Replacement of fishmeal by vegetable meal mix in the diets of Litopenaeus vannamei reared in low‐salinity biofloc system: effect on digestive enzymatic activity

Biofloc (consortium of diverse microorganisms associated to suspending substrates) was developed from waste of shrimp Litopenaeus vannamei postlarvae culture under low salinity (5 g L^?1) to provide an additional nutritious biomass and reduce fishmeal inclusion in feeds in a 28‐day indoor shrimp nursery trial conducted in 15 experimental containers (250 L stocked at 600 org m^?3). Four experimental diets (isoproteic and isocaloric) containing different percentage of fishmeal: 0%, 10%, 20% and 30% substituted by vegetable meal mix (corn, sorghum and wheat) were formulated and elaborated. A control treatment consisted of a commercial feed. The main water quality parameters were monitored, and no significant differences were found among treatments. The growth and survival were similar among treatments. In general, digestive enzymatic activities showed differences being greater in the biofloc system compared with clear water. It was concluded that low‐salinity shrimp nursery could be successfully developed with minimum inclusion of fishmeal in feeds, without significant effect on production response. The adjustment of C : N ratio allowed the increase of microbial biomass in the bioflocs, which contributed to maintain good water quality, provide live food and enhance digestive enzymatic activity of cultured organisms.     

Receiver operating characteristic-based assessment of a serological test used to detect Johne’s disease in Israeli dairy herds

The overall accuracy of an enzyme-linked immunosorbent assay (ELISA) used to detect Johne's disease at herd level was explored in relation to an imperfect test (fecal culture) in 57 Israeli dairy herds. Receiver-operating characteristic (ROC) analysis indicated an area under the curve (AUC) that corresponded to a test accuracy of 82.0% (69.5% to 90.9%; 95% confidence), with optimized herd sensitivity and herd specificity of 70.4% and 83.3%, respectively; and predictive values of 79.2 (+) and 75.8% (-). The optimal ELISA cutoff was 3.16% (> 3.16% seropositive cows in a herd), which was associated with likelihood ratios (LR) of 4.22 (+LR) and 0.36 (-LR), and post-test probabilities of 0.79 (+) and 0.17 (-). For herds with < or = 200 cows (n = 19 herds), the 95% confidence interval (CI) for the AUC was 0.62-0.97 and the optimal cutoff was 3.33% (HSe = 87.5, HSp = 81.8); for herds with > 200 but < or = 270 cows (n = 19 herds), the 95% AUC CI was 0.62-0.97 and the optimal cutoff was 1.13% (HSe = 90.0, HSp = 77.78); and for herds with > 270 cows (n = 19 herds), the 95% AUC CI was 0.69-0.99 and the optimal cutoff was 0.7% (HSe = 100.0, HSp = 70.0). The AUC was not influenced by across-herd prevalence [R2 (adjusted) = 0.0, P > 0.05]. Findings may be applied to facilitate targeted sampling of herds similar to those evaluated. For instance, a test cutoff of 0.76% could be considered for "ruling disease in," while a cutoff of 3.7% could be used for "ruling disease out." Caveats that may influence this analysis are discussed.     

Development of a Model to Simulate Nitrogen Dynamics in an Integrated Shrimp–Macroalgae Culture System with Zero Water Exchange

The effluents of traditional shrimp monoculture cause pollution and promote eutrophication and hypernutrification of the receiving coastal ecosystems. Integrated aquaculture and a recirculating aquaculture system (RAS) have been proposed as an alternative to address these problems. In this study, we developed a dynamic model to simulate the concentration of total ammonia nitrogen (TAN), nitrite, and nitrate in an integrated culture of whiteleg shrimp, Litopenaeus vannamei, and seaweed, Gracilaria vermiculophylla, in a recirculating and zero water exchange system, and the effect of nitrifying and heterotrophic bacteria was also included. The experiments demonstrated that a dynamic model can explain the concentrations of dissolved inorganic nitrogen and variations in these concentrations over time in the integrated culture. The results also suggest that nitrifying and heterotrophic bacteria play an important role in the transformation of dissolved nitrogenous compounds; therefore, these bacteria should be considered within the dynamics of nitrogen in integrated systems with low water exchange.     

The NS segment of H5N1 avian influenza viruses (AIV) enhances the virulence of an H7N1 AIV in chickens

Some outbreaks involving highly pathogenic avian influenza viruses (HPAIV) of subtypes H5 and H7 were caused by avian-to-human transmissions. In nature, different influenza A viruses can reassort leading to new viruses with new characteristics. We decided to investigate the impact that the NS-segment of H5 HPAIV would have on viral pathogenicity of a classical avian H7 HPAIV in poultry, a natural host. We focussed this study based on our previous work that demonstrated that single reassortment of the NS-segment from an H5 HPAIV into an H7 HPAIV changes the ability of the virus to replicate in mammalian hosts. Our present data show that two different H7-viruses containing an NS-segment from H5–types (FPV NS GD or FPV NS VN) show an overall highly pathogenic phenotype compared with the wild type H7–virus (FPV), as characterized by higher viral shedding and earlier manifestation of clinical signs. Correlating with the latter, higher amounts of IFN-β mRNA were detected in the blood of NS-reassortant infected birds, 48 h post-infection (pi). Although lymphopenia was detected in chickens from all AIV-infected groups, also 48 h pi those animals challenged with NS-reassortant viruses showed an increase of peripheral monocyte/macrophage-like cells expressing high levels of IL-1β, as determined by flow cytometry. Taken together, these findings highlight the importance of the NS-segment in viral pathogenicity which is directly involved in triggering antiviral and pro-inflammatory cytokines found during HPAIV pathogenesis in chickens.     

The role of spatial mixing in the spread of foot-and-mouth disease

A model of epidemic dispersal (based on the assumption that susceptible cattle were homogeneously mixed over space, or non-spatial model) was compared to a partially spatially explicit and discrete model (the spatial model), which was composed of differential equations and used geo-coded data (Euclidean distances between county centroids). While the spatial model accounted for intra- and inter-county epidemic spread, the non-spatial model did not assess regional differences. A geo-coded dataset that resembled conditions favouring homogeneous mixing assumptions (based on the 2001 Uruguayan foot-and-mouth disease epidemic), was used for testing. Significant differences between models were observed in the average transmission rate between farms, both before and after a control policy (animal movement ban) was imposed. They also differed in terms of daily number of infected farms: the non-spatial model revealed a single epidemic peak (at, approximately, 25 epidemic days); while the spatial model revealed two epidemic peaks (at, approximately, 12 and 28 days, respectively). While the spatial model fitted well with the observed cumulative number of infected farms, the non-spatial model did not (P<0.01). In addition, the spatial model: (a) indicated an early intra-county reproductive number R of approximately 87 (falling to <1 within 25 days), and an inter-county R<1; (b) predicted that, if animal movement restrictions had begun 3 days before/after the estimated initiation of such policy, cases would have decreased/increased by 23 or 26%, respectively. Spatial factors (such as inter-farm distance and coverage of vaccination campaigns, absent in non-spatial models) may explain why partially explicit spatial models describe epidemic spread more accurately than non-spatial models even at early epidemic phases. Integration of geo-coded data into mathematical models is recommended.     

Necrotizing Enterocolitis in Horses: A Retrospective Study

The clinical and clinicopathologic characteristics of fatal necrotizing enterocolitis were examined in 16 horses (age 4 months to 12 years). At initial presentation, 8 of 16 horses were pyrexic (median temperature, 38.4°C; range, 33.8 to 40.6°C); all 16 were tachycardic (median heart rate, 93 bpm, range, 66 to 138 bpm); 13 of 16 were tachypneic (median heart rate, 36 bpm, range, 16 to 80 bpm), dehydrated, and had discolored mucous membranes. All horses that were pyrexic were also tachycardic and tachypneic. PCV was high (>45%) in 14 horses. Six horses were leukopenic (<5,000 cells/μL); 12 were neutropenic (<2,300 cells/μL), and 14 had >100 band neutrophils/μL. Twelve horses were acidemic (pH 7.37; range, 6.88 to 7.33) and the venous bicarbonate concentration was low (<23 mEq/L) in 14 horses. Median anion gap in 16 horses was 31.5 mEq/L (>15 mEq/L in 15 horses). Eleven of 16 horses were hyponatremic (<137 mEq/L), 1 horse was hypernatremic (> 143 mEq/L), 3 were hypoka-(emic (<3.2 mEq/L), 6 were hyperkalemic (>4.5 mEq/L), and 14 were hypochloremic (<98 mEq/L). Serum creatinine concentrations were high (>1.4 mg/dL) in 15 horses. Abdominal fluid was examined in 12 horses: 4 had total protein concentrations 2.5 g/dL and 6 had nucleated cell counts >5,000/μL and <10,000/μL; none had >10,000/μL. Eight of 12 samples revealed a nondegenerate neutrophilia (>50%). Abdominal fluid collected from 4 horses immediately before death was normal in 2 horses and indicative of suppurative inflammation in 2. All 8 horses tested had low or nonexistent serum immunofluorescent antibody titers to Ehrlichia risticii. Four of 16 horses had Salmonella spp isolated from feces or tissues. All 16 horses either died (5 of 16; 31%) or were euthanized because of a grave prognosis. Median time to death was 45.5 hours (range, 7 to 113 hours) from the time of admission. Death was preceded by severe abdominal pain in 14 horses. Fatal necrotizing enterocolitis of horses is characterized by a brief course, profound dehydration, electrolyte derangements, acid-base abnormalities, and terminally, severe abdominal pain. Abdominal fluid analysis was frequently not indicative of the severity of disease. J Vet Intern Med 1996;10:265–270. Copyright©1996 by the American College of Veterinary Internal Medicine.     

Response of lactating dairy cows fed different supplemental zinc sources with and without evaporative cooling to intramammary lipopolysaccharide infusion: metabolite and mineral profiles in blood and milk

The objective of this study was to determine the effect of evaporative cooling and dietary supplemental Zn source on blood metabolites, insulin and mineral concentrations, and milk mineral concentrations following intramammary lipopolysaccharide (LPS) infusion. Seventy-two multiparous Holstein cows were assigned to one of four treatments with a 2 × 2 factorial arrangement. Treatments included two environments: with or without evaporative cooling using fans and misters over the freestall and feedbunk, and two dietary sources of supplemental Zn: 75 mg/kg of dry matter (DM) supplied by Zn hydroxychloride (inorganic Zn; IOZ) or Zn hydroxychloride (35 mg of Zn/kg of DM) + Zn–Met complex (ZMC; 40 mg of Zn/kg of DM). A subset of cows (n = 16; 263 ± 63 d in milk) was infused with 10 μg of LPS or a saline control in the left or right rear quarters on day 34 of the environmental treatment. Individual milk samples collected from LPS-infused quarters at −4, 0, 6, 12, 24, 48, 72, 96, and 144 h relative to infusion were analyzed for minerals. Blood samples were collected at the same time with an additional sample collected at 3 h post-infusion to analyze glucose, nonesterified fatty acids (NEFA), insulin, and minerals. Cooling by time interactions (P ≤ 0.07) were observed for plasma glucose, NEFA, and serum insulin. Compared with cooled cows, non-cooled cows had lower concentrations of plasma glucose except at 3 h following intramammary LPS infusion, greater serum insulin at 3 and 12 h, and lower plasma NEFA at 24 and 48 h after infusion. Relative to cooled cows, non-cooled cows tended (P = 0.07) to have lower serum K concentration and had lower (P < 0.01) serum Zn 6 h following infusion (cooling by time interaction: P < 0.01). Relative to ZMC cows, IOZ cows had greater (P ≤ 0.09) concentrations of plasma Se, skim milk Na and Se, and skim milk Na to K ratio. Regardless of treatment, intramammary LPS infusion reduced (P < 0.01) serum or plasma concentrations of Ca, Mg, Zn, Fe, and Se, but increased (P < 0.01) their concentration in skim milk. In conclusion, deprivation of cooling resulted in more rapid and prolonged insulin release and influenced the systemic and mammary mineral metabolism during mammary inflammation induced by LPS of lactating dairy cows. Dietary supplementation of Zn–Met complex reduced blood and milk Se concentrations compared with cows fed Zn from an inorganic source.     

Determination of 67Zn distribution in navy bean (Phaseolus vulgaris L.) after foliar application of 67Zn-lignosulfonates using isotope pattern deconvolution

The improvement of Zn fertilizers requires new techniques to evaluate their efficacy. In this paper, the (67)Zn stable isotope was used as tracer of several Zn-lignosulfonate complexes to study the foliar-applied Zn uptake and distribution behavior in the plant, compared with ZnEDTA. Navy bean plants ( Phaseolus vulgaris L.) were grown hydroponically in a Zn-free nutrient solution, and six modified lignosulfonates and EDTA complexed with (67)Zn were used in foliar application in the young leaves as Zn sources. Zinc isotopes in roots, stems, and sprayed and unsprayed leaves were determined by ICP-MS, and signal interferences caused by the compounds of the digested vegetal samples were corrected. The mathematical procedure of isotope pattern deconvolution allowed the minimization of the uncertainty in the measured molar fractions of Zn from fertilizer or from natural sources. Significant differences in Zn use and distribution were observed among the fertilizers when the calculated concentrations of Zn from the fertilizer were compared, whereas they were unnoticeable attending to the total Zn in plant tissues, usually determined at the conventional studies. By foliar spray, higher Zn uptake and mobilization to leaves and stems were achieved with (67)ZnEDTA than with (67)Zn-LS complexes. The ultrafiltered LS and phenolated LS showed slightly better ability to provide Zn to the bean plants than the other LS. The foliar-applied Zn use and distribution in the plant were related with the stability of the Zn-lignosulfonates complexes. Those presenting the lower stability versus pH, but the highest complexing capacity, were slightly more suitable to supply foliar-applied Zn to navy beans.     

Characterization of a benzyl alcohol dehydrogenase from Lactobacillus plantarum WCFS1

Aroma is an important sensory parameter of food products. Lactic acid bacteria have enzymatic activities that could be important in the modification of food aroma. The complete genome sequence from Lactobacillus plantarum WCFS1 shows a gene (lp_3054) putatively encoding a protein with benzyl alcohol dehydrogenase activity. To confirm its enzymatic activity lp_3054 from this strain has been overexpressed and purified. Protein alignment indicated that lp_3054 is a member of the family of NAD(P)-dependent long-chain zinc-dependent alcohol dehydrogenases. In lp_3054 all of the residues involved in zinc and cofactor binding are conserved. It is also conserved the residue that determines the specificity of the dehydrogenase toward NAD (+) rather than NADP (+) and, therefore, L. plantarum benzyl alcohol dehydrogenase is less active in the presence of NADP (+) than in the presence of NAD (+). The purified enzyme exhibits optimal activity at pH 5.0 and 30 degrees C. The kinetic parameters K m and V max on benzyl alcohol as a substrate were, respectively, 0.23 mM and 204 mumol h (-1) mg (-1). Besides its activity toward benzyl alcohol, it showed activity against nerol, geraniol, phenethyl alcohol, cinnamyl alcohol, and coniferyl alcohol, all of which are volatile compounds involved in determining food aroma. The biochemical demonstration of a functional benzyl alcohol dehydrogenase activity in this lactic acid bacteria species should be considered when the influence of bacterial metabolism in the aroma of food products is determined.