Optimization of multiple headspace solid-phase microextraction for the quantification of volatile compounds in dry fermented sausages

Multiple headspace solid-phase microextraction (HS-SPME) is a stepwise method that eliminates the influence of the matrix sample on the quantitative analysis of solid samples. The process was optimized for the analysis of volatile compounds in dry fermented sausages by gas chromatography and mass spectrometry. Different amounts of fermented sausages and different vial volumes were studied to obtain the theoretical exponential decay of the peak area of the four successive extractions in order to calculate the total area in the sausage. The highest number of volatile compounds analyzed by multiple HS-SPME in dry fermented sausages was obtained in a 10 mL headspace vial with 0.07 g of sample in the presence of water, 0.75 mg butylated hydroxytoluene, and 0.5 g sodium chloride. Finally, the method was characterized in terms of linearity and detection limits and applied to analyze the volatile compounds present in fermented sausages manufactured with either nitrate or nitrite.     

Broccoli processing wastes as a source of peroxidase

A peroxidase isozyme (BP) was purified to homogeneity from broccoli stems ( Brassica oleraceae var. maraton) discarded from industrial processing wastes. BP specific activity was 1216 ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)] units/mg, representing 466-fold that of crude extract. BP is a monomeric glycoprotein containing 16% carbohydrates, with a molecular mass of 49 kDa and an isoelectric point close to 4.2. From kinetic data it showed a two-substrate ping-pong mechanism, and the catalytic efficiency measured as the rate-limiting step of free BP regeneration was 3.4 x 10(6) M(-1) s(-1). The ABTS K m value was 0.2 mM, which was about 20 times lower than that reported for acidic commercial horseradish peroxidase (HRP). Assessment of BP secondary structure showed 30% helical character, similar to HRP and cytochrome c peroxidase. BP lost only 25% activity after 10 min of heating at 55 degrees C and pH 6; it was stable in the pH range from 4 to 9 and showed an optimum pH of 4.6 using ABTS as substrate. BP was active on substrates normally involved in lignin biosynthesis, such as caffeic and ferulic acids, and also displayed good catechol oxidation activity in the presence of hydrogen peroxide. Reverse micellar extraction was successfully used as potential large-scale prepurification of broccoli peroxidase, achieving a purification factor of 7, with 60% activity yield. Stems from the broccoli processing industry have a high potential as an alternative for peroxidase purification.     

Influence of diet and feeding strategy on the performance of nitrifying trickling filter,oxygen consumption and ammonia excretion of gilthead sea bream (Sparus aurata) raised in recirculating aquaculture systems

Gilthead sea bream (Sparus aurata) was raised in six individual recirculating aquaculture systems (RAS) whose biofilters’ performance was analyzed. Fish were fed with three different diets (a control diet, a fishmeal-based diet (FM), and a plant meal-based diet (VM)) and with three different feeding strategies (manual feeding to apparent satiation, automatic feeding with restricted ration, and auto-demand feeding). For every combination of diet and feeding strategy, the mean oxygen consumption, ammonia excretion, and ammonia removal rate were determined. Fish fed with the VM diet consumed the most oxygen (20.06?±?1.80 gO₂ consumed kg^?1 day^?1). There were significant differences in ammonia excretion depending on the protein content and protein efficiency of the diet, as well as depending on feeding strategy, which in turn affected ammonia removal rates. Fish fed by auto-demand feeders led to the highest mean ammonia removal rate (0.10 gN-TAN removed m^?2 biofiltration area day^?1), while not leading to peaks of high ammonia concentration in water, which preserve fish welfare and growth.

     

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.     

Influence of S. babylonica extract on feed intake, growth performance and diet in vitro gas production profile in young lambs

An experiment was completed to determine the effect of Salix babylonica (SB) extract supplementation to the diet of growing lambs. Eighteen Katahdin × Pelibuey male lambs (14?±?2 kg live body weight) were divided randomly in individual cages into three groups and fed three diets varying in SB: a control group was fed on total mixed ration (TMR) without SB (SB0), an SB25 group was fed on TMR plus SB extract at 25 mL/lamb/day, and an SB50 group was fed on TMR plus SB extract at 50 mL/lamb/day on dry matter intake (DMI), average daily gain (ADG), feed efficiency, and in vitro gas production (GP) in lambs fed on TMR. In vitro GP of the TMR fed to lambs was recorded at 2, 4, 6, 8, 10, 12, 24, 48, and 72 h of incubation with 0, 0.6, 1.2, and 1.8 mL extract per gram of DM. Addition of SB extract at low and high doses improved the DMI of lambs by 59.9 and 33.2 %, respectively. Relative to the control, low and high extract doses achieved greater lamb ADG during the experimental period. The asymptotic GP increased (P?<?0.05) with increasing dose of SB extract without affecting the rate of GP or the initial delay before GP begins. Linear increases for in vitro GP with advancing time with different SB extract doses were observed. It is suggested that the use of S. babylonica extract with the rate of 25 mL/lamb/day is beneficial to young lamb’s performance growth and thus can be safely used as a feed additive in diets without any negative effects on animal health.     

Milk yield and reproductive performance of brucellosis-vaccinated but seropositive Holstein cows

The aim of this research was to study if seropositivity for brucellosis in vaccinated cows against this disease hampers reproductive performance and milk production in high-yielding Holstein cows. For this purpose 1,026 healthy cows and 372 cows seropositive for brucellosis were enrolled in this study. Cows positive to card test and subsequently to the rivanol test were further subjected to the radial immunodiffusion (RID) test. It was found that only 11 % of the presumably infected cows by brucellosis screening tests were really infected with this disease. The reproductive performance of the group of cows with 11 % Brucella-infected animals was not impaired; overall pregnancy rate did not differ between seropositive and healthy cows (30.9 vs. 29.6 %). The abortion rates were similar between seropositive cows (5.3 %) and seronegative animals (6.9 %). Cows in the herd with 11 % Brucella-infected animals produced significantly more milk than unaffected cows over a 305-day lactation (10,684?±?1,720 vs. 10,345?±?1,736; mean ± SD; P?<?0.05). It was concluded that in dairy herds vaccinated against brucellosis with both 19 and RB51 strains, supplemental tests such as RID need to be conducted on all reactors in order to maintain diagnostic accuracy. These results also indicate that 11 % animal prevalence of brucellosis did not exert a detrimental effect on 305-day milk yield and reproductive performance in high milk-yielding Holstein cows.     

Association between cattle herd Mycobacterium avium subsp. paratuberculosis (MAP) infection and infection of a hare population

Paratuberculosis has long been considered a disease of domestic and wild ruminants only. The known host range of Mycobacterium avium subsp. paratuberculosis (MAP) was recently extended to include non-ruminant wildlife species believed to be exposed to spillover of MAP from infected domestic cattle herds. The aim of the present study was to assess the association between cattle herd MAP infection pressure level and the infection level of a hare population in two dairy farms of southern Chile. Fifty hares from a herd A and 42 hares from herd B were captured and sampled for MAP culture. The results showed a statistically significant association between the cattle herds’ infection prevalence and the hare infection prevalence.     

Neospora caninum infection as a cause of reproductive failure in a sheep flock

Neospora caninum has been detected only sporadically in cases of ovine abortion, and it has therefore traditionally been considered as an unimportant parasite in small ruminants. This study was carried out with the aim of identifying the pathogen causing serious reproductive problems on a commercial sheep farm. Sera from all rams and ewes tested negative for antibodies against Border disease virus, Schmallenberg virus and Coxiella burnetii, and infections by these agents were therefore ruled out. Nevertheless, seropositivity to N. caninum and/or Toxoplasma gondii was detected, although the seroprevalence was higher in the case of N. caninum. The percentage of lambings and the number of lambs per dam were significantly lower in ewes that were seropositive to N. caninum while no effect on these parameters was detected in ewes that were seropositive to T. gondii. There was also no evidence of infection by T. gondii in the foetal/lamb tissues analyzed by PCR and/or immunohistopathological techniques. On the contrary, the DNA of N. caninum was detected in 13 out of 14 foetuses/lambs descendant from dams seropositive to this parasite. Characteristic lesions caused by N. caninum and/or its antigen were also detected. Genotyping of the N. caninum DNA revealed only two closely related microsatellite multilocus genotypes. The results clearly demonstrate that infection by N. caninum was the cause of the low reproductive performance of this sheep flock.     

Fragmentation patterns and systematic transitions of the forested landscape in the upper Amazon region,Ecuador 1990–2008

The analysis of the systematic transitions in the forested landscape and the study of the forest fragmentation patterns allow us to deepen our understanding of the changes in the vegetation ground cover. The importance of knowing the intricate patterns of the land usage of the upper basin of the Amazon region is widely recognized. This zone is one of the most diverse biological areas in the world, is home to large areas of mature tropical cloud forest and demonstrates high probabilities of stable climatic conditions in light of global warming. The research quan- tiffed systematic transitions through the ＂loss＂ and ＂gain＂ of the different categories of landscape during the eighteen-year study period of the Ecuadorian Amazon Region （EAR）, the forest fragmentation patterns were also analyzed based on a set of indicators. Therefore, with respect to the entirety of the landscape, the results registered for the ground coverage in forested areas during the first period （1990-2000）, show a decrease of 6.99% and an increase of 0.68%; and during the second period （2000-2008）, show a decrease of 3.99% and an increase of 2.14%. It demonstrated that forest and agricultural areas tended to replace or be replaced by herbaceous vegetation faster than expected fortuitously. Finally, the indices of fragmentation signaled intense changes during the 1990-2000 period with a reduction during the period 2000-2008. Per- centages registered in the Largest Patch Index （LPI） were between 79.58%; 52.39% and 49.99% respectively; while the Patch Density （PD） varied between 0.04; 0.06 and 0.07. This suggests the propensity of for- est cover to remain intact. The results of this investigation suggest a tendency towards stability in Ecuador＇s Amazon landscape. Within the framework for development and management of this area, the tendency is natural regeneration. This permits a consolidation of the conservation, reforestation, forestation and agricultural forestry plans, programs and systems for the protected areas in EAR.     

Distribution patterns of acorns after primary dispersion in a fragmented oak forest and their consequences on predators and dispersers

Forest fragmentation affects seed production, but little is known about how seeds are distributed in fragmented forests after primary dispersion and how this affects their interactions with other organisms. This study addressed these issues in a fragmented oak forest of Quercus laeta where four habitat types were recognized: interior and edge of patches, deforested matrix around patches, and an abandoned field. Areas and distances to the abandoned field were measured for 40 patches, and acorns were sampled in all habitats. Only samples from the interior and edge of patches had acorns, which were classified as aborted, viable, and parasitized by insects or fungi. Acorns in all these classes were regressed against patch area and distance to the abandoned field. Field experiments were also conducted to determine whether acorn removal rates varied across habitats. Aborted acorns within patches were not related to area or distance, but they decreased with increasing values of both variables at patch edges. Within patches, viable acorns were positively related to increases in area and distance, while the converse occurred for insect-damaged acorns; fungal-infested acorns were not related to patch variables in this habitat. Complex interactions between area and distance explained how acorns in these classes were distributed across patch edges. Acorn removal was higher within patches and their edges when compared to human-disturbed habitats; both mice and birds were identified as potential acorn dispersers. These results suggest that forest fragmentation affects acorn-mediated processes and that this may influence the recovery of patch connectivity in the landscape.