Determination of furaltadone and nifursol residues in poultry eggs by liquid chromatography-electrospray ionization tandem mass spectrometry

The use of nitrofurans as veterinary drugs has been banned from intensive animal production in the European Union (EU) since 1993. The objective of the present study was to evaluate the accumulation and depletion of furaltadone and nifursol and their side-chain metabolites 5-methylmorpholino-3-amino-2-oxazolidinone (AMOZ) and 3,5-dinitrosalicylic acid hydrazide (DNSAH) in eggs after administration of therapeutic and subtherapeutic doses of the drugs to laying hens during three consecutive weeks. LC-MS/MS, with positive and negative electrospray ionization methods, was used for the determination of parent compounds and metabolites in yolk and egg white and was validated according to criteria established by Commission Decision 2002/657/EC. The decision limit (CCα) and the detection capability (CCβ) of the analytical methodology for metabolites were 0.1 and 0.5 μg/kg for AMOZ and 0.3 and 0.9 μg/kg for DNSAH, respectively. For the parent compounds, CCα and CCβ were 0.9 and 2.0 μg/kg for furaltadone and 1.3 and 3.1 μg/kg for nifursol, respectively. The data obtained show that the parent compounds are much less persistent than their side-chain metabolites in either yolk or egg white. Between the studied metabolites, AMOZ is the most persistent and could be detected in either yolk or egg white three weeks following withdrawal from treatment.     

Use of crude, FML and rK39 antigens in ELISA to detect anti-Leishmania spp. antibodies in Felis catus

Visceral leishmaniasis is a disease caused by Leishmania (Leishmania) chagasi and represents a serious public health problem. The dog is the main urban reservoir of the disease; however, investigations regarding the occurrence and epidemiological importance of leishmaniasis in cats have recently been initiated. This study aimed to detect cats seropositive for Leishmania spp. using different antigens. Additional studies were performed using sera from cats with Toxoplasma gondii (n=15) to evaluate cross-reactivity. Serum samples (n=113) from cats living in the town of Ara?atuba, State of S?o Paulo, Brazil, an endemic area for human and canine visceral leishmaniasis, were tested by indirect ELISA using different antigens: crude (CAG-ELISA), fucose-mannose ligand (FML-ELISA) and K39 (rK39-ELISA). Anti-Leishmania spp. antibodies were detected in 23.0% of samples evaluated by CAG-ELISA, 13.3% by FML-ELISA and 15.9% by RK39-ELISA. Only reactive sera in all three tests were considered truly positive. No disagreement occurred among the tests (p<0.05). Serum samples seropositive for toxoplasmosis tested by CAG-ELISA were negative, but one sample (6.7%) was positive for FML-ELISA and rK39-ELISA suggesting a cross-reaction between these antigens and anti-T. gondii antibodies. These findings indicate the occurrence of feline leishmaniasis in Ara?atuba. Further studies are required to clarify the role of cats in the epidemiological cycle of leishmaniasis.     

Genotyping of potentially zoonotic Giardia duodenalis from exotic and wild animals kept in captivity in Brazil

We have studied the variability of glutamate dehydrogenase (gdh) and small subunit ribosomal (SSU) rRNA coding genes of Giardia species in fecal samples isolated from wild and exotic animals in Brazil, and compared with homologous sequences of isolates from human and domestic animals characterized in previous studies. Cysts of Giardia duodenalis were obtained from feces of naturally infected monkeys (Alouatta fusca) (n=20), chinchillas (Chinchilla lanigera) (n=3), ostriches (Struthio camelus) (n=2) and jaguar (Panthera onca) (n=1). Assemblage AI was assigned to the unique isolate of jaguar. All the samples from monkeys, chinchillas, and ostriches were assigned to Assemblage B. There was little evolutionary divergence between the referred isolates and isolates described elsewhere. The Assemblage B isolates identified in this study were closely related to Assemblage BIV isolated from humans. The molecular identification of Assemblages A and B of G. duodenalis isolates from exotic and wild animals demonstrates that such hosts may be a potential reservoir for zoonotic transmission of G. duodenalis.     

Brain anatomy of two-toed sloth (Choloepus didactylus,Linnaeus, 1758): A comparative gross anatomical study of extant xenarthrans

The neural system plays an important role in understanding some features of animals. Anatomical complexity correlates with the increase of functional capacity. Xenarthrans include anteaters (Vermilingua), armadillos (Cingulata) and sloths (Folivora). This group is the base of eutherian mammals, and understanding the anatomy of its neural system could provide data for functional and evolutionary interpretations. The gross anatomy of the xenarthran brain is recorded. Four extant families of Pilosa and two families of Cingulata were sampled. Usual dissection procedures were used, and the brains were analysed macroscopically. The brain of two-toed sloth, three-toed sloth, six-banded armadillo, giant anteater and collared anteater are gyrencephalic. Pygmy anteater, nine-banded armadillo, great long-nosed armadillo, southern naked-tailed armadillo and giant armadillo are lissencephalic. In most species, the rhinal fissure presents two segments, rostral and caudal (except in Vermilingua and three-toed sloth). The diencephalon and brainstem present similar anatomy. The cerebellum is wide and presents four lobes (rostral, central, caudal and floccular). Its average volume is 12.16% (Folivora), 14.26% (Vermilingua) and 18.61% (Cingulata). Among these groups, there is a statistical difference between Folivora/Cingulata concerning the cerebellum average. The general pattern of the brain of the xenarthrans is similar to that of other mammals.     

Digit innervation of the thoracic limb of Criollo horses: Anatomical description and consequences to perineural blocks

Criollo horse breeding is an important economic activity in South America. Because of their athletic performance, these animals tend to show great incidence of musculoskeletal disorders, many of them diagnosed by means of perineural blocks. However, incorrect interpretation of these blocks may be due to anatomical differences in nerve distribution. The objective of this study was to describe the innervation of the digit region of thoracic limbs in Criollo horses, in order to improve the interpretation of tests for claudication diagnosis based on nerve block. Thirty thoracic limbs from Criollo horses were dissected. It could be observed that in 90% of the limbs, dorsal branches of the palmar nerve originated proximally to the proximal sesamoid bone. In 93% of the cases, the palmar digital nerve and the dorsal branches communicated; in 87% of the cases, communication between branches of the dorsal branch was observed; and in 27% (8/30) of the limbs, the palmar metacarpal nerve and the dorsal branch presented communications. None of the specimens showed complete symmetry in the distribution of nerves in contralateral limbs. The high frequency of communication between the nerves may be a particularity of the Criollo breed that may interfere with the interpretation of perineural blocks. Based on the anatomical position, it may be inferred that divergent results in Criollo horses may occur when abaxial sesamoid nerve block is used. Palmar digital nerve block may be less influenced by these variations, provided it is performed as distal as possible from the ungular cartilage.     

Composting Wastes Contaminated with Naphthalene

Recent data show composting as an alternative for treatment of PAHs contaminated wastes. Nevertheless, the presence of highly volatile compounds like naphthalene might compromise the interest of this treatment and thus its careful evaluation is justified. A bench scale composting system was built to evaluate the importance of volatilization in the transformation of naphthalene by composting. Naphthalene concentration was measured in solid, aqueous and gas phases by HPLC and GC. Preliminary results have shown that the presence of microorganisms able to degrade naphthalene reduces naphthalene loss by volatilization.     

Toxicological and metabolic parameters of the teleost fish (Leporinus obtusidens) in response to commercial herbicides containing clomazone and propanil

Pesticides, such as herbicides can affect the metabolic and toxicological parameters on fish. For this reason, an experiment was carried out with the objective of to evaluate the effects of commercial formulations of clomazone and propanil herbicides on acetylcholinesterase (AChE), thiobarbituric acid-reactive substances (TBARS), catalase (CAT) and metabolic parameters in teleost fish (Leporinus obtusidens). Fish were exposed during 90 days to field measured concentration of the herbicides clomazone and propanil (376 and 1644 μg/L, respectively) on rice paddy water. Specific AChE activity in the brain and muscle decreased and TBARS levels decreased in brain, muscle and liver tissues. Liver catalase decreased after exposure to both herbicides. Metabolic parameters in the liver and white muscle showed different changes after exposure to both herbicides. In summary, the results showed that clomazone and propanil affects toxicological and metabolic parameters of piavas. These results suggest that environmentally relevant herbicides concentrations are toxic to Leporinus obtusidens.     

A nontoxic derivative of Bothrops jararaca venom suitable to generate antibodies against the native venom.

Bothrops jararaca venom was toxoided by stepwise iodination with cold iodine, and doses up to 30 LD50 were non-lethal by i.p. route (BICALHO et al., 1990). Groups of mice injected chronically with the native, or the iodinated venom, have been subjected to histological examination. In the native group, in the spleen, around the white pulp, an acellular, amorphous eosinophilic substance, metachromatic to Methyl Violet, PAS positive, and dichroic to Congo Red under polarized light, was present. Strong congestion in the liver, kidneys and lungs was found. The salivary glands were replenished with an amorphous substance in the serosal acini. The groups injected with the iodinated venom only show discrete alterations, more akin to the control group. The anavenin was immunogenic. Antibody generation in mice and rabbits was detected by ELISA. In mice, active protection against challenge with native venom was presented. The iodinated venom generated a rabbit antiserum with strong lines in gel immunoprecipitation against the lethal venom. A minimum neutralization titer of 2.3 mg ml-1 against the native venom was attained in the first cycle (28 days) of immunization. After 3 cycles (100 days), the protection rose to 5.1 mg ml-1.     

The expression of plasmid mediated afimbrial adhesin genes in an avian septicemic Escherichia coli strain

An Escherichia coli strain (SEPT13) isolated from the liver of a hen presenting clinical signs of septicaemia had a LD₅₀ of 4.0 × 10⁵ CFU/ml in one-day-old chickens, expressed Ia, Ib, E1, E3, K and B colicins and aerobactin. The strain was ampicillin and streptomycin resistant, and found to have fimA, csgA and tsh DNA related sequences; it could adhere to and invade HEp-2 and tracheal epithelial cells, expressed fimbriae (observed by electron microscopy), and had five plasmids of 2.7, 4.7, 43, 56, and 88 MDa. Transposon mutagenesis of strain SEPT13, with transposon TnphoA, resulted in a mutant strain named ST16 that had a LD₅₀ of 1.2 × 10¹² CFU/ml. All other biological characteristics of strain ST16 were the same as those detected for strain SEPT13 except for the migration of an 88 MDa plasmid to the 93 MDa position indicating the insertion of the transposon into the 88 MDa plasmid. The 93 MDa plasmid of strain ST16 was transferred, by electroporation assay, to non-pathogenic receptor strains (E. coli strains K12 MS101 and HB101), resulting in transformant strains A and B, respectively. These strains exhibited adhesion properties to in vitro cultivated HEp-2 cells but did not have the capacity for invasion. The adherence occurred despite the absence of fimbriae; this finding suggests that the 88 MDa plasmid has afimbrial adhesin genes.