Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase     

Nils H. Flores Huarco  Susana M. Giuliano  Fernanda G. Fumuso  Marcelo H. Miragaya  Hernán C. Cucho Dolmos  María I. Carretero 《Reproduction in domestic animals》2021,56(11):1387-1397

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.  相似文献   

Physiological and sanitary condition of the white clam Dosinia ponderosa collected from a coastal area impacted by shrimp farm effluent     

Marcel Martínez-Porchas  Susana Maria Scheuren-Acevedo  Luis Rafael Martínez-Córdova  Teresa Gollas-Galvan  Ramon H. Barraza-Guardado  Fernando Enríquez-Ocaña  Edilmar Cortés-Jacinto  Marco A. Porchas-Cornejo 《Aquaculture International》2016,24(1):243-256

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Effects of gamma radiation on the immature stages ofLiriomyza Trifolii     

Shoshana Yathom  R. Padova  Susana Tal  I. Ross 《Phytoparasitica》1990,18(2):117-124

The effects of γ-radiation on the immature stages ofLiriomyza trifolii Burgess were studied with doses of 2000-25 Gy. The high doses were lethal to all stages. At doses of 750 Gy or less, larvae survived to pupate, but their number was reduced and the majority were either dead or deformed and did not give rise to adults. Eggs and prepupae were more susceptible to radiation than other stages. Larval radiosensitivity decreased with progressing instar, and pupal sensitivity decreased with increasing age. Contribution from the Agricultural Research Organization. No. 2922-E, 1990 series.  相似文献   

Gene discovery in Eimeria tenella by immunoscreening cDNA expression libraries of sporozoites and schizonts with chicken intestinal antibodies   总被引：4，自引：0，他引：4  

Réfega S  Girard-Misguich F  Bourdieu C  Péry P  Labbé M 《Veterinary parasitology》2003,113(1):19-33

Specific antibodies were produced ex vivo from intestinal culture of Eimeria tenella infected chickens. The specificity of these intestinal antibodies was tested against different parasite stages. These antibodies were used to immunoscreen first generation schizont and sporozoite cDNA libraries permitting the identification of new E. tenella antigens. We obtained a total of 119 cDNA clones which were subjected to sequence analysis. The sequences coding for the proteins inducing local immune responses were compared with nucleotide or protein databases and with expressed sequence tags (ESTs) databases. We identified new Eimeria genes coding for heat shock proteins, a ribosomal protein, a pyruvate kinase and a pyridoxine kinase. Specific features of other sequences are discussed.  相似文献   

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis     

Canada N  Meireles CS  Rocha A  Sousa S  Thompson G  Dubey JP  Romand S  Thulliez P  Correia da Costa JM 《Veterinary parasitology》2002,110(1-2):11-15

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.  相似文献   

The dehesa: an agrosilvopastoral system of the Mediterranean region with special reference to the Sierra Morena area of Spain     

Richard Joffre  Jean Vacher  Carlos de los Llanos  Gilbert Long 《Agroforestry Systems》1988,6(1):71-96

A multidisciplinary team undertook a six-year investigation on the agro-ecological and socio-economic aspects of the dehesa system in the Sierra Norte area, a part of the Sierra Morena of the Sevilla Province, Western Andalusia, Spain. This paper summarizes its findings on the various agro-ecological features of the system and evaluates the system's functional aspects vis-a-vis the trends and developments in land use in the region over the past few centuries. The synergistic effects of tree cover on understorey grassland vegetation is discussed in terms of soil fertility build-up and favourable micro-climatic and hydrological features, and the potential use of this information in future research and development programmes to improve the dehesa system in the Mediterranean region is outlined.  相似文献   

Structural ripening-related changes of the arabinan-rich pectic polysaccharides from olive pulp cell walls   总被引：2，自引：0，他引：2  

Cardoso SM  Ferreira JA  Mafra I  Silva AM  Coimbra MA 《Journal of agricultural and food chemistry》2007,55(17):7124-7130

In this study, the structural features and ripening-related changes that occur in the arabinan-rich pectic polysaccharides highly enmeshed in the cellulosic matrix of the olive pulp fruit were evaluated. These pectic polysaccharides, obtained from two consecutive harvests at green, cherry, and black ripening stages, account for 11-19% of the total pectic polysaccharides found in the olive pulp cell walls and were previously shown to occur as calcium chelating dimers. On the basis of the 13C NMR, (1H, 13C) gHSQC, 2D COSYPR, and (1H,13C) gHMBC carbon and proton resonances of the variously linked arabinosyl residues, we propose a tentative structure. This structure is particularly characterized by T-beta-Araf (1-->5)-linked to (1-->3,5)-Araf residues and by the occurrence of branched and linear blocks in the arabinan backbone. Methylation analysis showed that these pectic polysaccharides of black olives have more arabinan side chains, which were shorter (less (1-->5)-Araf), highly branched (more (1-->3,5)-Araf), and with shorter side chains (fewer (1-->3)-Araf) than those of green and cherry olives. Quantitative 13C NMR data indicated that these modifications involved the disappearance of the characteristic terminally linked beta-Araf residue of the arabinans. This odd feature can be used as a diagnostic tool in the evaluation of the stage of ripening of this fruit, as well as a marker for the presence of olive pulp in matrices containing pectic polysaccharides samples.  相似文献   

Inhibition of trypsin by condensed tannins and wine     

Gonçalves R  Soares S  Mateus N  de Freitas V 《Journal of agricultural and food chemistry》2007,55(18):7596-7601

Phenolic compounds are abundant vegetable secondary metabolites in the human diet. The ability of procyanidin oligomers and wine polyphenols to inhibit trypsin activity was studied using a versatile and reliable in vitro method. The hydrolysis of the chromogenic substrate N-benzoyl-d,l-arginine-p-nitroanilide (BApNA) by trypsin was followed by spectrophotometry in the presence and absence of condensed tannins and wine. A clear relationship between the degree of polymerization of procyanidins and enzymatic inhibition was observed. Trypsin activity inhibition was also detected in several types of wine. In general, the inhibition increased with the concentration of phenolic compounds in wines. These results may be relevant when considering these compounds as antinutritional factors, thereby contributing to a reduced absorption of nutrients.  相似文献   

Cloning of two Atlantic salmon growth hormone receptor isoforms and <Emphasis Type="Italic">in vitro</Emphasis> ligand-binding response     

Susana?BenedetEmail author  Viktoria?Johansson  Glen?Sweeney  Malyka?Galay-Burgos  Bj?rn?Thrandur?Bj?rnsson 《Fish physiology and biochemistry》2005,31(4):315-329

Two isoforms of the full-length cDNA of the growth hormone receptor (GHR) of the Atlantic salmon (Salmo salar; ss) were cloned by a PCR approach using RACE. Respectively, the cDNA sequences of ssGHR isoforms 1 and 2 are 2654 and 2608 nucleotides long, with 1782 and 1773 nucleotide ORFs. The resulting coded proteins are 594 and 590 aa long, with 19 and 20 aa signal peptides. The two isoforms share 86% protein and 87% cDNA sequence similarity. Isoform 1 is most similar to other salmonid GHR isoforms 1 while isoform 2 is most similar to salmonid GHR isoforms 2 (93–95%). Similarity with other teleost species was lower (37–44%). The bioactivity of the cloned ssGHR was tested by transfecting the ssGHR isoform 1 cDNA into CHO-K1 hamster cells, incubating with recombinant salmon GH (sGH) or native ovine prolactin (oPRL), and measuring cell proliferation by the MTT assay. The ssGHR-transfected cells significantly increased proliferation when stimulated by sGH at all concentrations. oPRL stimulated ssGHR-transfected cells at higher concentrations due to receptor cross reaction. ssGHR isoforms 1 and 2 contain a single transmembrane domain and the typical conserved motifs found in other teleost GHRs, including four paired cysteine residues and five potential N-glycosylation sites in the extracellular domain, Box I and Box II, as well as seven potential tyrosine phosphorylation sites in the intracellular domain. However, in salmonids, these motifs differ from those of other teleosts, and could be responsible for differentiated hormone binding, signal transduction and response.  相似文献   

Isolation,in vitro propagation,genetic analysis,and immunogenic characterization of an Ehrlichia canis strain from southeastern Brazil     

Rosiane Nascimento Alves  Susana Elisa Rieck  Carlos Ueira-Vieira  Marcelo Bahia Labruna  Marcelo Emílio Beletti 《Journal of veterinary science (Suw?n-si, Korea)》2014,15(2):241-248

Amplification of the 16S rRNA gene from a blood sample obtained from a dog in southeastern Brazil was used to confirm a naturally acquired Ehrlichia (E.) canis infection. Following isolation and culturing of the new bacterial strain called Uberlândia, partial sequences of the dsb and p28 genes were obtained. The dsb partial sequence of the novel strain was 100% similar to dsb gene sequences of E. canis obtained from different geographic areas around the world. Conversely, the p28 partial sequence for the E. canis Uberlândia strain differed at several nucleotides from other sequences available in GenBank. To confirm the antigenic profile of the Uberlândia strain, an indirect immunofluorescence assay against E. canis antigens was performed using dog sera collected from two different areas in Brazil (Uberlândia and São Paulo). The results suggest that both antigens were able to identify animals seropositive for E. canis in Brazil since these Brazilian strains appear to be highly conserved.  相似文献