isolates, respectively. These data show that feedstuffs and poultry feeds are extensively contaminated by resistant enterococci and, in a lesser extent, by E. coli, thus leading to their introduction in the farm environment.     

Genetic variation of foot-and-mouth disease virus isolates recovered from persistently infected water buffalo (Bubalus bubalis)

Genetic variation of foot-and-mouth disease virus (FMDV) isolates, serotype O, recovered serially over a 1-year period from persistently infected buffalos was assessed. The persistent state was established experimentally with plaque-purified FMDV, strain O(1)Campos, in five buffalos (Bubalus bubalis). Viral isolates collected from esophageal-pharyngeal (EP) fluids for up to 71 weeks after infection were analyzed at different times by nucleotide sequencing and T(1) RNase oligonucleotide fingerprinting to assess variability in the VP1-coding region and in the complete genome, respectively. Genetic variation increased, although irregularly, with time after infection. The highest values observed for the VP1-coding region and for the whole genome were 2.5% and 1.8%, respectively. High rates of fixation of mutations were observed using both methodologies, reaching values of 0.65 substitutions per nucleotide per year (s/nt/y) and 0.44s/nt/y for nucleotide sequencing and oligonucleotide fingerprinting, respectively, when selected samples recovered at close time periods were analyzed. The data herein indicate that complex mixtures of genotypes may arise during FMDV type O persistent infection in water buffalos, which can act as viral reservoirs and also represent a potential source of viral variants. These results fit within the quasi-species dynamics described for FMDV, in which viral populations are constituted by related, non-identical genomes that evolve independently from each other, and may predominate at a given time.     

Proteomic survey of the pathogenic Mycoplasma hyopneumoniae strain 7448 and identification of novel post-translationally modified and antigenic proteins

Mycoplasma hyopneumoniae is an important pathogen for pigs, being the causative agent of enzootic pneumonia. Recently, the genome sequences of three strains, J, 7448 and 232 have been reported. Here, we describe the results of a proteomic analysis, based on two-dimensional gel electrophoresis of soluble protein extracts, immunoblot and mass spectrometry, which was carried out aiming the identification of gene products and antigenic proteins from the M. hyopneumoniae pathogenic strain 7448. A preliminary M. hyopneumoniae proteome map in two pH ranges (3-10 and 4-7) was produced. A total of 31 different coding DNA sequences (CDSs), including three hypothetical ones, were experimentally verified with the identification of the corresponding protein products by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. According to the Clusters of Orthologous Groups (COG) functional classification, the identified proteins were assigned to the groups of metabolism (13), cellular processes (5) and information and storage processing (4). Nine of the identified proteins were not classifiable by COG, including some related to cytoadherence and possibly involved in pathogenicity. Moreover, at least five highly antigenic proteins of M. hyopneumoniae were identified by immunoblots, including four novel ones (a heat shock protein 70, an elongation factor Tu, a pyruvate dehydrogenase E1-beta subunit and the P76 membrane protein). The now available proteome map is expected to serve as a reference for comparative analyses between M. hyopneumoniae pathogenic and non-pathogenic strains, and for methabolic studies based on cells cultured under modified conditions.     

Diagnosis of Canine Brucellosis: Comparison between Serological and Microbiological Tests and a PCR Based on Primers to 16S-23S rDNA Interspacer

A pair of primers directed to 16S-23S rDNA interspacer (ITS) was designed directed to Brucella genetic sequences in order to develop a polymerase chain reaction (PCR) putatively capable of amplifying DNA from any Brucella species. Nucleic acid extracts from whole-blood from naive dogs were spiked with decreasing amounts of Brucella canis RM6/66 DNA and the resulting solutions were tested by PCR. In addition, the ability of PCR to amplify Brucella spp. genetic sequences from naturally infected dogs was evaluated using 210 whole-blood samples of dogs from 19 kennels. The whole-blood samples collected were subjected to blood culture and PCR. Serodiagnosis was performed using the rapid slide agglutination test with and without 2-mercaptoethanol. The DNA from whole blood was extracted using proteinase-K, sodium dodecyl sulphate and cetyl trimethyl ammonium bromide followed by phenol–chloroform purification. The PCR was capable of detecting as little as 3.8 fg of Brucella DNA mixed with 450 ng of host DNA. Theoretically, 3.8 fg of Brucella DNA represents the total genomic mass of fewer than two bacterial cells. The PCR diagnostic sensitivity and specificity were 100%. From the results observed in the present study, we conclude that PCR could be used as confirmatory test for diagnosis of B. canis infection.     

Suppression of invasive lake trout in an isolated backcountry lake in Glacier National Park

Fisheries managers have implemented suppression programmes to control non‐native lake trout, Salvelinus namaycush (Walbaum), in several lakes throughout the western United States. This study determined the feasibility of experimentally suppressing lake trout using gillnets in an isolated backcountry lake in Glacier National Park, Montana, USA, for the conservation of threatened bull trout, Salvelinus confluentus (Suckley). The demographics of the lake trout population during suppression (2009–2013) were described, and those data were used to assess the effects of suppression scenarios on population growth rate (λ) using an age‐structured population model. Model simulations indicated that the population was growing exponentially (λ = 1.23, 95% CI: 1.16–1.28) prior to suppression. However, suppression resulted in declining λ (0.61–0.79) for lake trout, which was concomitant with stable bull trout adult abundances. Continued suppression at or above observed exploitation levels is needed to ensure continued population declines.     

The influence of the metacarpophalangeal joint angle on the transversal area and mean echogenicity of the superficial digital flexor tendon and suspensory ligament in gaited horses

The objective of this study was to assess the influence of the metacarpophalangeal (MCP) joint angle on the transversal area (TA) and mean echogenicity (ME) of the superficial digital flexor tendon (SDFT) and suspensory ligament (SL) in gaited horses. Ultrasound images were obtained from 50 healthy adult horses of the Mangalarga Marchador (MM; n=25) and Campeiro (n=25) breeds. Static and dynamic angles of the MCP joint were measured from photographs and video recordings. Higher ME values were evinced for the SL only in the group with the smaller dynamic angles of the MCP joint in the MM horses, in addition to weak negative correlation between the dynamic angle and ME. Moreover, weak negative correlation was also observed between the static angle and TA of the lateral branch of the SL and between the static angle and the ME of the SDFT. However, the difference observed in the group of MM horses, as well as the weak correlation, was not considered sufficient to support the hypothesis that a smaller angle of the MCP joint (greater extension) is associated with larger TA and ME values for the structures. The results also suggest that the static and dynamic angles of the MCP joint do not influence the TA and ME values of the SDFT and SL in gaited horses.