In Vitro Activities of Antifungal Agents against Clinical Isolates of Dermatophytes from Animals

Although the susceptibility of dermatophytes to antifungal drugs is well documented in humans, the effectiveness in animals has not been previously investigated. The in vitro susceptibility of 54 clinical isolates from animal dermatophytoses to ketoconazole (KTZ), itaconazole (ITZ) and terbinafine (TFN) was measured using microdilution assay (CLSI M38-A2 test) and by the E-test (KTZ and ITZ). All 3 drugs showed antifungal activity, while KTZ displayed the broadest minimum inhibition concentration (MIC) range (0.125-16 μg/ml) against M. canis and M. gypseum. The MIC of KTZ and ITZ was almost the same for human and animal isolates of T. mentagrophytes and T. rubrum. The MIC of TFN was almost the same for dermatophytes isolated from humans and animals.     

Phylogenetic Analysis of Anaplasma marginale Strains from Paraná State,Brazil, Using the msp1α and msp4 genes

Anaplasma marginale is an obligate intraerythrocytic rickettsial pathogen (order, Rickettsiales: family, Anaplasmataceae) that causes bovine anaplasmosis. This disease is widely distributed in tropical and sub‐tropical regions of the world and causes important economic losses to cattle production. Major surface protein (MSP)1a (msp1α gene) is one of the six MSPs identified on A. marginale from cattle, whose sequence and size vary according to the number of tandem 28‐ to 29‐amino acid repeats. This study characterized the msp1α and msp4 genes obtained from three distinct Brazilian herds from the State of Paraná. Three strains of the msp1α and one strain of the msp4 gene were sequenced. The strains evaluated revealed PCR products of different size, representing three, five and six internal repeats. Sequence analyses confirmed the number of tandem sequence copies and revealed a high degree of sequence identity with strains from other Brazilian States, as well as strains from the USA, Europe and Israel. The msp1α DNA and amino acid sequences from A. marginale and DNA sequences of msp4 strains did not reveal distinct phylogeographical segregation. However, the amino acid sequences of msp4 demonstrated definite phylogeographical relationship. These results suggest that the amino acid sequences of msp4 should be used for phylogenetic identification of A. marginale strains and may be an important tool for the epidemiology and control of anaplasmosis. Additionally, the close similarity of the Paraná strains of A. marginale with strains from USA, Europe and Asia may reflect the introduction of these genes during the development of the Brazilian bovine herd.     

Arthroderma benhamiae infection in a rabbit

The isolate from the rabbit with dermatophytosis which was transmitted to the owners was proved to be Arthroderma benhamiae (-) by mating experiments as well as by chitin synthase 1 (CHSI) gene analysis.     

Molecular heterogeneity in clinical isolates of Malassezia pachydermatis from dogs

Molecular investigation of 16 strains, conventionally identified to be Malassezia pachydermatis, isolated from dogs in Japan was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD band patterns of 13 clinical isolates were identical to that of standard strain of M. pachydermatis (CBS-1879). The other three clinical isolates were different from the standard strain of M. pachydermatis in RAPD patterns, and two of the three isolates were identical. About 620 bp genomic DNA fragments of the CHS2 gene were amplified from the same 16 clinical isolates of M. pachydermatis by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of the 16 clinical isolates revealed that the 13 strains were genetically very close to the standard strain of M. pachydermatis and the other two isolates were genetically close to the standard strain of M. furfur rather than M. pachydermatis. The remaining one isolate was phylogenetically distinct from all the seven Malassezia species reported so far.     

Immune response of BALB/c mouse immunized with recombinant MSPs proteins of Anaplasma marginale binding to immunostimulant complex (ISCOM)

Anaplasmosis, caused by Anaplasma marginale, results in significant economic losses of cattle in tropical and subtropical regions worldwide. Six major surface proteins (MSPs) were well characterized and designated as MSP1, MSP2, MSP3, MSP4, and MSP5. The objective of this study was to evaluate the humoral immune response of BALB/c mice against the recombinant MSPs, incorporated into immunostimulating complex (ISCOM). The recombinant proteins purified by Ni-NTA columns were incorporated into ISCOM and ISCOMATRIX by the lipid film hydration method. BALB/c mice immunized with ISCOM/rMSPs and ISCOMATRIX/rMSPs vaccines produced whole IgG, IgG1, and IgG2a, in contrast to the negative groups (PBS and ISCOMATRIX adjuvant). All groups that received antigen responded specifically against the rMSPs by Western blotting, showing the rMSP1a (60-105kDa), rMSP1b (100kDa), rMSP4 (47kDa), and rMSP5 (29kDa). Additional studies will have to be performed in cattle to evaluate the humoral and cellular mechanisms of this subunit vaccine and their possible use as protective vaccines against homologous and heterologous strains of A. marginale.     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

Slipping magnetic reconnection in coronal loops

Magnetic reconnection of solar coronal loops is the main process that causes solar flares and possibly coronal heating. In the standard model, magnetic field lines break and reconnect instantaneously at places where the field mapping is discontinuous. However, another mode may operate where the magnetic field mapping is continuous but shows steep gradients: The field lines may slip across each other. Soft x-ray observations of fast bidirectional motions of coronal loops, observed by the Hinode spacecraft, support the existence of this slipping magnetic reconnection regime in the Sun's corona. This basic process should be considered when interpreting reconnection, both on the Sun and in laboratory-based plasma experiments.     

Aspergillus niger pulmonary infection in a dog

A 6-month-old male golden retriever was presented with fever, bloody-watery diarrhea and mild cough. Parvovirus and Isospora canis infection was confirmed and successfully treated. Two weeks later, the dog had severe cough and mucopurulent nasal discharge. Aspergillus niger was cultured from endotracheal washings on blood agar at 37 degrees C. Treatment with itraconazole for about 10 weeks resolved the clinical signs.