Protease Inhibitors from Marine Venomous Animals and Their Counterparts in Terrestrial Venomous Animals

The Kunitz-type protease inhibitors are the best-characterized family of serine protease inhibitors, probably due to their abundance in several organisms. These inhibitors consist of a chain of ~60 amino acid residues stabilized by three disulfide bridges, and was first observed in the bovine pancreatic trypsin inhibitor (BPTI)-like protease inhibitors, which strongly inhibit trypsin and chymotrypsin. In this review we present the protease inhibitors (PIs) described to date from marine venomous animals, such as from sea anemone extracts and Conus venom, as well as their counterparts in terrestrial venomous animals, such as snakes, scorpions, spiders, Anurans, and Hymenopterans. More emphasis was given to the Kunitz-type inhibitors, once they are found in all these organisms. Their biological sources, specificity against different proteases, and other molecular blanks (being also K⁺ channel blockers) are presented, followed by their molecular diversity. Whereas sea anemone, snakes and other venomous animals present mainly Kunitz-type inhibitors, PIs from Anurans present the major variety in structure length and number of Cys residues, with at least six distinguishable classes. A representative alignment of PIs from these venomous animals shows that, despite eventual differences in Cys assignment, the key-residues for the protease inhibitory activity in all of them occupy similar positions in primary sequence. The key-residues for the K⁺ channel blocking activity was also compared.     

Evaluation of survival of Actinobacillus pleuropneumoniae and Haemophilus parasuis in four liquid media and two swab specimen transport systems

OBJECTIVE: To determine duration and rates of recovery of Actinobacillus pleuropneumoniae and Haemophilus parasuis from 4 liquid media and 2 swab specimen transport systems and compare findings with those of Escherichia coli. SAMPLE POPULATION: One strain each of A pleuropneumoniae (biovar 1, serotype 1), H parasuis (serovar 5), and E coli (serotype O149:K91:H19). PROCEDURE: Strains were incubated in brain heart infusion broth supplemented with horse serum and other nutrients or in horse serum alone, with and without nicotinamide-adenine dinucleotide in both instances, for 150 days at 4 degrees C or room temperature (21 degrees C). Similarly, strains were tested in Stuart and Amies transport systems after storage at room temperature for 8 days. RESULTS: Colony counts greater than those of the initial inoculum were observed after incubation in horse serum for A pleuropneumoniae but not for H parasuis. Overall, incubation at 4 degrees C in the 4 liquid media resulted in longer recovery duration and higher rates than at room temperature. Culture of H parasuis resulted in lower recovery rates and shorter durations of recovery than culture of A pleuropneumoniae, except for culture in horse serum. Haemophilus parasuis survived longer than A pleuropneumoniae in the transport systems, and all organisms survived longer in the Amies system. CONCLUSIONS AND CLINICAL RELEVANCE: Survival of A pleuropneumoniae and H parasuis indicated that horse serum prolongs survivability, which may result in exposure of more animals during a prolonged period. The Amies system might be a good choice for collection of clinical samples from animals, especially for recovery of H parasuis.     

Effects of month of breeding on reproductive efficiency of Holstein cows and heifers inseminated with sex-sorted or conventional semen in a hot environment

The main objective of this study was to assess the effect of month of breeding on reproduction performance of Holstein heifers and cows inseminated with sex-sorted or conventional semen in a hot environment. Pregnancy per artificial insemination (P/AI; 64,666 services over an 8-year period) both in heifers (n?=?22,313) and cows (n?=?42,353) from a large dairy herd in northern Mexico (26°N) were evaluated with the GENMOD procedure of SAS, with respect to month of AI. Overall, P/AI with sex-sorted semen was greater (P?<?0.01) in heifers (41.6 %) than cows (17.3 %). P/AI for cows serviced with conventional semen was 10 % points higher (P?<?0.01) in January and December (31 vs. 21 %) than cows serviced with sex-sorted semen. While there was no difference in P/AI between the sex-sorted sperm and conventional semen in cows inseminated in July (16 and 18 %, respectively), P/AI plummeted for both groups of cows during the summer and fall (more severe heat stress). P/AI was not different between heifers serviced with sex-sorted or conventional semen during the hottest months of the year (July to October). However, during the coldest month of the year (January and February), P/AI was 10 percentage points greater (P?<?0.01) in heifers serviced with conventional than sex-sorted semen. It was concluded that in this hot climate cow and heifer fertility declined in the summer and fall when inseminated with conventional semen. However, the use of sex-sorted semen during summer and fall did not compromise the breeding success in heifers. Thus, this data suggest that sex-sorted semen promotes some embryonic thermoprotective mechanism, which leads to a marginal summer and fall fertility depression with this type of semen in this particular hot environment.     

Performance, growth, and maturity of Nellore bulls

The objectives of this study were to evaluate the dry matter intake (DMI), digestibility, average daily gain (ADG), microbial efficiency, empty body weight (EBW) gain, and body composition of Nellore bulls. Additionally, Nellore bull maturity was estimated, and the prediction equation for DMI, suggested by the Brazilian nutrient requirements system (BR CORTE; Azevêdo et al. 2010), was evaluated. Thirty-three Nellore bulls, with a mean initial weight of 259?±?25 kg and age of 14?±?1 months, were used in this study. Five animals were slaughtered at the beginning of the experiment (control group), and the remaining 28 were divided into 4 groups, each slaughtered at 42-day intervals. Their diet was composed of corn silage and concentrate (55:45). The power model was used to estimate muscle tissue, bone tissue, crude protein (CP), mineral matter (MM), and water present in the empty body, while the exponential model was used to estimate adipose tissue and ether extract (EE) present in the empty body. When expressed in kilograms per day, differences were observed (P?<?0.05) only for the intake of EE and neutral detergent fiber as a function of feedlot time periods. Although there was a difference in relation to nutrient intake, it did not affect (P?>?0.05) digestibility, with the exception of EE digestibility. The equation suggested by BR CORTE correctly estimates the DMI of Nellore bulls. ADG was not affected (P?>?0.05) by time spent in the feedlot. No differences were observed (P?>?0.05) for microbial efficiency; a mean value of 142 g microbial crude protein/kg total digestible nutrients was achieved. The muscle and bone tissues, CP, MM, and water present in the empty body increased as the animal grew, although at a lower rate. The adipose tissue and EE present in the empty body increased their deposition rate when the animal reached its mature weight. Maturity is defined as when an animal reaches 22 % EE in the empty body, which corresponds to 456 kg of EBW in Nellore bulls. Therefore, this study can conclude that the feedlot time period does not affect DMI, nutrient intake, ADG, or microbial efficiency. The equation proposed by BR CORTE (Azevêdo et al. 2010) correctly estimates the DMI of Nellore bulls, which reach maturity when an EBW of 456 kg is attained.     

Ae.ovata细胞质小麦的核质杂种优势研究

利用恢复系 R113与10种不同细胞质的异质小麦雄性不育系 Selkirk(或Chris)杂交,以鉴定 R113对这些雄性不育系的恢复力。其中有7种细胞质的雄性不育性在不同程度上为 R113所恢复。证明了 R113恢复性的广泛性。特别值得注意的是 R113对 Ae.ovata 细胞质不育的恢复度极高,其恢复性可能由两对恢复基因控制.根据杂交种与其亲本之间以及同质 Selkirk 与异质 Selkirk 品种之间的比较,大多数所测性状,如株高、抗倒伏性、每穗小穗数、每穗粒数、每穗粒重等,在 Ae.ovata 细胞质背景下都有明显的优势.试验证明,在小麦中确实存在着两种杂种优势:即核间杂种优势和核质间杂种优势。Ae.ovata 细胞质的缺点是较晚熟,然而,延长春化时间可以提早抽穗;在其同早熟亲本杂交的后代中,也可以选出早熟植株.在小麦常规育种中或在杂种优势的利用上,可以应用 Ae.orata 细胞质把上述两种杂种优势结合起来,选育优良的小麦核质杂种品种或杂种小麦。     

Effect of salinity on metalloenzymes of oxygen metabolism in two leguminous plants

Abstract

Superoxide dismutase (SOD) pattern, catalase, Cyt c oxidase and fumarase activity were studied in leaves of Phaseolus vulgaris and Vigna unguiculata plants growth in two sodium chloride (NaCl) concentrations (35 mM and 100 mM). In bean plants growth with NaCl, leaf chloride (Cl^?) contents were higher than in control plants, and the same was found for sodium (Na⁺) and potassium (K⁺) contents, although to a lesser degree. In cowpea leaves, Na⁺ and Cl^? had a similar increase due to salt‐growth conditions. Under salinity, all changes in the antioxidant (SOD and catalase) enzymes levels were smaller in bean than in cowpea plants. In Phaseolus at 15 days growth, Cu, Zn‐SOD I showed an increase by the effect of salt treatment, but this induction did not occur at 30 days growth, and both Mn‐SOD and Cu, Zn‐SOD II did not show variations due to salt‐stress. In Vigna, Mn‐SOD was decreased by salinity but this was compensated by an increase in Cu, Zn‐SOD I activity in plants at 30 days growth, whereas in young leaves under saline conditions, both isozymes were also decreased. Likewise, there was a rise in cytochrome c oxidase and fumarase activity in leaves of NaCl‐treated plants compared to the control. The activity changes observed are discused in term of their possible relevance to plant sensitivity to saline conditions.     

Further studies on the analysis of DSP toxin profiles in galician mussels

Further studies on mussel samples from Galicia, Spain, have revealed the presence of okadaic acid (OA), dinophysistoxin 2 (DTX2), and the fatty acid acyl esters of both of these toxins as the "DTX3" complex. Measurements were performed with an improved in situ method for the formation of 9-anthryldiazomethane (ADAM) derivatives followed by liquid chromatography with fluorescence detection. Base hydrolysis of DTX3 toxins gave free OA and DTX2, which were determined following ADAM derivatization. Results were confirmed by liquid chromatography/mass spectrometry analyses, and in most of the samples, free DTX2 was the most abundant toxin. However, the OA/DTX2 ratio in the DTX3 conjugated form was different, with OA being the most abundant in all cases. This difference could be due to different rates of metabolism of OA and DTX2 to the acyl esters or due to contamination of the shellfish by the two toxins at different points in time, resulting in less acyl ester formation for one toxin versus the other. The second possibility would be reasonable if two different source organisms were producing the toxins.     

Clinical findings related to intramammary infections in meat-producing ewes

The aim of this study was to investigate the relationship between clinical findings and bacterial isolation in milk samples of meat-producing ewes. The study was conducted in 17 commercial flocks and 550 udder halves from suckling Santa Ines ewes. Initially, the clinical examination of the mammary glands and teats was performed by visual inspection and palpation of the teats and udder halves; then a scoring system was devised for all the findings. After that, the strip cup test and the California mastitis test (CMT) were performed. Then, milk samples for somatic cell counts (SCCs) and bacteriological analyses were collected. Staphylococci bacteria were the main etiological agent isolated in the present study. Upon investigation of the correlations between bacterial isolation and the clinical findings, only the presence of teat injury, pendulous udder, and alterations in the palpation of the teat were associated with bacterial isolation. A significant correlation between bacteriologically positive milk samples and CMT and SCC was also found. Thus, some clinical findings appeared as a risk factor for bacteriologically positive milk samples and can be used as a tool in mastitis control programs. However, a complete and extensive diagnosis, an appropriate therapy, and an efficient mastitis control program will require the combination of clinical examination, microbiological tests, and SCC.