Microscopic examination of the intestinal wall and selected organs of minipigs orally supplemented with humic acids

Humic acids are used to prophylactically treat intestinal diseases in a wide number of species, yet the mechanism of action remains unknown. The general assumption has been that humic acids act locally; however studies using young piglets show orally supplemented humic acids can penetrate the intestinal wall, and thus potentially act systemically. The objective of this study was to determine if humic acids could also cross the intestinal barrier in adult pigs and be detected in other organs. Adult minipigs (>18 months old) orally received either 1 g humic acids/kg body weight (verum, n = 3) or placebo (control, n = 3), for 2 weeks. At the end of the feeding period tissue samples were harvested from the intestine, various glands and organs. Unstained tissue samples were examined by light microscopy for the presence of humic acid particles. No humic acid particles were detected in any of the unstained tissues from verum or control pigs.     

Marketing of meat sheep with intensive finishing in southern state of Mexico

The objective of the present study was to quantify the marketing margins of cold sheep carcass and barbacoa meat in the southern state of Mexico. Data were obtained from the production chain links (production, industrialization, and marketing) where the marketing margins were calculated along with the benefit/cost (B/C) ratio. In the absolute marketing margin of the final consumer price per kilogram of carcass meat, the producer obtained US$2.7/kg (47 %) of the utilities, while the intermediaries obtained US$3.1/kg (53 %). Considering the final cooked product in barbacoa (typical dish), the margin was US$6.3/kg (29 %) for the producer and US$15.2/kg (71 %) for the intermediaries. The B/C ratio was 1.0, 1.1, 2, and 1.3 for the producer, stocker, butcher, and barbacoa seller, respectively. It is concluded that the best marketing channel for the producer was the producer-stocker-processor (butcher and barbacoa seller). The highest marketing margin was for the intermediaries followed by the producer. The order of importance of the B/C kilogram ratio of meat was for the butcher first, then barbacoa seller, and lastly stocker and producer.     

Putative human and avian risk factors for avian influenza virus infections in backyard poultry in Egypt

Highly pathogenic influenza A virus subtype H5N1 causes significant poultry mortality in the six countries where it is endemic and can also infect humans. Egypt has reported the third highest number of poultry outbreaks (n = 1084) globally. The objective of this cross-sectional study was to identify putative risk factors for H5N1 infections in backyard poultry in 16 villages in Damietta, El Gharbia, Fayoum, and Menofia governorates from 2010–2012. Cloacal and tracheal swabs and serum samples from domestic (n = 1242) and wild birds (n = 807) were tested for H5N1 via RT-PCR and hemagglutination inhibition, respectively. We measured poultry rearing practices with questionnaires (n = 306 households) and contact rates among domestic and wild bird species with scan sampling. Domestic birds (chickens, ducks, and geese, n = 51) in three governorates tested positive for H5N1 by PCR or serology. A regression model identified a significant correlation between H5N1 in poultry and the practice of disposing of dead poultry and poultry feces in the garbage (F = 15.7, p < 0.0001). In addition, contact between domestic and wild birds was more frequent in villages where we detected H5N1 in backyard flocks (F = 29.5, p < 0.0001).     

Clodronate treatment significantly depletes macrophages in chickens

Macrophages function as phagocytes and antigen-presenting cells in the body. As has been demonstrated in mammals, administration of clodronate [dichloromethylene bisphosphonate (Cl₂MBP)] encapsulated liposomes results in depletion of macrophages. Although this compound has been used in chickens, its effectiveness in depleting macrophages has yet to be fully determined. Here, we show that a single administration of clodronate liposomes to chickens results in a significant depletion of macrophages within the spleen and lungs of chickens up to 4 d post-treatment. This finding suggests that, in order to obtain depletion of macrophages in chickens for greater than 5 d, it is necessary to administer clodronate liposomes 4 d apart. The study also showed that 2 treatments of clodronate liposomes at 4-day intervals resulted in the depletion of macrophages for up to 10 d. The findings of the present study will encourage more precise studies to be done on the potential roles of macrophages in immune responses and in the pathogenesis of microbial infections in chickens.     

In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

Fungal inoculation induces agarwood in young Aquilaria malaccensis trees in the nursery

Fungi are often used to induce agarwood inAquilariatrees. This study was conducted to evaluate the effect of several fungi on agarwood formation over time in youngAquilaria malaccensis （Lam.） trees. Typical changes in the length and light intensity of the resulting discoloration were observed after three and six month periods following inoculation. Wood samples were observed microscopically and classified into several light intensity groups. The discoloration length was meas-ured longitudinally. The duration after inoculation affected the mean of discoloration length： the 6-month old sample （1.70 cm） had a wider discoloration zone when compared to the 3-month old sample （1.17 cm）. When measuring the discoloration intensity, a positive relationship with time was perceived. Digital images, captured using a camera-equipped microscope, revealed that wood samples collected after six months appeared to be 1.8-times darker than after three months. We concluded that time, not the species of any of the tested fungi, had significant effect on discoloration length and intensity. Gas chromatography/mass spec-trometry （GCMS） analysis of the 6-month old sample yielded some important agarwood compounds such as benzylacetone, anisylacetone, guaiene and palustrol. This demonstrates that the tested fungi have the ability to induce agarwood formation in nurseryA. malaccensis trees.     

Early weaning of winter flounder (Pseudopleuronectes americanus Walbaum) larvae on a commercial microencapsulated diet

Like most small marine fish larvae, the stomachs of winter flounder Pseudopleuronectes americanus are undeveloped at first feeding and have relatively reduced digestive capacity. This work was undertaken to test whether larvae at the onset of stomach differentiation (larval size about 5.5 mm) could be early weaned onto a commercial microencapsulated diet. We assessed the effect of early weaning by first comparing growth performance (standard length, total protein content and age at metamorphosis) of larvae fed enriched live prey from first feeding to a size of 5.5 mm and then reared on three different feeding regimes until metamorphosis: (1) live prey (LP) as a control group; (2) mixed feeding of live prey and microencapsulated diet (LP‐ME); (3) exclusively microencapsulated diet (ME) after fast weaning over 4 days (to a larval size of 6.2 mm). No differences were observed between larval development in the two first groups, which began metamorphosis at 40 days old. The larvae of the third group showed significantly slower growth that resulted in a delay of 4 days in the onset of metamorphosis. Differences in live prey availability between the treatments and the short transition period to allow the larvae to adapt to the new diet were identified as possible contributing factors to the slower growth and to the delay in metamorphosis of early weaned larvae. In a second experiment, the transitional weaning period was increased until the larvae were 6.6 mm in length. Weaning at that size resulted in no slowing of growth or delay in metamorphosis, suggesting that the feeding schedule was adequate.     

Equine hyperimmune serum protects mice against Clostridium difficile spore challenge

Clostridium (C.) difficile is a common cause of nosocomial diarrhea in horses. Vancomycin and metronidazole have been used as standard treatments but are only moderately effective, which highlights the need for a novel alternative therapy. In the current study, we prepared antiserum of equine origin against both C. difficile toxins A and B as well as whole-cell bacteria. The toxin-neutralizing activities of the antibodies were evaluated in vitro and the prophylactic effects of in vivo passive immunotherapy were demonstrated using a conventional mouse model. The data demonstrated that immunized horses generated antibodies against both toxins A and B that possessed toxin-neutralizing activity. Additionally, mice treated with the antiserum lost less weight without any sign of illness and regained weight back to a normal range more rapidly compared to the control group when challenged orally with 10⁷ C. difficile spores 1 day after serum injection. These results indicate that intravenous delivery of hyperimmune serum can protect animals from C. difficile challenge in a dose-dependent manner. Hence, immunotherapy may be a promising prophylactic strategy for preventing C. difficile infection in horses.     

Growth performance and feed utilization of Nile tilapia Oreochromis niloticus (Linnaeus, 1758) and tilapia galilae Sarotherodon galilaeus (Linnaeus, 1758) fingerlings fed plant protein-based diets

This study was designed to determine the effect of complete substitution of fish meal (FM) by three plant protein sources including extruded soybean meal (SBM), extruded full‐fat soybean (FFSB) and corn gluten meal (CGM) on growth and feed utilization of Nile tilapia Oreochromis niloticus and tilapia galilae Sarothrodon galilaeus. Four isonitrogenous of crude protein (ca. 28.0%) and isocaloric (ca. 19 MJ kg⁻¹) experimental diets were formulated. The control diet (diet 1) was prepared with FM as the main protein sources. Diets 2–4, each FM control diet, were completely substituted with SBM (diet 2), FFSB (diet 3) and CGM (diet 4). l ‐lysine and dl ‐methionine were added to plant protein diets to cover the nutritional requirements of tilapia. Each treatment was allocated to three net pens and fed for 17 weeks. Nile tilapia fed the control diet showed significantly higher (P≤0.05) values for final body weight (FBW), feed intake (FI), weight gain (WG) and specific growth rate (SGR), whereas fish fed the diet with CGM achieved the lowest values. Tilapia galilae fed SBM diet recorded the highest (P≤0.05) values for growth performance. Better feed conversion ratio (FCR) for both Oreochromis niloticus and Sarothrodon galilaeus was observed when fish were fed SBM diet, whereas the worse FCR was recorded for FFSB diet. Feed utilization parameters including protein productive value (PPV), fat retention (FR) and energy retention (ER) showed significant differences (P≤0.05) for both the species fed different dietary protein sources. The present results suggest that, for Nile tilapia, both SBM and FFSB supplemented with dl ‐methionine and l ‐lysine can completely replace dietary FM. Meanwhile, S. galilaeus fed SBM diet exhibited comparable growth and feed utilization with those fish fed a fish‐meal‐based diet.