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.     

Anomalous Pregnancies during Late Embryonic/Early Foetal Period in High Producing Dairy Cows

This study analyses anomalous cases of gestation ending in pregnancy loss during the early foetal period and their effect on progesterone and plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations. Data derived from a large-scale ultrasound pregnancy diagnosis programme in high producing dairy cows. Over a 3-year period (2004–2007), a very low incidence (0.5%: 15 of 3094) of anomalous pregnancies was recorded. The results revealed that the following anomalies were detected on days 35–41 of gestation in cows carrying singletons with one single corpus luteum: embryo death in eight cows (0.3%); and embryo in the uterine horn contralateral to the corpus luteum in seven cows (0.2%). All these animals suffered pregnancy loss during the early foetal period. In cows carrying dead embryos, no signs of conceptus degeneration were observed on pregnancy diagnosis. Amnion size (approximately 25 mm diameter) and uterine horn fluid contents were estimated to be similar to those of the normal pregnant cows in this period. In the contralateral gestations, live embryos were observed in all ultrasound checks before pregnancy loss. Uterine fluid contents increased in the two cows in which gestation continued for more than a week. In the cases of embryo death but not in those of contralateral gestation, a drop in PAG-1 levels was noted prior to pregnancy loss. Two cows carrying dead embryos increased with time allantoic fluid contents. The PAG-1 values increased with time in one cow bearing a dead embryo (from 2.31 to 6.79 ng/ml) and in two of the contralateral gestations (from 1.66 to 2.33 ng/ml and from 0.39 to 6.79 ng/ml, respectively). Results of this study indicate that the foetal membranes continue to undergo some activity following embryo death, and that contralateral pregnancy may determine failure of the gestation process.     

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.     

Effects of Sperm Concentration and Straw Volume on Motion Characteristics and Plasma, Acrosomal, and Mitochondrial Membranes of Equine Cryopreserved Spermatozoa

The advantages of using cryopreserved semen in equine reproduction are well known. During cryopreservation, spermatozoa undergo many changes that lead to a decrease in fertility. There is no agreement on the ideal sperm dose and concentration to maximize fertility rates. Thus, the objectives of this experiment were to evaluate sperm motion by computer-assisted analysis (CASA), sperm membrane integrity and function with fluorescence probes of cryopreserved sperm at three concentrations: 100 (C100), 200 (C200) and 400 × 10⁶ sperm/mL (C400), and two straw volumes (0.50 and 0.25 mL). There was no interaction between sperm concentration and storage volume (P > .05). Sperm motion characteristics were influenced by concentration (C100 > C200 > C400; P < .05). Curvilinear velocity (VCL) in 0.25-mL straws had higher average values (P < .05). Membrane integrity and function were not changed by straw volume (P > .05). However, sperm concentration changed the percentage of cells with intact plasma membrane (C100 > C200 > C400; P < .05) and the percentage of cells with high mitochondrial membrane potential (C100 = C200; P > .05 and C400 < C100 and C200; P < .05). According to this experiment, the best freezing method was that involving 100 × 10⁶ sperm/mL, regardless of straw volume.     

Differentiation between canine cutaneous and visceral leishmaniasis by the detection of immunoglobulin G specific for Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi antigens using flow cytometry

Flow cytometry employing Leishmania (L.) chagasi (Lc) and L. (Viannia) braziliensis (Lb) antigen was used to establish the differential diagnosis between visceral (VL) and cutaneous leishmaniasis (CL) in dogs. Flow cytometry permitted the detection of Leishmania-specific immunoglobulin G in sera from 19 dogs: nine with CL and 10 with VL. A significant difference in the percentage of positive staining was observed in sera from dogs with CL between the homologous antigen (69% for Lb) and the heterologous antigen (42% for Lc). However, this difference was not significant in sera from dogs with VL (61% for Lb and 73% for Lc). No significant staining was observed in control sera (0.6% for Lb and 0.4% for Lc) consisting of samples from healthy dogs, or in the group with sporotrichosis (1.8% for Lb and 1.5% for Lc), a differential diagnosis of CL. The results suggest that flow cytometry might be useful for the differentiation between CL and VL in dogs, with practical applications in areas where the two infections overlap.     

Epidural injection of ketamine for caudal analgesia in the cow

The objective of the study was to determine the analgesic and sedative effects of epidural ketamine in the cow. Eight healthy cows weighing 350–450 kg were used. One of 3 doses of ketamine (0.5, 1 and 2.0 mg/kg) or a saline control were injected into the epidural space at the first intercoccygeal interspace in random sequence at one-week intervals. Ketamine was diluted in saline (0.9%) before the experiment, and the volume adjusted according to animal size. Analgesia was tested by applying a standard stimulus (needle insertion into the skin and deep muscle) and scored using a 3-point scale. A second voltage-based stimulus was also applied and the responses scored. Another scale was used for scoring the degree of sedation. The response and the degree of sedation were assessed before drug administration and at 2, 5, 10, 15 min, and every 15 min until 120 min after ketamine or saline administration. Ketamine produced dose-related analgesia of the tail, anus, perineum and vulva but not of the hindlimb area. The effect was dose-dependent in terms of intensity and duration. None of the doses produced analgesia when 70 or 80 volts were applied. Minimal side effects were observed. Epidural ketamine produces caudal analgesia in the cow. Further studies are required to determine whether this is sufficient for surgery.     

Aurantiochytrium sp. meal can replace fish oil in practical diets for the juvenile Pacific white shrimp

This study evaluated the use of microalgae (Aurantiochytrium sp.) meal as a substitute for fish oil in the diet of juvenile Pacific white shrimp (Litopenaeus vannamei) reared in a clear‐water system. Dietary treatments at five replacement levels (0%, 25%, 50%, 75%, 100%) were performed in triplicate. After 46 days, only a slight difference in shrimp final weight was observed among treatments (0.61 g). An increase in final weight was observed with replacement of up to 50% fish oil for microalgae meal, while the optimal percentage of replacement estimated was 44.7%. Feed conversion rate (FCR) of animals fed a diet of Aurantiochytrium sp.meal to replace up to 50% fish oil decreased, and the optimal percentage of replacement estimated was 49.3%. The fatty acids profile in shrimp muscle tissue demonstrated an increase in docosahexaenoic acid (DHA) from 10.03% to 14.28% with increased replacement of fish oil by microalgae meal in the diet. Therefore, the partial replacement of fish oil by microalgae meal resulted in improved shrimp growth and FCR, and total replacement of dietary fish oil had no negative effects on these parameters. In addition, inclusion of microalgae meal raises the level of DHA in shrimp muscle.     

Response of juvenile pacu (Piaractus mesopotamicus Holmberg, 1887) to balanced digestible protein

This 45‐day work aimed to determine the response of juvenile pacu (Piaractus mesopotamicus) to balanced digestible protein (BDP) and to use these responses to determine whether the optimum economic levels of BDP would differ depending on the form in which the fish is sold. Six isoenergetic diets containing 163, 201, 238, 272, 315 and 348 g/kg BDP (dry‐matter basis) were prepared through the serial dilutions of a high‐protein diet with the low‐protein diet. Fish (initial average body weight, 10.82 ± 0.14 g) were fed with respective experimental diets three times a day until apparent satiation. The optimum biological level of BDP was calculated as 326 g/kg (dry‐matter basis) by the quadratic regression model for maximum body weight gain of juvenile pacu. To maximize economic returns (US$/kg) for different end products, the optimum economic levels of BDP were calculated as 311, 317 and 319 g/kg (dry‐matter basis) by an economic model for whole‐body, eviscerated and sliced juvenile pacu respectively. This finding revealed that obtaining maximum biological performance of fish in the case of high feed costs or low prices of the end product in consumer market would substantially reduce the economic returns.     

Digestible protein requirements and muscle growth in juvenile tambaqui (Colossoma macropomum)

Several investigations have been carried out to improve the productivity of tambaqui, an economically important fish species in Brazil and other Latin American countries. This study determined the digestible protein (DP) requirements in juvenile tambaqui by assessing their productive performance and nutritional efficiency. It also evaluated the effects of different dietary DP levels on the morphology and cellularity of skeletal fast muscle fibres. The 1750 tambaqui tested (6.53 ± 0.43 g body weight, 7.58 ± 0.18 cm length) were randomly distributed into 35 tanks. Fish were fed one of the seven isocaloric diets, which contained 140, 170, 200, 230, 260, 290 or 320 g/kg DP. The DP requirement, calculated by segmented (broken line) regression of weight gain data, was 290 g/kg. An increase in diet DP to 290 g/kg significantly improved final weight, weight gain, feed intake, specific growth rate and crude protein gain, and changed fibre diameter in deep muscle. Muscle fibres were randomly distributed into a mosaic pattern, characterized by fibres with different diameters. Treatments with 290 and 320 g/kg DP increased the frequency of large‐diameter fibres (>50 µm), indicating hypertrophic growth of skeletal muscle during the juvenile phase, which occurred in conjunction with hyperplasia.