Histopathological and Immunohistochemical Characteristics of Thyroid Carcinoma in the Dog.

Thyroid carcinomas are a common form of endocrine neoplasia in dogs. In the present study, we combined histopathology with immunohistochemistry (IHC) to search for the presence of oestrogen receptor alpha (ORα), Cox-2 and Ki67 in canine thyroid carcinomas. Forty-eight thyroid carcinomas were diagnosed throughout the study period. Thyroglobulin and calcitonin IHC distinguished between thyroid tumours with a follicular and medullary (C-cell) origin, respectively. IHC-based diagnosis showed that 42 (87.50%) of the cases were follicular cell carcinoma. In these cases, the follicular-compact pattern was the most frequent (n = 20/42; 47.62%) and six cases (12.5%) were medullary cell (C-cell) carcinomas. Both medullary (C-cell) and follicular carcinomas expressed Ki67 and Cox-2. No differences were observed between medullary and follicular carcinomas with respect to expression of Ki67 (P = 0.34) and Cox-2 (P = 0.9523) markers. A total of 4.17% (n = 2/48) of thyroid carcinomas showed positive nuclear labelling for ORα, suggesting that oestrogen does not directly participate in the pathogenesis of canine thyroid neoplasia.

Pre-Multidrug-Resistant Mycobacterium tuberculosis Infection Causing Fatal Enteric Disease in a Dog from a Family with History of Human Tuberculosis.

This report describes a fatal case of a pet dog with major enteric signs owned by a family that has experienced cases of pulmonary tuberculosis (TB) in the household. Clinical and epidemiological aspects, imaging data, microbiological, haematological and histopathological examinations were assessed to diagnosis of disease. gyrB-RFLP, spoligotyping and MIRU-VNTR allowed molecular detection of M. tuberculosis strain from S family. The resazurin microtiter assay indicated that all isolates were resistant to isoniazid, ethambutol, ciprofloxacin, ofloxacin, streptomycin and amikacin. The public health concerns related to canine tuberculosis and risk of the dissemination by pets of M. tuberculosis pre-multidrug-resistant (PMD) to isoniazid, ethambutol and other first-line drugs used in human therapy of TB are discussed. We believe this to be the first report of PMD M. tuberculosis infection in a dog presenting mainly enteric manifestation, confirmed as S lineage by molecular methods, owned by a family in which TB has spread in the household for generations.

Fish composition and species richness in eastern South American coastal lagoons: additional support for the freshwater ecoregions of the world.

The relationships between fish composition, connectivity and morphometry of 103 lagoons in nine freshwater ecoregions (FEOW) between 2·83° S and 37·64° S were evaluated in order to detect possible congruence between the gradient of species richness and similarities of assemblage composition. Most lagoons included in the study were <2 km(2) , with a maximum of 3975 km(2) in surface area. Combined surface area of all lagoons included in the study was 5411 km(2) . Number of species varied locally from one to 76. A multiple regression revealed that latitude, attributes of morphometry and connectivity, and sampling effort explained a large amount of variability in species richness. Lagoon area was a good predictor of species richness except in low latitude ecoregions, where lagoons are typically small-sized and not affected by marine immigrants, and where non-native fish species accounted for a significant portion of species richness. Relationships between species and area in small-sized lagoons (<2 km(2) ) is highly similar to the expected number in each ecoregion, with systems located between 18·27° S and 30·15° S attaining higher levels of species richness. Similarities in species composition within the primary, secondary and peripheral or marine divisions revealed strong continental biogeographic patterns only for species less tolerant or intolerant to salinity. Further support for the FEOW scheme in the eastern border of South America is therefore provided, and now includes ecotonal systems inhabited simultaneously by freshwater and marine species of fishes.

A new species of Centromochlus (Siluriformes, Auchenipteridae, Centromochlinae) from the middle Rio Tocantins basin, Brazil.

A new species of the catfish genus Centromochlus (Auchenipteridae, Centromochlinae) is described. The new species is diagnosed by having numerous dark rounded blotches over the body and fins, dorsal-fin spine with serrations anteriorly and smooth posteriorly, anal fin of mature males with three unbranched and seven branched rays, anterior nuchal plate absent and posterior nuchal plate not extended ventrally. The new species is described from a small stream in the Estação Ecológica Serra Geral de Tocantins, a natural reserve in the centre of the Brazilian Cerrado, close to the watershed between the Rio Tocantins and the Rio São Francisco basins. The new species is possibly the sister taxon to the recently described Centromochlus meridionalis from the upper Rio Tapajós. Those two species share with Centromochlus perugiae, from the upper Amazon and upper Paraguay, derived features associated with the modified anal fin in sexually mature males.

Calcium and magnesium concentrations in mature human milk: influence of calcium intake, age and socioeconomic level.

Concentrations of calcium and magnesium were measured in mature milk, collected between 30 and 90 days after childbirth, from a group of 90 mothers between 14 and 39 years of age, exclusively breastfeeding. The group was divided into three sub-groups: low socioeconomic-level adolescents (LSAd), low socioeconomic-level adults (LSA), and high socioeconomic-level adults (HSA). Each mother's nutritional status was determined using the body-mass index (BMI) and her eating habits, obtained on the basis of a 24-h dietary recall. Adolescent and adult mothers in the low socioeconomic-level group had lower average calcium intake (LSAd = 618.4 +/- 555.2 mg and LSA = 679.4 +/- 411.4 mg) than adult mothers in the higher socioeconomic-level group (853.6 +/- 415.5 mg). The average concentration of calcium in the adolescent mothers' milk (LSAd) was significantly lower (5.30 +/- 1.42 mmol Ca/L, P = 0.01) than that of the two adult groups (LSA = 5.82 +/- 1.55 mmol Ca/L and HSA = 6.40 mmol Ca/L). The average magnesium concentrations for all groups did not show significant differences (LSAd = 1.06 +/- 0.18, LSA = 1.16 +/- 0.23 and HSA = 1.11 +/- 0.23 mmol Mg/L, for P= 0.16). These results indicate that magnesium concentrations in mature human milk do not seem to depend on maternal nutritional status. The condition of adolescence, however, associated with lower calcium intake by the mother, resulted in lower calcium concentrations in the milk secreted when compared to that of adult mothers.

Survey of Brazilian tomato products for alternariol, alternariol monomethyl ether, tenuazonic acid and cyclopiazonic acid.

Alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TEA) are secondary metabolites of Alternaria species. Cyclopiazonic acid (CPA) is produced by fungi belonging to the ubiquitous genera of Penicillium and Aspergillus and has been found in a wide range of foods. These toxins were searched for by liquid chromatography with diode-array detection in tomato products processed and sold in Brazil. Eighty samples of tomato products were examined (juice 11, pulp 22, purée 22, paste 24 and whole stewed tomato 1). Eleven brands sold nationwide were covered. TEA was found in seven samples of tomato pulp (39-111 ng/g) and four samples of tomato puree (29-76 ng/g). CPA was found in six samples of pulp (64-178 ng/g) and two samples of purée (36-117 ng/g). Co-occurrence of TEA and CPA was found in two samples of purée and one of pulp. This is the first time that the presence of CPA has been reported in tomato products. Neither AME nor AOH were detected in the samples.

Cutaneous myoepithelioma: an under-recognized cutaneous neoplasm composed of myoepithelial cells.

Benign and malignant neoplasms of myoepithelial cells comprise a rare but well-characterized group of tumors, among which myoepithelioma of the salivary glands is the best known. Extrasalivary examples of myoepithelioma also have been described in the breast, larynx, and retroperitoneum. Recently, myoepithelioma of the soft tissue also has been reported. According to this description, myoepithelioma and mixed tumors arising in the skin and subcutis represent points along a clinicopathologic spectrum of cutaneous and soft-tissue tumors. To the best of our knowledge, there has been only one case report of an entirely cutaneous myoepithelioma in the literature. We report herein five additional examples of purely myoepithelial tumors located exclusively in the dermis. Histopathologically, the neoplasms were well-circumscribed dermal lesions composed of fascicles of spindle cells with eosinophilic cytoplasm and ovoid-to spindle-shaped nuclei. Focally, neoplastic aggregations of more epithelioid cells representing large round cells with abundant pale cytoplasm arranged in solid clusters, cords, or strands were also seen. Ductal differentiation was not identified in either of these solid aggregations of epithelioid cells or in the fascicles of spindle-shaped cells. Nuclear pleomorphism in epithelioid and spindle-cell areas was mild, and mitotic figures were very sparse. In some cases, small, necrotic areas were seen within the solid aggregations of spindle-shaped cells. Neoplastic stroma was scant and composed of fibrillary collagen and abundant mucin. In one case, the stroma consisted of clusters of mature adipocytes intermingled with fascicles of myoepithelial cells. Areas of chondroid or osteoid metaplasia were not seen in any of the cases. Immunohistochemically, neoplastic cells expressed positivity for muscle specific actin (HHF35), alpha smooth muscle actin (IA4), S-100 protein, glial fibrillary acidic protein (GFAP), and epithelial membrane antigen (EMA), whereas stains for pan-cytokeratin (MNF116) were focal and weak. The findings in this report expand the clinical and histopathologic spectrum of cutaneous myoepithelioma, an under-recognized cutaneous neoplasm of myoepithelial cells.

Ochratoxin A in Brazilian roasted and instant coffees.

Thirty-four samples of roast and ground coffee, 14 samples of instant coffee and two samples of decaffeinated instant coffee were collected in markets and supermarkets in the city of Campinas, Brazil, and analysed for ochratoxin A using immunoaffinity columns for clean-up and HPLC with fluorescence detection for quantification. The limit of detection was 0.2 ng/g ochratoxin A. Twenty-three samples of ground and roast coffee were found to be contaminated with the toxin at levels ranging between 0.3 and 6.5 ng/g. The average concentration in all 34 samples was 0.9 ng/g. All samples of instant coffee contained ochratoxin A at levels ranging from 0.5 to 5.1 ng/g, with an average figure of 2.2 ng/g. Roast and ground coffee is the type of coffee most used by Brazilians for the preparation of the beverage. Considering that an average Brazilian adult takes five cups of coffee per day, which corresponds to 30 g of roast and ground coffee, the probable daily intake of ochratoxin A by a 70 kg adult would be 0.4 ng/kg bw, which is far below the current Provisional Tolerable Daily Intake of 14 ng/kg bw for ochratoxin A as set by the Codex Alimentarius. To study the transfer of ochratoxin A into coffee brew, the beverage was prepared by two methods: (a) the drip method and (b) the Brazilian country style method. No significant difference was observed between the two methods in terms of extraction of the toxin using five contaminated samples containing between 0.8 and 6.5 ng/g ochratoxin A. The drip method extracted 86 +/- 15% and the Brazilian country style 74 +/- 20% of the ochratoxin A initially present in the roast and ground coffee.

Fumonisins B1 and B2 in Brazilian corn-based food products.

Eighty-one samples of corn products were acquired from markets and supermarkets in the city of Campinas, SP, Brazil, and were analysed for fumonsins B1 and B2 (FB1 and FB2). Forty samples (49%) were positive for FB1 (0.03-4.93 (micrograms/g) and 44 samples (54%) for FB2 (0.02-1.38 (micrograms/g). The samples, in order of decreasing contamination, were, corn meal (all contaminated, 0.56-4.93 (micrograms/g FB1), followed by degerminated corn (8/11 samples, nd-4.52 (micrograms/g FB1), corn flour (9/11 samples, nd-1.46 (micrograms/g FB1), precooked corn flour (4/6 samples, nd-1.79 (micrograms/g FB1), corn grits (2/2 samples, 0.17-1.23 (micrograms/g FB1), and popcorn (4/9 samples, nd-1.72 (micrograms/g FB1). Relatively lower incidences and levels of contamination were found in corn flakes (1/4 samples, nd-0.66 (microgram/g FB1) and corn flour baby cereal (1/2 samples, nd and 0.44 (microgram/g FB1). The samples of corn on the cob (common corn in the milky stage, 7 samples) and of the typical foods 'curau' (2 samples) and 'pamonha' (7 samples), both prepared with corn in the milky stage, did not show any detectable contamination. Canned sweet corn, also harvested in the milky stage, exhibited a very low incidence of and level of contamination (2/11 samples, nd-0.08 (microgram/g FB1). The intake of corn products is low in urban areas and in most rural areas in Brazil. In certain rural areas, however, corn products play a greater role in daily meals and the calculated intake of FB1 is higher than a proposed Tolerable Daily Intake of 800 ng/kg bw/day. This is the first report on fumonisins in Brazilian corn-based food products.

Fumonisin levels in commercial corn products in Buenos Aires, Argentina.

Fumonisins B1 (FB1) and B2 (FB2) were determined in 35 samples of corn flour and corn grits destined for human consumption and purchased directly from Buenos Aires food shops and supermarkets from October 1996 to January 1997 and during the month of January 1998. During the first period of sample collecting, 16 out of 19 samples were found to be contaminated. Considering all 19 samples, contamination levels were between not detected and 1860 ng/g FB1, and from not detected to 768 ng/g FB2. During the second period all 16 samples were found to be contaminated with levels ranging from 75 to 4987 ng/g FB1, and from not detected to 1818 ng/g FB2. The levels of FB1 and FB2 in the samples collected during January 1998 were significantly higher than the samples collected during the period from October 1996 to January 1997. No significant difference was found in terms of fumonisin levels between the branded and unbranded samples.

Poisoning by Diacetoxyscirpenol in cattle fec citrus pulp in the State of Sao Paulo, Brazil

Relata-se a morte de bovinos causada pela ingestäo de polpa cítrica contaminada pela micotoxina Diacetoxiscirpenol (7mg/kg), pertencente ao grupo dos tricotecenos

Mycotoxins and fungi in wheat stored in elevators in the state of Rio Grande do Sul, Brazil.

Samples of wheat harvested from 1988 to 1990 and stored in elevators in the south of Brazil (12 Brazilian, 4 Argentinian and 2 Uruguayan) were analysed in 1990 for 14 mycotoxins: deoxynivalenol (DON), nivalenol, diacetoxyscirpenol (DAS), T-2 and HT-2 toxins, T-2 triol, T-2 tetraol, aflatoxins B1, B2, G1, G2, ochratoxin A (OCHRA A), zearalenone and sterigmatocystin. One sample (1988 harvest) was contaminated with OCHRA A (0.04 microgram/g) and three other samples (1990 harvest) were contaminated with DON (0.40 microgram/g), DAS (0.30 microgram/g), T-2 (two samples, 0.35 and 0.36 gamma g/g) and T-2 tetraol (1.68 micrograms/g). Fusarium graminearum Schwabe was found in the 1990 samples with a relative incidence ranging from 1 to 22% and predominated in Argentinian and Uruguayan wheat (1990 harvest). Fusarium dimerum Penzig (8-75%) was the main Fusarium sp. in Brazilian wheat from the 1990 harvest.

Mycotoxins and fungi in wheat harvested during 1990 in test plots in the state of São Paulo, Brazil.

Wheat from two cultivars with contrasting characteristics were harvested in ten experimental plots located in wheat producing areas of the State of São Paulo, Brazil. The samples (10 of each cultivar) were analyzed by a gas-chromatographic method for deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), toxins T-2 (T-2) and HT-2, T-2 tetraol, T-2 triol, and by a thin-layer chromatographic method for zearalenone (ZEN), aflatoxins B1, B2, G1, G2, ochratoxin A and sterigmatocystin. No mycotoxins were detected in 13 samples. DON was found in four samples (0.47-0.59 microgram/g), NIV in three samples (0.16-0.40 microgram/g), T-2 in two samples (0.40, 0.80 microgram/g), DAS in one sample (0.60 microgram/g), and ZEN in three samples (0.04-0.21 microgram/g). The wheat samples were also examined for the incidence of fungi. Alternaria, Drechslera, Epicoccum and Cladosporium were the prevailing genera. Among the Fusarium spp., F. semitectum was present in 19 samples and F. moniliforme in 18 samples. No F. graminearum was isolated in the samples.

Survey of aflatoxins, ochratoxin A, zearalenone, and sterigmatocystin in some Brazilian foods by using multi-toxin thin-layer chromatographic method.

A previously published method for ochratoxin A was evaluated and proved appropriate for simultaneous determination of aflatoxins, ochratoxin A, sterigmatocystin, and zearalenone, with considerable savings in time and reagent costs. The detection limits were 2, 5, 15, and 55 micrograms/kg, respectively. The recoveries and coefficients of variation obtained with artificially contaminated samples were 91-101% and 0-16% for aflatoxin B1, 98-117% and 0-17% for sterigmatocystin, and 96-107% and 0-17% for zearalenone, respectively. The coefficients of variation for naturally contaminated samples (aflatoxins in rice and ochratoxin A in beans) ranged from 0 to 8%. The method was used to survey 296 samples that included 10 cultivars of dried beans, 8 types of corn products, 3 types of cassava flour, and both polished and parboiled rice between May 1985 and June 1986 in Campinas, Brazil. Only aflatoxin B1 (9 samples, 20-52 micrograms/kg), aflatoxin G1 (4 samples, 18-31 micrograms/kg), and ochratoxin A (5 samples, 32-160 micrograms/kg) were found. The average contamination percentage was 4.7%; beans showed the highest (6.6%) and rice showed the lowest (3.3%) incidence rates. Zearalenone and sterigmatocystin were not detected. Positive samples were confirmed by chemical derivatization, corroborated by development in 3 solvent systems.

Screening and quantitation of ochratoxin A in corn, peanuts, beans, rice, and cassava.

To answer the need for simple, economical, rapid methods for mycotoxins, a procedure for screening and quantitation of ochratoxin A was developed. A methanol-aqueous KCl extraction is used, followed by cleanup with clarifying agents and partition into chloroform. Part of the chloroform extract is used for screening and the other part for quantitation by thin layer chromatography (TLC). The screening procedure takes 40 min, using a silica gel/aluminum oxide minicolumn developed for this purpose. The limits of detection are 80 and 10 micrograms/kg, respectively, for minicolumn screening and TLC quantitation. Ammonium sulfate is efficient in cleaning samples of corn and cassava; cupric sulfate is better with peanuts, beans, and rice. Tests were conducted on triplicate spiked samples of yellow corn meal, raw peanuts, dried black beans, polished rice, and cassava flour at different levels (400, 200, 80, 40, and 10 micrograms/kg). Recoveries ranged from 86 to 160% and the coefficients of variation ranged from 0 to 26%.