Effects of different lipid sources on intake, digestibility and purine derivatives in hair lambs.

An experiment was conducted to evaluate the effects of different lipid sources on the nutrient intake, digestibility and purine derivative excretion of lambs. Thirty-five 60-day-old, male, non-castrated Santa Ines lambs with an initial average body weight (BW) of 13.00 ± 1.80 kg were used in a randomized complete block design with seven blocks and five treatments. The experimental treatments consisted of a control diet without supplemental lipids and four test diets with different lipid supplements, selected according to the degree of ruminal protection from hydrogenation: supplementation, being supplementation with whole cottonseed (WC), supplementation with cashew nut meal (CNM), supplementation with both cottonseed and cashew nut meal (WC-CNM) and supplementation with calcium salts of long-chain fatty acids (Ca-LCFA). The lambs were slaughtered after reaching 28 kg average BW for each treatment. The ether extract intake (EEI) was higher (p < 0.01) for the lipid supplemented compared to control diet lambs. Supplementation with WC decreased the digestibility of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF) and total carbohydrate (TC) (p < 0.01), whereas supplementation with CNM, WC-CNM and Ca-LCFA reduced non-fibrous carbohydrate (NFC) digestibility (p < 0.01). The ether extract (EE) digestibility coefficient was higher with CNM, followed by Ca-LCFA and WC, when compared to WC-CNM and control diets. Nitrogen balance (NB) was not influenced (p > 0.05) by the different lipid sources. A lower purine derivative (PD) excretion and thus lower microbial protein supply (MPS) was observed for animals supplemented with Ca-LCFA (p < 0.01) compared to the WC-CNM and control diets. In conclusion, WC, CNM and WC-CNM supplementation did not have negative effects on MPS, although negative effects have been observed on nutrient digestibility.

Crystal structure of Bn IV in complex with myristic acid: a Lys49 myotoxic phospholipase A2 from Bothrops neuwiedi venom.

The LYS49-PLA2s myotoxins have attracted attention as models for the induction of myonecrosis by a catalytically independent mechanism of action. Structural studies and biological activities have demonstrated that the myotoxic activity of LYS49-PLA2 is independent of the catalytic activity site. The myotoxic effect is conventionally thought to be to due to the C-terminal region 111-121, which plays an effective role in membrane damage. In the present study, Bn IV LYS49-PLA2 was isolated from Bothrops neuwiedi snake venom in complex with myristic acid (CH3(CH2)12COOH) and its overall structure was refined at 2.2 Å resolution. The Bn IV crystals belong to monoclinic space group P21 and contain a dimer in the asymmetric unit. The unit cell parameters are a = 38.8, b = 70.4, c = 44.0 Å. The biological assembly is a "conventional dimer" and the results confirm that dimer formation is not relevant to the myotoxic activity. Electron density map analysis of the Bn IV structure shows clearly the presence of myristic acid in catalytic site. The relevant structural features for myotoxic activity are located in the C-terminal region and the Bn IV C-terminal residues NKKYRY are a probable heparin binding domain. These findings indicate that the mechanism of interaction between Bn IV and muscle cell membranes is through some kind of cell signal transduction mediated by heparin complexes.

Age-related ultrasonography, cytology, and microbiologic exam of canine prostate / Ultrassonografia, citologia e exame microbiológico da próstata de cães em diferentes idades

The physiological parameters that could be reference for trustful diagnosis and prognosis of prostate disorders in dogs were obtained. Thirty six intact male dogs without clinical signs of neither prostatic nor reproductive disorders were allocated according the age in three groups. These animals were submitted to semen manual collection for microbiological exams; transabdominal ultrasonography to evaluate dimensions, ecogenicity, and texture of prostatic parenchyma and aspirative puncture with fine needle for cytological and microbiological analyses. Ultrasonography revealed that the predominant prostatic shape was round with regular surface. Dimensions varied according to age, being small in young animals and large in old ones. There was a positive correlation between prostatic dimensions and body weight. Microbiological exams detected microorganisms on seminal plasma from 11 dogs and prostate tissue aspirated from 10 animals, although they were healthy. Cytology did not reveal any inflammatory, proliferative, or neoplasic alteration in young and middle age dogs, but in three older dogs signs of hyperplasia/hypertrophy was found. It was observed positive correlation between age and cellular area but a negative correlation was observed between nucleus:cytoplasm ratio and craniocaudal dimension.

Obtiveram-se parâmetros fisiológicos que pudessem ser utilizados como referência para diagnóstico e prognóstico confiáveis de doença prostática em cães. Trinta e seis cães, sem sinais clínicos de doença prostática ou distúrbios reprodutivos, foram distribuídos em três grupos de acordo com a idade.Os animais foram submetidos à colheita manual de sêmen para exames microbiológicos, à ultrassonografia transabdominal, para avaliar as dimensões, a ecogenicidade e a ecotextura prostática, e à punção aspirativa com agulha fina, para análise citológica e microbiológica. A ultrassonografia revelou que a forma predominante da próstata foi globosa, com superfície de contorno regular. As dimensões variaram de acordo com a idade, sendo pequena em animais jovens e grande nos animais idosos. Houve correlação positiva entre as dimensões prostáticas e o peso corporal. Os exames microbiológicos detectaram microrganismos no plasma seminal de 11 cães e no tecido prostático aspirado de 10 animais, embora eles fossem saudáveis. A citologia não revelou nenhuma alteração inflamatória, proliferativa ou neoplásica nos cães jovens e de meia idade, mas, em três cães idosos foram encontrados sinais de hiperplasia/hipertrofia. Foi observada correlação positiva entre a idade e a área celular e correlação negativa entre a relação núcleo:citoplasma e a dimensão craniocaudal.

Purification of a PHA-like chitin-binding protein from Acacia farnesiana seeds: a time-dependent oligomerization protein.

A lectin-like protein from the seeds of Acacia farnesiana was isolated from the albumin fraction, characterized, and sequenced by tandem mass spectrometry. The albumin fraction was extracted with 0.5 M NaCl, and the lectin-like protein of A. farnesiana (AFAL) was purified by ion-exchange chromatography (Mono-Q) followed by chromatofocusing. AFAL agglutinated rabbit erythrocytes and did not agglutinate human ABO erythrocytes either native or treated with proteolytic enzymes. In sodium dodecyl sulfate gel electrophoresis under reducing and nonreducing conditions, AFAL separated into two bands with a subunit molecular mass of 35 and 50 kDa. The homogeneity of purified protein was confirmed by chromatofocusing with a pI = 4.0 +/- 0.5. Molecular exclusion chromatography confirmed time-dependent oligomerization in AFAL, in accordance with mass spectrometry analysis, which confers an alteration in AFAL affinity for chitin. The protein sequence was obtained by a liquid chromatography quadrupole time-of-flight experiment and showed that AFAL has 68% and 63% sequence similarity with lectins of Phaseolus vulgaris and Dolichos biflorus, respectively.

Ion-exchange chromatography used to isolate a spermadhesin-related protein from domestic goat (Capra hircus) seminal plasma

Mammalian seminal plasma contains among others, proteins called spermadhesins, which are the major proteins of boar and stallion seminal plasma. These proteins appear to be involved in capacitation and sperm-egg interaction. Previously, we reported the presence of a protein related to spermadhesins in goat seminal plasma. In the present study, we have further characterized this protein, and we propose ion-exchange chromatography to isolate this seminal protein. Semen was obtained from four adult Saanen bucks. Seminal plasma was pooled, dialyzed against distilled water and freeze-dried. Lyophilized proteins were loaded onto an ion-exchange chromatography column. Dialyzed-lyophilized proteins from the main peak of DEAE-Sephacel were applied to a C2/C18 column coupled to an RP-HPLC system, and the eluted proteins were lyophilized for electrophoresis. The N-terminal was sequenced and amino acid sequence similarity was determined using CLUSTAL W. Additionally, proteins from DEAE-Sephacel chromatography step were dialyzed and submitted to a heparin-Sepharose high-performance liquid chromatography. Goat seminal plasma after ion-exchange chromatography yielded 6.47 +/- 0.63 mg (mean +/- SEM) of the major retained fraction. The protein was designated BSFP (buck seminal fluid protein). BSFP exhibited N-terminal sequence homology to boar, stallion and bull spermadhesins. BSFP showed no heparin-binding capabilities. These results together with our previous data indicate that goat seminal plasma contains a protein that is structurally related to proteins of the spermadhesin family. Finally, this protein can be efficiently isolated by ion-exchange and reverse-phase chromatography.

Acquired vesicovaginal fistula secondary to ovariohysterectomy in a bitch: a case report

Uma cadela de sete anos de idade foi trazida ao hospital veterinário "Governador Laudo Natel" pois apresentava estrangúria há dois meses. Os sinais clínicos desenvolveram-se dois dias após ovário-histerectomia de eleição. Exames radiográficos e ultrasonográficos sugeriram piometra de coto ou granuloma cervical e fistula vesicovaginal. Duas laparotomias foram realizadas para desfazer as adesões, mas não houve melhora nos sinais clínicos observados. Iniciou-se tratamento médico e oito meses depois o animal, novamente avaliado, apresentava-se sadio porém ainda com sinais de estrangúria.

Further studies on the structural analysis of the cuticle of Litomosoides chagasfilhoi (Nematoda: Filarioidea).

In order to obtain further information on the structural organization of the cuticle of nematodes, this structure was isolated from adult forms of the filariid Litomosoides chagasfilhoi. The purity of the fraction was determined by light and transmission electron microscopy, deep-etching, high resolution scanning electron microscopy, atomic force microscopy, immunocytochemistry, gel electrophoresis (SDS-PAGE) and Western blot. The epicuticle presented a rugous surface with parallel rows and several globular particles that could be involved in the absorption of nutrients and secretion of products. Analysis by SDS-PAGE of purified cuticles revealed five major polypeptides corresponding to 151, 41, 28, 13 and 11 kDa. A polyclonal antibody against a synthetic 18 amino-acid peptide that corresponds to the sequence of domain E of the Haemonchus contortus3A3 collagen gene recognized several protein bands on the Western blot of purified cuticle, and labeled all cuticular layers, as shown by immunocytochemistry.

Isolation and partial characterisation of a protein from buck seminal plasma (Capra hircus), homologous to spermadhesins.

Spermadhesins are a family of secretory proteins expressed in the male genital tract of pig, horse and bull. Their function and structure have been widely studied, especially those isolated from boar. However, there are no data concerning spermadhesins isolated from buck. Buck seminal plasma was collected and subjected to ion exchange chromatography on DEAE-Sephacel column followed by chromatography in a C18 column coupled to a HPLC system. The purification of the protein was determined by SDS-PAGE and MALDI-TOF analysis exhibiting a molecular mass of 12.5 KDa and showed to be structurally homologous to spermadhesins from boar and stallion.