Linoleic acid increases adhesion,chemotaxis, granule release,intracellular calcium mobilisation,MAPK phosphorylation and gene expression in bovine neutrophils

Neutrophils are critical to the innate immune response; therefore, the proper function of neutrophils is critical to avoid the development of certain diseases. Linoleic acid, a polyunsaturated long-chain fatty acid, is one of the most abundant long-chain fatty acids found in the plasma of cows after giving birth. In this study, we evaluated the effects of linoleic acid treatment on bovine neutrophil adhesion, chemotaxis, metalloproteinase (MMP)-9 release, CD11b expression, intracellular calcium mobilisation, mitogen-activating protein kinase (MAPK) phosphorylation and COX-2 and IL-8 expression. Bovine neutrophils isolated from healthy heifers were incubated with different concentrations of linoleic acid, and then neutrophil responses were evaluated. Our results show that the treatment of neutrophils with 100 μM linoleic acid increased their adhesion to the bovine endothelial cell line CPA47. The results of a transwell migration assay revealed that linoleic acid could also promote the chemotaxis of bovine neutrophils. Furthermore, linoleic acid treatment increased MMP-9 activity and CD11b cell surface expression in neutrophils. Fifty and 100 μM linoleic acid also increased intracellular calcium mobilisation in neutrophils loaded with Fluo-4 AM dye. Linoleic acid also rapidly (2–5 min) stimulated the phosphorylation of ERK1/2 and p38 MAPK as evaluated by immunoblot. Finally, COX-2 and IL-8 mRNA expression increased after 2 h of linoleic acid treatment. In conclusion, linoleic acid stimulates adhesion, chemotaxis, granule release and intracellular responses in bovine neutrophils.     

Antimicrobial susceptibility patterns of Haemophilus parasuis from pigs in the United Kingdom and Spain

A total of 30 British and 30 Spanish Haemophilus parasuis isolates were tested for their susceptibility to 19 of the antimicrobials currently used in swine practice with a broth microdilution method in order to know the emergence of resistance against these compounds in this porcine pathogen. All the British isolates were susceptible to penicillin, ceftiofur, erythromycin, tilmicosin, enrofloxacin, and florfenicol, and most of them were susceptible to the remaining antimicrobials (the highest resistance rate found was of 20% to neomycin). In contrast, all the Spanish isolates were susceptible exclusively to florfenicol, and high proportions of resistance were encountered for penicillin, ampicillin, oxytetracycline, erythromycin, tilmicosin, tiamulin and trimethoprim+sulphamethoxazole; in addition, a bimodal or multimodal distribution, or tailing of Spanish isolates over the MIC range was observed for clindamycin, sulphonamides and tylosine tartrate, suggesting the development of acquired resistance. In addition, several multiresistance patterns were found among the Spanish isolates, 23.3% of them being resistant to at least eight antimicrobials, the same rate as that encountered for those being susceptible to all antimicrobials tested. This study showed that in general British H. parasuis isolates are susceptible to antimicrobial agents routinely used for treatment of porcine respiratory diseases; however, the Spanish isolates need a more continuous surveillance of their susceptibility patterns.     

Ileal digestibility of protein and amino acids from canola meal in weaned piglets and growing pigs

Two experiments were conducted to measure the apparent ileal digestibility coefficients (AID) of protein and amino acids from canola meal (CM) and a pelleted canola meal (PCM) and their effect on specific activity (SA) of pancreatic proteases in weaned piglets and growing pigs. In experiment one, 24 piglets weaned at 17 days and weighing 5.5 kg were used. Treatments were a reference diet with 200 g of crude protein (CP) kg^− 1 elaborated with casein (C) as the sole protein source, a diet containing C–CM and a diet containing C–PCM. These diets were obtained using the reference diet plus 100 g kg^− 1 of CM or PCM that substituted an isoproteic mixture of casein and maize starch from reference diet, so that the AID coefficients for their protein and amino acids could be calculated by difference. In experiment two, nine castrated pigs weighing 39.5 kg were used. Treatments were a reference diet with 160 g of CP kg^− 1 elaborated with casein (C) as the sole protein source, diet C–CM and diet C–PCM. These diets were obtained using the reference diet plus 300 g kg^− 1 of CM or PCM that substituted an isoproteic mixture of casein and maize starch from reference diet. In piglets, the AID coefficients for casein were highest (P < 0.05), those of PCM were intermediate, and those of CM were the lowest. In older pigs, the AID coefficients for casein were highest, and those of CM and PCM were similar (P > 0.05). The SAs of chymotrypsin, trypsin and carboxypeptidases A and B were lower in piglets than in heavier pigs. Moreover, the SAs of trypsin, chymotrypsin, and carboxypeptidase B were lower (P > 0.05) in animals fed casein. The results showed in piglets that whereas CM was less digestible, pelleted canola meal improved protein and amino acid ileal digestibility, resulting in similar AID coefficients to those of growing pigs.     

Usefulness of the murine model to study the immune response against Histoplasma capsulatum infection

The present paper is an overview of the primary events that are associated with the histoplasmosis immune response in the murine model. Valuable data that have been recorded in the scientific literature have contributed to an improved understanding of the clinical course of this systemic mycosis, which is caused by the dimorphic fungus Histoplasma capsulatum. Data must be analyzed carefully, given that misinterpretation could be generated because most of the available information is based on experimental host–parasite interactions that used inappropriate proceedings, i.e., the non-natural route of infection with the parasitic and virulent fungal yeast-phase, which is not the usual infective phase of the etiological agent of this mycosis.     

Evidence for widespread Leishmania infantum infection among wild carnivores in L. infantum periendemic northern Spain

Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish Basque Country (BC), by PCR and DNA sequencing. The region is at the northern periphery of Leishmania infantum endemic Iberian Peninsula and infection in the dog (reservoir) or other species has not been previously reported. Leishmania kinetoplast DNA was detected by real-time PCR (rtPCR) in 28% (44/156) of animals. Specifically, in 26% of Eurasian badgers (n = 53), 29% of foxes (n = 48), 29% of stone martens (n = 21) and in 25–50% of less numerous species including genets, wild cats, pole cats, European mink and weasels. Infected animals particularly badgers, were most prevalent in the southernmost province of the BC (Araba) in areas dominated by arable land. Subsequent amplification and sequencing of a fragment of the rRNA internal transcribed spacer 2 (ITS2) from a subset of rtPCR positives samples confirmed the species as L. infantum, showing a high sequence homogeneity with ITS2 sequences of L. infantum from dogs and humans from southern Spain. In summary, this study reports for the first time L. infantum infection in wild carnivores from the BC including in stone martens, pole cats and minks in which infection has not been previously described. It supports the need to study infection in dogs and people in this region and is an example of the value of infection surveillance in wildlife to assess potential risks in the domestic environment and their role in spreading infections in non-endemic areas.     

Combined effect of using near‐infrared spectroscopy for nutritional evaluation of feed ingredients and non‐starch polysaccharide carbohydrase complex on performance of broiler chickens

This study was carried out to evaluate the combined effect of using near‐infrared spectroscopy (NIRS ) for nutritional evaluation of feed ingredients and the addition of non‐starch polysaccharide carbohydrase complex (NSP enzymes) on the growth performance of broilers fed diets produced with low‐quality wheat and soybean meal. A 2 × 2 trial design was performed, with seven replicates of 40 male Ross 308 broilers per treatment, evaluating the effect of the addition of NSP enzymes and the ingredients’ nutritional matrix based on table values or NIRS values. Diets without added enzymes were formulated to reach nutritional requirements, whereas diets with enzymes were reformulated, reducing the apparent metabolizable energy (AME ) by 85 kcal/kg. In the overall period (days 0–35), broilers fed diets formulated using NIRS values had higher (P  < 0.001) average daily gain (+11.3%) and daily feed intake (+7.2%), and a lower (P  < 0.001) feed conversion ratio (?5.3%) compared to those fed diets formulated using table values. When formulating diets for broilers with low‐quality feed ingredients, performance can be improved by considering NIRS values and by the addition of NSP enzymes, even with a reduction of AME . These nutritional approaches are efficient in improving broilers’ performances by themselves and even more so when they are combined.     

Histopathological and parasitological study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum

Background

The aim of this study was to provide a systematic pathological and parasitological overview of the gastrointestinal tract (GIT), including the stomach, duodenum, jejunum, ileum, caecum and colon, of dogs naturally infected with Leishmania.

Methods

Twenty mongrel dogs naturally infected with Leishmania (Leishmania) infantum and obtained from the Control Zoonosis Center of the Municipality of Ribeirão das Neves, Belo Horizonte Metropolitan area, Minas Gerais (MG) state, Brazil, were analyzed. The dogs were divided into two groups: Group 1 comprised nine clinically normal dogs and group 2 comprised 11 clinically affected dogs. After necropsy, one sample was collected from each GIT segment, namely the stomach, duodenum, jejunum, ileum, caecum and colon. Furthermore, paraffin-embedded samples were used for histological and parasitological (immunohistochemistry) evaluation and a morphometrical study were carried out to determine the parasite load (immunolabeled amastigote forms of Leishmania). The Friedman and the Mann Whitney tests were used for statistical analysis. The Friedman test was used to analyze each segment of the GIT within each group of dogs and the Mann Whitney test was used to compare the GIT segments between clinically unaffected and affected dogs.

Results

The infected dogs had an increased number of macrophages, plasma cells and lymphocytes, but lesions were generally mild. Parasite distribution in the GIT was evident in all intestinal segments and layers of the intestinal wall (mucosal, muscular and submucosal) irrespective of the clinical status of the dogs. However, the parasite load was statistically higher in the caecum and colon than in other segments of the GIT.

Conclusion

The high parasite burden evident throughout the GIT mucosa with only mild pathological alterations led us to consider whether Leishmania gains an advantage from the intestinal immunoregulatory response (immunological tolerance).     

Nutritional value of sugarcane silage enriched with corn grain,urea, and minerals as feed supplement on growth performance of beef steers grazing stargrass

The objectives were to evaluate the silage quality of sugarcane silage enriched (as fed) with corn grain ground (10%), urea (1.5%), and mineral premix (0.5%) and its effects as a feed supplement on growth performance of beef steers grazing stargrass. Firstly, in micro-silages, whole sugarcane enriched with corn grain, urea, and minerals (WSCE) were ensilaged by 0, 20, 40, and 60 days. Crude protein (CP) and lactic acid (LA) increased linearly (P < 0.05) and true protein decreased linearly (P < 0.05) as fermentation time increased. The pH values in silages were affected quadratically by fermentation time. Thus, after a 20-day fermentation, the pH values were below 4.5. Secondly, in micro-silos WSCE and stem sugarcane enriched with corn grain, urea, and minerals (SSCE) with and without calcium propionate were ensilaged by 30 days, but it did not affect any chemical composition trait in the silage. The SCCE silages had higher CP and LA and lower pH than WSCE silages. Finally, for 120 days, 20 beef steers (378 ± 33 kg initial BW) grazing stargrass were supplemented (daily by 1-h free access) with WSCE silage. Supplemental silage increased total dry matter intake, total gain, and the average daily gain, without any affectation on feed conversion and total tract digestion of dry matter. It is concluded that whole sugarcane silage is an alternative feed supplement to improve growth performance in beef steers grazing stargrass.