Butyrate and propionate improve the growth performance of Litopenaeus vannamei

Organic salts may improvement the animal performance, increasing the efficiency of nutrient utilization and modifying the intestinal microbiota. This study aimed to evaluate the effects of sodium butyrate and sodium propionate supplementation at different levels of dietary inclusions in the growth of Litopenaeus vannamei. In total, seven diets were evaluated: a control diet (without supplementation) and three diets from each sodium salt, propionate and butyrate, in concentrations of 0.5%, 1% and 2%. We used 21 tanks of 6000 L stocked with 150 shrimps (2.53 ± 0.03 g). The shrimps fed diets supplemented with propionate and butyrate, in all concentrations, increased their final weight. The feed efficiency, nitrogen retention, protein efficiency rate, survival and yield of shrimps fed the diet containing 2% butyrate were higher in comparison with control treatment. The shrimps supplemented with butyrate also showed lower counts of Vibrio sp. in the intestine. The shrimps fed the diet supplemented with butyrate and propionate also showed higher values of serum agglutination titre. Thus, it is possible to conclude that dietary supplementation with propionate and butyrate in different dietary concentrations modify the intestinal microbiota and improves the growth of L. vannamei.     

Na+/K+-ATPase Activation by cAMP in the Midgut of Lutzomyia longipalpis (Lutz & Neiva, 1912; Diptera: Psychodidae)

Lutzomyia longipalpis (Lutz & Neiva, 1912) females have been intensively studied regarding the regulation of midgut pH. The mechanisms involved in pH regulation are complex, and some aspects remain to be clarified. Here, we investigated the role of the Na⁺/K⁺-ATPase pump as an electrochemical potential generator and its modulation by the second messenger cAMP in the midgut of female L. longipalpis. Our results suggest that not only may Na⁺/K⁺-ATPase be the main generator of an electrochemical potential across membranes in the midgut of female L. longipalpis, but also its activity is positively regulated by cAMP. cAMP-mediated Na⁺/K⁺-ATPase pump activity might be necessary to maintain the transport of the nutrients produced during blood digestion.     

Ultrasound‐assisted periconal ocular blockade in rabbits

ObjectiveThis study aimed to evaluate the benefit and specifically the feasibility of using ultrasound in ophthalmologic periconal block, and the occurrence of complications.Study designProspective experimental study.AnimalsTen healthy New Zealand White rabbits (6–8 months of age), weighing 2.0–3.5 kg.MethodsRabbits were anesthetized by intramuscular injection of acepromazine (1 mg kg⁻¹), ketamine (30 mg kg⁻¹) and xylazine (3 mg kg⁻¹). Ultrasound-assisted periconal block with lidocaine was performed on 18 eyes. Intraocular pressure was measured by applanation tonometry whereas corneal sensitivity was assessed using an esthesiometer, before and after each periconal anesthesia.ResultsIn all 18 eyes, it was possible to adequately visualize the needle shaft within the periconal space, as well as muscular cone, optic nerve and local anesthetic solution spread. Lidocaine 2% without epinephrine (0.79 ± 0.19 mL) was injected into the periconal space. There was no statistical difference between the intraocular pressure (mean ± SD) measured before (10.9 ± 2.9 mmHg) and after (11.9 ± 3.8 mmHg) the periconal anesthesia (p = 0.38). The effectiveness of the ultrasound-assisted technique was shown according to the values for corneal sensitivity, assessed before and after periconal anesthesia (p < 0.0001). Complications were not observed in this study.ConclusionsEye ultrasonography allowed visualization of all anatomic structures necessary to perform a periconal block, as well as the needle insertion and anesthetic spread in real time. Further studies are required to prove the real potential of ultrasound for reducing the incidence of complications associated with ophthalmic blocks, especially when anatomic disorders of the eye could potentially increase the risk.Clinical relevanceUltrasonography is a painless, noninvasive tool that may improve safety of ophthalmic regional blocks, potentially by reducing the prevalence of globe perforation or penetration of the optic nerve associated with the needle-based techniques.     

Inferred longevity of Amazonian rainforest trees based on a long-term demographic study

We used data from a long-term (14–18 years) demographic study to infer the maximum longevity for populations of 93 relatively abundant tree species in central Amazonia. We also assessed the influence of several life-history features (wood density, growth form, mortality rate, recruitment rate, stem diameter, growth increment, population density) on tree longevity. Data on 3159 individual trees were collected in 24 permanent, 1 ha plots in undisturbed forest arrayed across a large (ca. 1000 km²) study area. For each species, three estimates of longevity were generated (by dividing the stem diameter of the largest tree by the median, upper quartile, and upper decile of observed diameter-growth rates), and the mean of these three values was used as a longevity estimate. Longevity values ranged from 48 years in the pioneer Pourouma bicolor (Cecropiaceae) to 981 years for the canopy tree Pouteria manaosensis (Sapotaceae), with an overall mean of 336±196 years. These growth-based estimates of maximum tree age were concordant with those derived from analyses of mean mortality rates. Tree longevity was positively correlated with wood density, maximum stem diameter, and population density, and negatively correlated with annual mortality, recruitment, and growth rates. On average, pioneer species had much lower longevity than did non-pioneers, whereas among old-growth trees, emergent species had greater longevity than did canopy species. Our results are consistent with radiocarbon-based studies that suggest that Amazonian trees can occasionally exceed 1000 years of age.     

Kinetics of energy source utilization in Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) embryonic development

The present work evaluates the kinetics of utilization of the main potential energy sources throughout the embryonic developmental stages of Boophilus microplus. The embryonic development of this arthropod is completed in 21 days. Cellularization of the blastoderm occurs on the 6th day and is rapidly followed by germ band extension and segmentation, whose first signs are visible on the 7th day. Cellularization is typically a maternal-driven process, carried out by molecular determinants deposited in the oocyte during oogenesis. On the other hand, segmentation is of zygotic nature, being the consequence of the synthesis of various components by the growing embryo. The enhancement in total B. microplus RNA was observed after cellularization, corroborating the replacement of maternal-driven processes by embryonic zygotic expression. An abrupt increase in oxygen consumption was observed from cellularization until the 8th day of development. The reduction in dry weight at the same period and the susceptibility of oxygen consumption to KCN suggest that the respiration process is activated during early embryonic development. A marked decrease in total lipid content occurred between the 5th and 7th days of development, suggesting this is the main energy source for cellularization. A major reduction in carbohydrate content occurred later, between the 7th and 9th days, and it could be assigned to the morphological segmentation of the embryo. Although the total amount of proteins remains unchanged from oviposition to hatching, a 15% reduction in vitellin (VT) content was observed before cellularization, up to the 4th day after egglaying. This observation was correlated to the synthesis of new proteins needed to support early embryo development. Additional 20% of VT was consumed thereafter, mainly at the end of embryogenesis, and in this case VT is probably used as energy source to the older embryo. Altogether, these data indicate different energy sources for maternal and zygotic driven processes.     

Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs

Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.     

First report and molecular characterization of Trypanosoma vivax in cattle from state of Pernambuco, Brazil

This paper describes an outbreak of Trypanosoma vivax for the first time in the state of Pernambuco, Brazil, affecting dairy cattle in the municipality of Itambé in the northern coastal zone of the state. Clinical signs compatible with infection by blood protozoa and epidemic miscarriages were observed. The diagnosis of T. vivax was confirmed through biometric microscopy and molecular analysis with PCR and DNA sequencing. The T. vivax isolate detected in the present study proved to be genetically very close to other Brazilian isolates of the protozoan despite being geographically distant.     

Supplementation of citric acid in plant protein‐based diets for juvenile tambaqui,Colossoma macropomum

We evaluated citric acid supplementation (0, 10, 20, and 30 g/kg) in plant protein‐based diets for juvenile tambaqui, Colossoma macropomum. In Experiment 1, fish (n = 160; 27.56 ± 2.73 g) were distributed in 310 L tanks (n = 16) for 80 days. Zootechnical performance, hematological parameters, and mineral composition (muscle and bones) did not differ significantly (p > .05). The inclusion of citric acid‐reduced pH of diets (p = .001) and stomach (p = .035). In Experiment 2, fish (n = 60; 48.00 ± 1.98 g) were distributed in fecal collection aquaria (200 L) for 21 days. The following were observed: a linear effect for the crude protein (p = .0004) and crude fat (p = .0037) digestibility coefficients; a quadratic effect for crude energy (p = .0037) and dry matter (p = .0004); an optimal supplementation level of 18.5 g/kg. Calcium (p = .0060) and magnesium (p = .0222) showed a linear effect, while phosphorus (p = .0001) had a quadratic effect with the 20 g/kg optimum supplementation level. To conclude, the supplementation of this organic acid in diets does not negatively affect fish nutritional status, health or welfare, and increases nutrient and mineral availability.     

Detection and Characterization of Hepatitis E Virus in Domestic Pigs of Different Ages in Portugal

This study represents the primary hepatitis E virus (HEV) surveillance in domestic pigs in Portugal, five pig farms were investigated in 5 different Portuguese regions, ten faecal samples were collected at four different stages of the production. All faecal samples were tested for hepatitis E virus by real-time RT-PCR. At least one sample from each farms of all age groups tested positive for HEV. The prevalence in the pig herds varied from 10% to 30% and the mean prevalence was 32% in weaners, 20% in growers, 32% in fatteners and 4% in adult dry sows. Phylogenetic analysis of the detected HEV sequences indicated that the circulating virus strains belong under the genotype 3.     

Immunogenicity in dogs of three recombinant antigens (TSA, LeIF and LmSTI1) potential vaccine candidates for canine visceral leishmaniasis

Control of canine visceral leishmaniasis (VL) remains a difficult and serious problem mostly because there is no reliable and effective vaccine available to prevent this disease. A mixture of three recombinant leishmanial antigens (TSA, LeIF and LmSTI1) encoded by three genes highly conserved in the Leishmania genus have been shown to induce excellent protection against infection in both murine and simian models of cutaneous leishmaniasis. A human clinical trial with these antigens is currently underway. Because of the high degree of conservation, these antigens might be useful vaccine candidates for VL as well. In the present study, using the dog model of the visceral disease, we evaluated the immunogenicity of these three antigens formulated with two different adjuvants, MPL-SE and AdjuPrime. The results were compared with a whole parasite vaccine formulated with BCG as the adjuvant. In order to investigate if sensitization with the recombinant antigens would result in recognition of the corresponding native parasite antigens upon infection, the animals were exposed for four weeks after the termination of the immunization protocol with the recombinant antigens to a low number of L. chagasi promastigotes, an etiological agent of VL. Immune response was evaluated by quantitative ELISA in the animal sera before and after exposure to the viable parasites. Both antigen specific IgG1 and IgG2 antibody levels were measured. Immunization of dogs with the recombinant antigens formulated in either MPL-SE or AdjuPrime resulted in high antibody levels particularly to LmSTI1. In addition, this immunization although to low levels, resulted in the development of antibody response to the whole parasite lysate. Importantly, experimental exposure with low numbers of culture forms of L. chagasi promastigotes caused a clear boost in the immune response to both the recombinant antigens and the corresponding native molecules. The boost response was predominantly of the IgG2 isotype in animals primed with the recombinant antigens plus MPL-SE. In contrast, animals primed with the recombinant antigens formulated in AdjuPrime as well as animals vaccinated with crude antigen preparation responded with mixed IgG1/IgG2 isotypes. These results point to the possible use of this antigen cocktail formulated with the adjuvant MPL-SE in efficacy field trials against canine VL.