Effect of a Lactobacillus animalis strain on composition and metabolism of the intestinal microflora in adult dogs

Probiotics are microorganisms that are added to food to exert beneficial effects on the host. Aim of the present study was the in vitro and in vivo evaluation of the effect of Lactobacillus animalis LA4 (isolated from the faeces of a healthy adult dog) on composition and metabolism of dog intestinal microflora. When added to dog faecal cultures, LA4 reduced enterococci and increased lactobacilli counts throughout the study, whereas C. perfringens counts were significantly reduced at 24 h. After 8 h of incubation, LA4 reduced ammonia and increased lactic acid concentrations. For the in vivo study, nine adult dogs received the freeze-dried preparation of L. animalis LA4 for 10 days. On day 11, faecal lactobacilli were higher than at trial start (6.99 log CFU/g versus 3.35 log CFU/g of faeces) and faecal enterococci showed a trend towards a numerical reduction (P = 0.08). L. animalis LA4 was recovered in all faecal samples collected on day 11 and in four samples at day 15. The present results show that LA4 was able to survive gastrointestinal passage and transitorily colonize the dog intestine where, based on the in vitro results, it could positively influence composition and metabolism of the intestinal microflora. These results suggest that L. animalis LA4 can be considered as a potential probiotic for dogs.     

Neutral detergent-soluble fiber improves gut barrier function in twenty-five-day-old weaned rabbits

The effect of neutral detergent-soluble fiber level on gut barrier function and intestinal microbiota was examined in weaned rabbits. A control diet (AH) containing 103 g of neutral detergent-soluble fiber/ kg of DM included alfalfa hay as main source of fiber. Another diet (B-AP) was formulated by replacing half of the alfalfa hay with a mixture of beet and apple pulp resulting in 131 g of soluble fiber/kg of DM. A third diet (OH) was obtained by substituting half of the alfalfa hay with a mix of oat hulls and a soybean protein concentrate and contained 79 g of soluble fiber/kg of DM. Rabbits weaned at 25 d and slaughtered at 35 d were used to determine ileal digestibility, jejunal morphology, sucrase activity, lamina propria lymphocytes, and intestinal microbiota. Suckling 35-d-old rabbits were used to assess mucosa morphology. Mortality (from weaning to 63 d of age) was also determined. Villous height of the jejunal mucosa increased with soluble fiber (P = 0.001). Rabbits fed with the greatest level of soluble fiber (BA-P diet) showed the highest villous height/ crypt depth ratio (8.14; P = 0.001), sucrase specific activity (8,671 mumol of glucose/g of protein; P = 0.019), and the greatest ileal starch digestibility (96.8%; P = 0.002). The opposite effects were observed in rabbits fed decreased levels of soluble fiber (AH and OH diets; 4.70, 5,848 mumol of glucose/g of protein, as average, respectively). The lowest ileal starch digestibility was detected for animals fed OH diet (93.2%). Suckling rabbits of the same age showed a lower villous height/crypt depth ratio (6.70) compared with the B-AP diet group, but this ratio was higher than the AH or OH diet groups. Lower levels of soluble fiber tended (P = 0.074) to increase the cellular immune response (CD8+ lymphocytes). Diet affected IL-2 production (CD25+, P = 0.029; CD5+CD25+, P = 0.057), with no clear relationship between soluble fiber and IL-2. The intestinal microbiota biodiversity was not affected by diets (P >/= 0.38). Rabbits fed the B-AP and AH diets had a reduced cecal frequency of detection compatible with Campylobacter spp. (20.3 vs. 37.8, P = 0.074), and Clostridium perfringens (4.3 vs. 17.6%, P = 0.047), compared with the OH diet group. Moreover, the mortality rates decreased from 14.4 (OH diet) to 5.1% (B-AP diet) with the increased presence of soluble fiber in the diet. In conclusion, increased levels of dietary soluble fiber improve mucosal integrity and functionality.     

Isolation and development of haematopoietic progenitor cells from peripheral blood of adult and newborn pigs

Pluripotent stem cells (PSCs), already described in human beings, are fibroblast-like cells that exhibit a CD34 marker specific for haematopoietic stem cells. In this work we have demonstrated the presence of PSCs in the peripheral blood of pigs, a species frequently used in transplantation studies as an animal model for human diseases. Differentiation into haematopoietic colonies (granulomacrophagic colonies, erythroid colonies and mixed colonies) has been carried out with the peripheral blood of adult and newborn pigs, using solely human commercial media. Peripheral blood mononuclear cells (PBMNCs) were cultured in semisolid methylcellulose based media enriched with recombinant human cytokines, achieving granulomacrophagic-colony forming unit (GM-CFU) and mixed-colony forming unit (Mix-CFU) growth with erythroblastic lineage proliferation in the presence of erythropoietin (Epo). In all the samples CFU growth was associated with the presence of recombinant human cytokine. No evidence of proliferation in control plates without cytokines was found. From liquid medium culture, a population of macrophages and CD34+ fibroblast like cells were retrieved 21 days after sowing. These findings allow us to think about the direct application of this simple and standardised method in several work fields such as the study of pharmacological effects of many drugs over the haematopoietic line and in the study of new strategies in cellular therapy for some human diseases.     

Hematologic and biochemical changes during canine agility competitions

BACKGROUND: Normal response to different types of exercise needs to be established before performance, training level, or exercise intolerance can be assessed. OBJECTIVES: The aim of this research was to describe the hematologic and biochemical changes that dogs undergo during an agility competition. METHODS: Fifteen dogs were subjected to an agility test that covered a distance of 360-400 m with 40 obstacles. Basal venous blood samples were drawn 1 week before the competition, immediately after exercise, and at 5, 15, and 30 minutes of recuperation. A CBC, plasma biochemical profile, and lactate concentration were determined by standard methods using automated analyzers. Serum cortisol concentration was measured by competitive immune-enzyme analysis; insulin was measured by sandwich ELISA. RESULTS: Hematologic response to the agility test consisted of significant increases in RBC count, hemoglobin concentration, and HCT, with no change in total protein concentration. Serum triglyceride concentrations were increased postexercise, with highest values observed at 30 minutes of recuperation. Postexercise lactate concentrations exceeded the anaerobic threshold of 4 mmol/L. Serum lactate dehydrogenase activity was significantly higher after 30 minutes of recuperation. Significant differences were not observed in other analytes. CONCLUSIONS: Our results indicate that agility competitions induce mild to moderate changes in hematologic and biochemical results consistent with splenic contraction, increased lipolysis, and utilization of anaerobic pathways involved in energy resynthesis in muscle.     

Impact of feeding 2-hydroxy-4-(methylthio)butanoic acid and DL-methionine supplemented maize-soybean-rapeseed meal diets on growth performance and carcase quality of broilers

1. This experiment was conducted to compare the effectiveness of DL-2-hydroxy-4-(methylthio)butanoic acid (HMTBA) and DL-methionine (DLM) on growth performance and carcase quality of broilers in a 2 x 3 x 2 factorial experimental design including two methionine (Met) sources, three equimolar levels of Met supplementation and two sexes. 2. No difference was observed between the two Met sources in growth performance and muscle deposition from 10 to 49 d. With increasing Met levels, average daily gain was increased in the starter, grower and overall phases, average daily feed intake was increased in the starter phase, and feed:gain ratio was decreased in the grower and overall phases. Met supplementation increased breast muscle content and decreased abdominal fat content. Broilers given HMTBA had breast and thigh muscle coloration that was characterised as superior to those fed DLM. 3. These results indicated that Met supplementation improved growth performance and carcase quality irrespective of the Met sources. Compared to DLM, HMTBA has the same molar bioefficacy for improving the growth performance and carcase quality; however, HMTBA-fed birds had superior meat colour to DLM-fed birds.     

Effects of immunization of Pelibuey lambs with Oestrus ovis digestive tract protein extracts on larval establishment and development

Larval midgut proteins of hematophagous parasites contain strong antigens that can be used for host immunization. This concept has been applied for immunization of Pelibuey sheep against Oestrus ovis L. (Diptera: Oestridae). The aim of this study was to examine the effect of immunization on larval establishment (LE) and development. Immunized lambs (I, n = 6) received two injections of crude gut membrane protein extracts (GMPE) from third instar larvae with Freund's incomplete adjuvant (FIA) on days 0 (Day of first immunization) and 21 (0.4 and 0.45 mg GMPE/lamb, respectively). The control group (C, n = 5) received physiological saline with FIA. Lambs were challenged with first instars on Day 29 (20 larvae) and Day 43 (25 larvae). Blood samples were collected biweekly and IgG titers were analyzed by ELISA. All lambs were slaughtered on Day 90 and number of larvae recovered, larval stage and larval weight were recorded at necropsy. No significant effect of immunization on LE (C = 28.9%; I = 31.0% P > 0.05) was observed. Antibody titers were higher in the immunized group on Day 28 (P < 0.05), but subsequently similar in both groups. Larval physiological age and weight were also significantly (P < 0.05) affected by immunization. Immunization of Pelibuey lambs with GMPE did not affect LE but did delay O. ovis larval development.     

Infection by Leishmania infantum in cats: epidemiological study in Spain

More than 40 cases of feline leishmaniasis have been reported in the scientific literature. The influence of some immunodepressive conditions of viral origin, such as leukemia and feline immunodeficiency, are still unknown. The purpose of this study was to assess the prevalence of Leishmania infection in cats and possible relations with these viral infections. Markers of Leishmania infection were searched in 183 cats from Southern Spain by IFAT, PCR, Giemsa stain and culture, with a follow-up of positive cats. Seropositivity was 60.0% (Ab titer > or =10) and 28.3% of animals presented Ab titers > or =40. Around 25.7% of the cats studied were parasitemic and some of them remained positive for months. Combining both data, 70.6% of the feline population was, or could be, infected. We observed a negative association between seropositivity to Leishmania and infection by FeLV. Hence, production of antibodies against the parasite appears to be compromised in cats with leukemia, which have a prevalence of 36% in our study. In contrast, we found no association with feline immunodeficiency. The results makes us doubt the value of conventional serological methods to detect active Leishmania infection in cats.     

Assessment of climatic and orographic conditions on the infection by Calicophoron daubneyi and Dicrocoelium dendriticum in grazing beef cattle (NW Spain)

From February 2003 to March 2004, 1148 faecal samples from autochthonous Rubia Gallega cattle breed were collected in 170 farms from Lugo (NW Spain), in order to find out the prevalence and intensity of egg-excretion by Calicophoron daubneyi and Dicrocoelium dendriticum, and the possible influence of external factors as annual mean temperature and rainfall, altitude and mean slope. Twenty-six percent (95% CI 19%, 33%) and the 18% (12, 24) of the farms were infected with rumen and lancet flukes. C. daubneyi egg-output (Me=24) was found in 13% (11, 15) of the individual samples. Six percent (5, 7) of the animals in the study had low D. dendriticum eggs (Me=8). By estimating the odds ratio values it was shown that the highest probability of infection by C. daubneyi was in pastures with a mean slope of less than 13% (OR=1.9) and situated under 600 m (OR=1.6). Annual mean rainfall and temperature were not identified as risk factors for the infection with rumen flukes. Mountainous pastures with a mean slope of higher than 25% (OR=5.8) and situated over 600 m (OR=24.6) where precipitation was high (>1000 mm; OR=7) and temperature low (<11 degrees C; 2.8) had involved the highest risk of infection by D. dendriticum. Because of the prevalences found in this study, employment of suitable management practices with strategic treatments with efficacious anthelmintics are needed to reduce the presence of both trematodes and to increase the health status of grazing beef cattle.