Identification and Characterisation of Bacteria Causing Soft-rot in Agave tequilana

Agave tequilana is the raw material for the production of the alcoholic beverage tequila. A bacterial disease has affected the A. tequilana crop in recent years. Previous reports based on colony and cell morphology, Gram stain and potato rot indicated that Erwinia sp. is the main pathogen. We isolated a several bacterial isolates capable of producing soft-rot symptoms in greenhouse pathogenicity assays. An extensive characterisation involving pathogenicity tests, fatty acid profile, metabolic and physiological properties, ribosomal DNA sequence and intergenic transcribed spacer amplification (ITS-PCR) and restriction banding pattern (ITS-RFLP) was made of each isolate. Three different species: Erwinia cacticida, Pantoea agglomerans and Pseudomonas sp. were identified. Fatty acid and metabolic profiles gave low similarity values of identification but 16S rDNA sequence, ITS-PCR and ITS-RFLP confirmed the identification of E. cacticida. In the phylogenetic tree, E. cacticida from blue agave was grouped neither with E. cacticida type strains nor with Erwinia carotovora. This is the first report that associates E. cacticida with A. tequilana soft-rot symptoms.     

Comparative Characterization of Enzymatic Digestion from Fish and Soybean Meal from Simulated Digestive Process of Pacific Bluefin Tuna,Thunnus orientalis

The digestive process of the Pacific bluefin tuna (PBT), Thunnus orientalis, was simulated through two phases of in vitro digestion: acidic digestion with porcine pepsin, followed by alkaline digestion with pancreatic crude extract (PCE) obtained from the PBT to hydrolyze fish meal (FM) and soybean meal (SBM) as protein substrates. The crude protein from FM resulted in a lower degree of hydrolysis (73.3%) compared with SBM (79.2%). However, the resulting digested products showed that FM contained 35% more small peptides, with sizes <6.5 kDa than those from the starting material (>150 kDa). The SBM had an increase of only 1.3% in the similar peptide cut‐offs found after hydrolysis. These results suggested that FM appeared to be a better source of protein according to the amount of low‐molecular weight peptides. In addition, the proteolytic activity of PCE showed that 88.9% of its alkaline proteolytic activity corresponded to trypsin and 2.9% corresponded to chymotrypsin activity. The results shown here demonstrate that peptide sizes are important in identifying suitable protein sources for aquafeed production to reinforce the primary results obtained from the in vitro digestibility using the pH‐Stat system. These results also contribute to a better understanding of the digestibility process in aquatic organisms.     

Survival and Growth Improvement of Palm Ruff,Seriolella violacea,Larvae Fed Artemia Nauplii Enriched with an Experimental Emulsion

The palm ruff, Seriolella violacea (Cojinoba), is a potential new species for Chilean aquaculture. To approach Cojinoba larviculture, an experimental Artemia enrichment emulsion, containing docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) = 2.5, supplemented with vitamin E, astaxanthin, and β‐glucan, was evaluated in both Artemia and Cojinoba larvae, 30–50 d.a.h. This study tested an experimental enrichment emulsion versus a commercial emulsion, with an integral approach of multicompound emulsions. After 23 h enrichment, experimental emulsion (EE)‐enriched nauplii reached DHA and EPA concentrations of 23.8 and 18.7 mg/g dry weight (dwt), respectively, while in Cojinoba larvae they were 18.4 and 19.7 mg/g dwt. Control emulsion (CE)‐enriched nauplii exhibited lower DHA and EPA (6.1 and 7.7 mg/g dwt), while only DHA decreased in the control larvae (12.6 mg/g dwt). Vitamin E was higher in EE‐enriched nauplii (29.2 mg/100 g dwt) than in the control (8.4 mg/100 g dwt). Larvae fed EE‐enriched Artemia exhibited 8% increase in survival and 19% in growth compared with the control. Astaxanthin was detected only in larvae fed EE‐enriched nauplii. The tumor necrosis factor‐α concentration was not significantly different between larvae fed EE‐ and CE‐enriched nauplii. EE looks promising as an Artemia enrichment and experimental diet to assess palm ruff larval requirements, and has a positive impact on fish larvae performance.     

Anatomical Variations of the Blood Vascular System in Veterinary Medicine. The Internal Iliac Artery of the Dog. Part Two

The aim of this study was to investigate the variability of the internal pudendal artery. Two hundred and thirty‐two pelvic halves from 116 adult dogs were examined. Twenty‐six anatomical variations were found, thirteen occurring in more than 5% of the dogs. Anatomical variations were grouped in relation to the origin of the prostatic/vaginal arteries, middle rectal artery, urethral artery, ventral perineal and caudal rectal arteries. The chi‐squared test was used to analyse differences in sex, side of the body, profile and size, and the results were considered statistically significant when P ≤ 0.05. An identical vascular pattern in both hemipelvises was found for most of the anatomical variations described.     

Anatomical Variations of the Blood Vascular System in Veterinary Medicine: The Internal Iliac Artery of the Dog — Part Three

The aim of this study was to analyse and describe the variability of the umbilical artery. Two hundred and thirty‐two pelvic halves from 116 adult dogs were examined. To study the permeability of the umbilical artery, ten adult dogs, nine newborns and thirteen foetuses between 35 and 50 days of gestation were also used. In relation to the origin of the umbilical artery, six anatomical variations were found. From which five involved a cranial (n = 4) or caudal (n = 1) relocation of its origin, and in one case (n = 1), the umbilical artery arose from the median sacral artery. In eight cases, the umbilical artery gave off the prostatic (n = 1) or vaginal (n = 7) arteries. The permeability of the umbilical artery was the most significant anatomical variation: permeability was detected in 45% (106 of 232 pelvic halves) of all cases, from which 36 were males and 70 females. Interestingly, an equal vascular permeability in both hemipelvises was found for 82% of the dogs, thus additional data related to such feature of the umbilical artery was also recorded. In accordance with the statistical study, the main anatomical variations described showed significant values for gender, side of the body, size and profile variables.     

Adrenomedullin (AM) and adrenomedullin binding protein (AM-BP) in the bovine mammary gland and milk: Effects of stage of lactation and experimental intramammary E. coli infection

Adrenomedullin (AM) has been characterized as an endogenous tissue survival factor and modulator of many inflammatory processes. Because of the increased susceptibility of the mammary gland to infection during the time surrounding parturition in the cow, we investigated how milk and tissue content of AM and its binding protein (AM-BP) might be affected by the stage of lactation and the udder health status. Milk and mammary biopsy samples were obtained from Holstein cows 21 days prior to and at various times after calving to represent the dry period and early and mid-stages of lactation. Additional cows received an intramammary challenge with Escherichia coli for immunohistochemical characterization of AM and AM-BP. Milk AM concentrations were relatively constant across the stages of lactation while AM-BP increased two-fold (P<0.04) between early and mid-lactation. Milk AM (P<0.04) and AM-BP (P<0.03) increased as somatic cell counts (SCCs) increased within a given stage of lactation. Tissue content of both (AM and AM-BP) were significantly affected by stage of lactation, lowest in the dry period and progressively increasing to peak at mid-lactation as well as increasing in association with higher levels of SCCs. Following E. coli challenge, AM increased in epithelial cells surrounding mammary alveoli presenting high levels of SCCs. The data suggest that AM and AM-BP are cooperatively regulated in the mammary gland during lactation; changes in localized tissue AM and AM-BP content reflect a dynamic regulation of these tissue factors in the bovine mammary gland consistent with their protective effects within inflamed tissue.