The Effect of Molecular Weight on the Antimicrobial Activity of Chitosan from Loligo opalescens for Food Packaging Applications

The growing requirement for sustainable processes has boosted the development of biodegradable plastic-based materials incorporating bioactive compounds obtained from waste, adding value to these products. Chitosan (Ch) is a biopolymer that can be obtained by deacetylation of chitin (found abundantly in waste from the fishery industry) and has valuable properties such as biocompatibility, biodegradability, antimicrobial activity, and easy film-forming ability. This study aimed to produce and characterize poly(lactic acid) (PLA) surfaces coated with β-chitosan and β-chitooligosaccharides from a Loligo opalescens pen with different molecular weights for application in the food industry. The PLA films with native and depolymerized Ch were functionalized through plasma oxygen treatment followed by dip-coating, and their physicochemical properties were assessed by Fourier-transform infrared spectroscopy, X-ray diffraction, water contact angle, and scanning electron microscopy. Their antimicrobial properties were assessed against Escherichia coli and Pseudomonas putida, where Ch-based surfaces reduced the number of biofilm viable, viable but nonculturable, and culturable cells by up to 73%, 74%, and 87%, respectively, compared to PLA. Biofilm growth inhibition was confirmed by confocal laser scanning microscopy. Results suggest that Ch films of higher molecular weight had higher antibiofilm activity under the food storage conditions mimicked in this work, contributing simultaneously to the reuse of marine waste.     

Feminization and growth of juvenile fat snook Centropomus parallelus fed diets with different concentrations of estradiol-17β

In this study, we evaluated the effect of diets with various concentrations of estradiol-17β (E₂) on the feminization, growth and survival of juvenile fat snook. Over a 45-day period, the juveniles were fed with five diets containing 0, 25, 50, 75 and 100 mg E₂/kg feed. Three replicates were run for each diet. After this hormonal treatment, the fishes were transferred to 1-m³ net cages, where they remained for 12 months. During this period, they were fed a commercial diet without added hormones. During the feminization, the control fish grew more than those in the other treatments, and the survival rates did not differ significantly among the treatments. At the end of the experiment growth did not differ among the treatments. In the control treatment, 80.85 % of the fish were male, and the females were significantly larger than the males. All of the treatments with E₂ produced 100 % females. No intersex fish were found in this study. This research showed that it is possible to obtain 100 % fat snook females using feeds with 25 mg E₂/kg for 45 days without impairing the growth or survival of the fish.     

Habituation and conditioning in gilthead sea bream (Sparus aurata): Effects of aversive stimuli,reward and social hierarchies

To tailor the farming environment to a fish species, we should understand the species‐specific responses to stimuli, including the degree of adaption and learning. Groups of gilthead sea bream were given a delay Pavlovian conditioning regime using a conditioning stimulus (CS) of light flashes signalling arrival of food. Controls were exposed to light flashes unrelated to feeding. Fish in both treatments showed an initial fear response of moving away from the CS combined with reduced swimming speed. In subsequent trials, the Control fish largely habituated the fleeing response but sustained to respond by reducing the swimming speed. The Conditioning fish also stopped to escape from the CS, but opposed to the Control group they gradually increased their swimming speed in response to the CS. In addition, the number of fish in the feeding/CS area increased and became similar to basal level after around 16 trials. A small and variable proportion of the fish displayed black vertical bands on their body and territorial behaviour, and a social hierarchy could interfere with the processes of habituation and conditioning. The swimming speed of the fish increased with number of dark individuals, but this was not found during the CS and the light stimulus thus seemed to overrule the effect of territorial behaviour. The persistent negative response to light flashes in the Control suggests that fish seemingly adapted to repetitive stressors are still in a state of alertness. The change in the response to light shows the potential for rewarding aversive stimuli to reduce stress.     

Serotypes and Shiga toxin genotypes among Escherichia coli isolated from animals and food in Argentina and Brazil

Shiga toxin (Stx)-producing Escherichia coli (STEC) strains isolated from animals and food in Argentina (n=44) and Brazil (n=20) were examined and compared in regard to their phenotypic and genotypic characteristics to evaluate their pathogenic potential. The clonal relatedness of STEC O157 isolates (n=22) was established by phage typing (PT) and pulsed-field gel electrophoresis (PFGE). All O157 strains studied carried eae and enterohemorrhagic E. coli (EHEC)-hly sequences. In Argentina, these strains occurred both in cattle and meat, and 50% of them carried stx2/stx2vh-a genes, whereas in Brazil the O157 strains were isolated from animals, and most harbored the stx2vh-a sequence. At least 13 different O:H serotypes were identified among the non-O157 strains studied, with serotype O113:H21 being found in both countries. All but one non-O157 strains did not carry eae gene, but EHEC-hlyA gene was found in 85.7% of them, and the stx2 genotype was also more prevalent in Argentina than in Brazil (P<0.01), where stx1 alone or in association was most common (68.8%). One STEC strain isolated from a calf in Brazil harbored the new variant referred to as stx2-NV206. PFGE analysis showed that STEC O157 strains were grouped in four clusters. One Brazilian strain was considered possibly related (> or =80%) to Argentinean strains of cluster I. Differences in the pathogenic potential, especially in regard to serotypes and stx genotypes, were observed among the STEC strains recovered from animals and food in both countries.     

Spray‐dried porcine plasma improves feed intake of weaned piglets subjected to heat stress

The objective of this study was to evaluate the effects of spray‐dried porcine plasma (SDPP) on performance, measurement of thyroid hormones triiodothyronine (T3) and thyroxine (T4), body temperature and relative organ weights of weaned piglets exposed to a heat stress or a thermoneutral environment. One hundred and forty‐four piglets with initial BW of 5.82 ± 0.33 kg (28 d old) were used. The experimental design was a 4 x 2 factorial arrangement, four plasma levels (0.0, 2.5, 5.0 and 7.5%) and two environments (34º [heat stress (HS)] and 24°C [thermoneutral (TN)]). SDPP was fed from weaning (28 d of age) to the end of the trial (piglets were weighed at the beginning, 7, 21 and 35 days of trial). Rectal, neck, shoulder and leg temperatures were measured every seven days. At 21 days of trial, one piglet per pen was slaughtered to determine the weight of the carcass and organs. Blood samples were collected at 18 and 35 days of trial to determine levels T3 and T4. The rectal, neck, shoulder and leg temperatures were increased (p < 0.05) 0.4, 1.5, 1.0 and 1.3ºC, respectively, and relative weight of the lungs (p < 0.05) and total T3 (p < 0.05) were decreased 0.001 kg/kg and 0.19 ng/dl, respectively, for pigs in the HS environment. The estimated inclusion of 5.1% of SDPP had the greatest (p < 0.05) weight gain during the first week, and throughout the nursery phase, the 5% of inclusion had a tendency of improvement (p = 0.075), regardless of ambient temperature. For the entire period, pigs reared in the HS environment had the greatest (p < 0.05) estimated feed intake by the quadratic regression with the inclusion of 4.3% of SDPP.     

An enzyme-linked immunosorbent assay (ELISA) test for the diagnosis of equine infectious anemia in Brazil

An enzyme-linked immunosorbent assay (ELISA) test was developed for the detection of specific antibodies against the unique infectious anemia (EIA) virus in equine sera. The ELISA test was faster and more sensitive when compared with the classic test of agar gel immunodiffusion (AGID). A total of 200 sera were tested: 100 from negative horses and 100 from positive horses by AGID. The ELISA test showed 92 horse sera negative and 100 horse sera positive by AGID with values of optical density (OD) less than 0.139 and higher than 0.139, respectively. Eight horse sera were negative by AGID and higher than 0.139 by ELISA. Six of these became AGID positive also when re-tested 30 days later, and two were of the horses that showed clinical signs of EIA and died before re-testing.     

Virulence markers and genetic relationships of Shiga toxin-producing Escherichia coli strains from serogroup O111 isolated from cattle

Shiga toxin-producing Escherichia coli (STEC) strains isolated from healthy cattle (O111:NM, seven strains; O111:H8, three strains) in Brazil were studied and compared to previously characterized human strains in regard to their phenotypic and genotypic characteristics to evaluate their pathogenic potential. Most bovine STEC O111 strains were isolated from dairy calves, and strains with genotypes stx1 alone and stx1/stx2 (variant stx2) occurred in different regions. Irrespective of the stx genotype, all strains were positive for eae theta, alpha variants of tir, espA and espB, and for ler, qseA, iha, astA and efa1 genes. Only one strain was negative for EHEC-hlyA and all strains were negative for iha, saa and espP genes and for EAF and bfpA, genetic markers of EPEC. Except for the presence of stx2, bovine strains showed the same profile of putative virulence genes found among the human strains. Similar biochemical behavior was identified among the strains analysed. Two bovine STEC strains produced the localized adherence (LA) phenotype in 6-h tests with Caco-2 (human enterocyte) cells. Intimate attachment (judged by the FAS test) was found in 9 out of 10 bovine strains as it was observed for the human STEC strains. RAPD-PCR analysis showed two distinct RAPD groups among the STEC O111 strains examined. Despite the relative low frequency of STEC O111 strains recovered from cattle no differences in their pathogenic potential were observed compared to some strains isolated from human diarrhea, suggesting that healthy cattle may be a potential source of infection for humans in Brazil.     

Land use vs. fragment size and isolation as determinants of small mammal composition and richness in Atlantic Forest remnants

The remaining Atlantic Forest fragments are structurally isolated by a matrix of pastures, plantations, or urban areas, and most remnants are small (<100 ha). Island biogeography theory has been used to predict the effects of such fragmentation in the remaining fragments, but human activities and land use around fragments may be equally important. A related question is which aspects of land use have a strong effect on biodiversity. We compare the relative importance of fragment size and isolation vs. land use around fragments as determinants of composition and richness of small mammals in Atlantic Forest fragments. We also compare two aspects of land use around fragments, economic activity (peri-urban, agriculture, cattle), and property ownership (peri-urban, low income rural producers, affluent rural producers). Small mammals were surveyed in 21 fragments varying from 12 to 250 ha, and in two sites of continuous forest in the Macacu River watershed, State of Rio de Janeiro, Brazil, from 1999 to 2007. The effects of land use, fragment size and isolation were formulated as eleven candidate models, compared by Akaike Information Criteria. In the models selected, species composition was associated more strongly with fragment size, followed by isolation, with a smaller effect of property ownership. Species richness was determined mostly by fragment isolation, but also by a negative effect of agriculture when it was the dominant economic activity. Regardless of the critics to island biogeography theory, fragment isolation and size were by far the most important determinants of species composition. Economic activity and property ownership allowed the detection of subtle but important effects of land use on species composition and richness.