Experimental infection of dogs (Canis familiaris) with sporulated oocysts of Neospora caninum

Neospora caninum is widely distributed in the world and this parasite is one of the major causes of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum and several species of domestic and wild animals are intermediate hosts. Dogs can become infected by the ingestion of tissues containing cysts and then excrete oocysts. It is not yet known whether sporulated oocysts are able to induce a patent infection in dogs, i.e. a shedding of N. caninum oocysts in feces. The objective of this study was to experimentally examine the infection of dogs by sporulated oocysts. The oocysts used in the experiment were obtained by feeding dogs with brain of buffaloes (Bubalus bubalis) positive for anti-N. caninum antibodies by indirect fluorescent antibody test (IFAT ≥200). Oocysts shed by these dogs were confirmed to be N. caninum by molecular methods and by bioassay in gerbils, and sporulated N. caninum oocysts were used for the oral infection of four dogs. The dogs were 8 weeks old and negative for antibodies to N. caninum and Toxoplasma gondii. Dogs 1 and 4 received an inoculum of 10,000 sporulated oocysts each; dog 2 an inoculum of 5000 sporulated oocysts and dog 3 received 1000 sporulated oocysts of N. caninum. The total feces excreted by these dogs were collected and examined daily for a period of 30 days. No oocysts were found in their feces. The dogs were monitored monthly for a 6-month period to observe a possible seroconversion and when this occurred the animals were eliminated from the experiment. Dogs 1 and 4 seroconverted 1 month after the infection with titer, in the IFAT, of 1600 and 800, respectively; the other two dogs presented no seroconvertion during the 6-month period. Dogs 1 and 2 were euthanized 180 days after infection and were examined for the detection of N. caninum in tissues (brain, muscle, lymph node, liver, lung, heart and bone marrow) by immunohistochemistry and PCR with negative results in both techniques. Bioassay in gerbils with brain of these dogs was also performed and again the results were negative. In conclusion, dogs infected with sporulated oocysts of N. caninum were not able to shed oocysts in feces. However, a higher dose of infection stimulated the production of antibodies against N. caninum in the dogs.     

Determination of tetracycline antibiotic residues in edible swine tissues by liquid chromatography with spectrofluorometric detection and confirmation by mass spectrometry

A sensitive and specific method is described for the simultaneous determination of oxytetracycline, tetracycline (TC), and chlortetracycline residues in edible swine tissues, by combining liquid chromatography with spectrofluorometric and mass spectrometry detection. The procedure involved a preliminary extraction with EDTA-McIlvaine buffer acidified at pH 4.0, followed by solid-phase extraction cleanup using a polymeric sorbent. The liquid chromatography analysis was performed with spectrofluorometric detection after postcolumn derivatization with magnesium ions. The limits of quantification were 50 microg/kg for muscle and 100 microg/kg for kidney tissues. The recovery values were greater than 77.8% for muscle and 65.1% for kidney. The method has been successfully used for the quantification of tetracyclines in swine tissues samples. The selective liquid chromatography mass spectrometric analysis for confirmation of oxytetracycline in one positive swine muscle sample was made by atmospheric pressure chemical ionization (APCI). The APCI mass spectra of the TCs gave the protonated molecular ion and two typical fragment ions, required for their confirmation in single ion monitoring scan mode in animal tissues.     

Viability of Extended Goat Semen Stored at Refrigerated Condition

Due to the small volume of goat semen ejaculate, just a few doses of goat semen were produced when the sperm concentration is 100 × 10^/mL. The study was aimed to determine the viability of extended goat semen at refrigerated condition at 5 ℃ using varying sperm concentrations and evaluated if sperm concentration lower than 100 × 10^6/mL would affect the motility, viability and sperm morphology at refrigerated condition. Using an artificial vagina, ejaculated goat semen was collected from goat semen donor aged 1.5 year. Physical evaluation of the collected semen showed an average volume of 0.54 mL, mean pH of 6.8 and a milky white color with thick consistency indicative of high concentration. Fresh goat semen had an initial average of 76% with an average initial sperm concentration of 128 × 10^/mL. The semen was divided into four treatments： sperm concentration of 100× 10^/mL, 75 × 10^/mL, 50 × 10^/mL and 25 × 10^/mL, and were stored at 5 ℃ for a period of 10 d. The semen evaluation was performed for each of the four treatments every other day. Results showed that the sperm concentration of spermatozoa affected the duration of storage based on the sperm motility percentage, viability and morphology of spermatozoa. The extended goat semen with sperm concentration of 25 and 50 million sperm/mL is optimal for storage within 6 d that gave satisfactory percentage motile, viable and morphologically normal spermatozoa.     

Persistent Mullerian Duct Syndrome in a Miniature Schnauzer Dog with Signs of Feminization and a Sertoli Cell Tumour

A 5‐year‐old male Miniature Schnauzer was presented with unilateral cryptorchidism and signs of feminization. Abdominal ultrasonography revealed an enlarged right testis and a large, fluid‐filled cavity that appeared to arise from the prostate. Computed tomography revealed the cavity to be consistent with an enlarged uterine body, arising from the prostate, and showed two structures resembling uterine horns that terminated close to the adjacent testes. The dog had a normal male karyotype, 78 XY. Gonadohysterectomy was performed and both the surgical and the histological findings confirmed the presence of a uterus in this male animal, resulting in a diagnosis of persistent Mullerian duct syndrome (PMDS). The enlarged intra‐abdominal testis contained a Sertoli cell tumour. Computed tomography proved to be an excellent diagnostic tool for PMDS.     

Tree girdling provides insight on the role of labile carbon in nitrogen partitioning between soil microorganisms and adult European beech

Nitrogen (N) cycling in terrestrial ecosystems is complex since it involves the closely interwoven processes of both N uptake by plants and microbial turnover of a variety of N metabolites. Major interactions between plants and microorganisms involve competition for the same N species, provision of plant nutrients by microorganisms and labile carbon (C) supply to microorganisms by plants via root exudation. Despite these close links between microbial N metabolism and plant N uptake, only a few studies have tried to overcome isolated views of plant N acquisition or microbial N fluxes. In this study we studied competitive patterns of N fluxes in a mountainous beech forest ecosystem between both plants and microorganisms by reducing rhizodeposition by tree girdling. Besides labile C and N pools in soil, we investigated total microbial biomass in soil, microbial N turnover (N mineralization, nitrification, denitrification, microbial immobilization) as well as microbial community structure using denitrifiers and mycorrhizal fungi as model organisms for important functional groups. Furthermore, plant uptake of organic and inorganic N and N metabolite profiles in roots were determined.Surprisingly plants preferred organic N over inorganic N and nitrate (NO₃⁻) over ammonium (NH₄⁺) in all treatments. Microbial N turnover and microbial biomass were in general negatively correlated to plant N acquisition and plant N pools, thus indicating strong competition for N between plants and free living microorganisms. The abundance of the dominant mycorrhizal fungi Cenococcum geophilum was negatively correlated to total soil microbial biomass but positively correlated to glutamine uptake by beech and amino acid concentration in fine roots indicating a significant role of this mycorrhizal fungus in the acquisition of organic N by beech. Tree girdling in general resulted in a decrease of dissolved organic carbon and total microbial biomass in soil while the abundance of C. geophilum remained unaffected, and N uptake by plants was increased. Overall, the girdling-induced decline of rhizodeposition altered the competitive balance of N partitioning in favour of beech and its most abundant mycorrhizal symbiont and at the expense of heterotrophic N turnover by free living microorganisms in soil. Similar to tree girdling, drought periods followed by intensive drying/rewetting events seemed to have favoured N acquisition by plants at the expense of free living microorganisms.     

Genetic diversity of Toxoplasma gondii isolates from chickens from Brazil

Until recently, Toxoplasma gondii was considered clonal with very little genetic variability. Recent studies indicate that T. gondii isolates from Brazil are genetically and biologically different from T. gondii isolates from USA and Europe. In the present study, we retyped 151 free range chicken isolates from Brazil including 117 newly isolated samples from 11 geographically areas (Alagoas, Bahia, Ceará, Maranh?o, Paraná, Pernambuco, Rio de Janeiro, Rio Grande do Norte, S?o Paulo, Sergipe, and Rondonia) and 34 previously reported isolates from the very north (Pará) and the very south (Rio Grande do Sul). Ten PCR-RFLP markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico were used to genotype all isolates. Overall analysis of 151 T. gondii isolates revealed 58 genotypes. Half (29/58) of these genotypes had single isolate and the other half of the genotypes were characterized with two or more isolates. Only 1 of 151 isolates was clonal Type I strain and 5 were clonal Type III strains. Two isolates had mixed infections. Clonal Type II strain was absent. One strain was Type II at all loci, except BTUB. The results confirm high genetic diversity of T. gondii isolates from Brazil.     

Subnanometer-Diameter Wires Isolated in a Polymer Matrix by Fast Polymerization

The preparation and analysis of inorganic-organic polymer nanocomposites consisting of inorganic nanowires and multiwire "cables" in a random-coil organic polymer host is reported. Dissolution of inorganic (LiMo3Se3)n wires in a strongly coordinating monomer, vinylene carbonate, and the use of a rapid polymerization in the presence of a cross-linking agent produce nanocomposites without phase separation. Polymerization of dilute solutions yields a material containing mostly (Mo3Se3(-))n mono- and biwires, 6 to 20 angstroms in diameter and 50 to 100 nanometers long. Polymerization of more concentrated liquid crystalline solutions yields a nanocomposite containing oriented multiwire cables, 20 to 40 angstroms in diameter and up to 1500 nanometers long, that display optical anisotropy and electrical conductivity.