Assessment of in vitro interferon-gamma responses from peripheral blood mononuclear cells of cattle infected with Mycoplasma mycoides ssp. mycoides small colony type

Contagious bovine pleuropneumonia (CBPP) is a lung disease caused by the bacterial pathogen Mycoplasma mycoides ssp. mycoides small colony type (MmmSC). It has been spreading due to a number of factors including poor vaccine efficacy and poor sensitivity of current diagnostic tests. The purpose of this study was to assess interferon gamma (IFN-gamma) release after stimulation of peripheral blood mononuclear cells (PBMC) from experimentally infected cattle. PBMC collected from 15 artificially infected animals were incubated with different concentrations of total MmmSC antigen. After 72h of incubation the IFN-gamma release was measured and found to be elevated in 11 animals. We did not observe a correlation between IFN-gamma release of animals with and without pathomorphological gross lesions. Therefore, our data do not confirm a role for CD4 T-lymphocytes in protection, since there is no correlation between IFN-g secretion (supposed to be mainly derived from CD4 T-cells) and disease severity. Additionally, we applied immunocytochemistry on affected lung tissue and detected no build up of T-lymphocytes (CD4 T-cells, CD8 T-cells) but a high presence of myeloid cells.     

Use of dried blood smears for detection of feline hemoplasmas using real-time polymerase chain reaction

The objective of the current study was to determine the sensitivity and specificity of real-time polymerase chain reaction (real-time PCR) for feline hemoplasmas when applied to DNA extracted from dried whole-blood smears in comparison to that for DNA extracted from liquid whole blood. Blood samples were collected into ethylenediamine tetra-acetic acid tubes from 305 cats with possible or suspected hemoplasmosis, and dried blood smears from each sample were prepared. DNA was extracted from blood smears and a 160-microl aliquot of each liquid blood sample by using a robotic extractor and was subjected to real-time PCR for feline glyceraldehyde-3-phosphate dehydrogenase (liquid blood), 18S ribosomal RNA (dried blood), and "Candidatus Mycoplasma haemominutum", Mycoplasma haemofelis, and "Candidatus Mycoplasma turicensis" DNA. When using the results for liquid whole blood as the gold standard, the sensitivity of each assay for "Ca. M. haemominutum", M. haemofelis, and "Ca. M. turicensis" was 49 of 66 (74%), 11 of 13 (85%), and 11 of 20 (55%), respectively. The specificity of each assay was 224 of 234 (96%), 287 of 287 (100%), and 280 of 280 (100%), respectively. When possible, liquid blood samples should be submitted for detection of feline hemoplasmas by using real-time PCR. The improved sensitivity of real-time PCR on blood smears for M. haemofelis compared with that of the other hemoplasma species may reflect the higher organism burdens associated with infection with this species.     

An investigation of Leishmania spp. in Didelphis spp. from urban and peri-urban areas in Bauru (São Paulo, Brazil)

Blood and bone marrow samples were taken from 112 Didelphis spp., collected between March 2005 and February 2006, from urban and peri-urban areas of Bauru, São Paulo State, Brazil, to evaluate the hypothesis that these animals might constitute a reservoir of Leishmania spp. Anti-Leishmania ssp. antibodies were screened in the serum samples using an enzyme-linked immuno-sorbent assay (ELISA) and the polymerase chain reaction (PCR). PCR was performed on fragments of DNA samples from Leishmania spp. using primers 13A and 13B, and showed a positive outcome in 91.6% of the 112 samples tested. Of the 107 samples analyzed by ELISA, 71% were positive. Evidence of epidemiological risk factors such as a circulating parasite and freely moving vectors suggests that Didelphis spp. may participate in the transmission cycle of Leishmania spp. in Bauru.     

Preparation of Extruded Snacks with Flavored Flour Obtained from the Carcasses of Nile Tilapia: Physicochemical,Sensory, and Microbiological Analysis

This study aimed to develop extruded snacks including flour obtained from Nile tilápia carcasses, and then evaluate the chemical composition, colorimetry, thiobarbituric acid reactive substances, and microbiology. There was a significant increase in the levels of crude protein, ether extract, ash, and minerals, as well as a reduction in carbohydrates as more flour was included in the snacks. Increased levels of flavored flour caused reduction in the brightness of snacks. Oxidative rancidity was not detected in snacks. The inclusion of flour improved the nutritional value of extruded snacks without changing sensory characteristics. Microbiologically, the snacks are suitable for human consumption.     

Extrusion of Cross‐Linked Hydroxypropylated Corn Starches II. Morphological and Molecular Characterization

A series of cross‐linked hydroxypropylated corn starches were extruded with a Leistritz micro‐18 co‐rotating extruder. Extrusion process variables including moisture (30, 35, and 40%), barrel temperature (60, 80, and 100°C), and screw design (low, medium, and high shear) were investigated. Scanning electron microscopy (SEM) of extruded starches showed a gel phase with distorted granules and granule fragments after extrusion at 60°C. After extrusion at 100°C only a gel phase was observed with no granular structures remaining. High performance size exclusion chromatography (HPSEC) equipped with multiangle laser light‐scattering (MALLS) and refractive index (RI) detectors showed extruded starches degraded to different extents, depending on extrusion conditions. The average molecular weight of the amylopectin of unextruded native corn starch was 7.7 × 10⁸. Extrusion at 30% moisture, 100°C, and high shear reduced the molecular weight of amylopectin to 1.0 × 10⁸. Hydroxypropylated normal corn starch extruded at identical conditions showed greater decreases in amylopectin molecular weight. With the addition of cross‐linking, the amylopectin fractions of the extruded starches were less degraded than those of their native and hydroxypropylated corn starch counterparts. Similarly, increasing moisture content during extrusion lowered amylopectin degradation in the extruded starches. Increasing temperature during extrusion of cross‐linked hydroxypropylated starches at high moisture content (e.g., 40%) lowered amylopectin molecular weights of the extruded starches, whereas increasing extrusion temperature at low moisture content (30%) resulted in less degraded molecules. This difference was attributed to the higher glass transition temperatures of the cross‐linked starches.     

Rapid sequence modification in the highly polymorphic region (HPR) of the hemagglutinin gene of the infectious salmon anaemia virus (ISAV) suggests intra-segmental template switching recombination

The ISAV has a genome composed of eight segments of (–)ssRNA, segment 6 codes for the hemagglutinin–esterase protein, and has the most variable region of the genome, the highly polymorphic region (HPR), which is unique among orthomyxoviruses. The HPR has been associated with virulence, infectivity and pathogenicity. The full length of the HPR is called HPR0 and the strain with this HPR is avirulent, in contrast to strains with deleted HPR that are virulent to varying degrees. The molecular mechanism that gives rise to the different HPRs remains unclear. Here, we studied in vitro the evolution of reassortant recombinant ISAV (rISAV) in Atlantic salmon head kidney (ASK) cells. To this end, we rescued and cultivated a set of rISAV with different segment 6-HPR genotypes using a reverse genetics system and then sequencing HPR regions of the viruses. Our results show rapid multiple recombination events in ISAV, with sequence insertions and deletions in the HPR, indicating a dynamic process. Inserted sequences can be found in four segments of the ISAV genome (segments 1, 5, 6, and 8). The results suggest intra-segmental heterologous recombination, probably by class I and class II template switching, similar to the proposed segment 5 recombination mechanism.     

Using acoustic telemetry and snorkel surveys to study diel activity and seasonal migration of round goby (Neogobius melanostomus) in an estuary of the Western Baltic Sea

In just two decades, round goby Neogobius melanostomus (Pallas) has dispersed throughout most of the Baltic and the North American Great Lakes. It thrives in shallow sheltered areas, where it is impacting native fauna. In Denmark, researchers, governmental institutions and fishers have joined in an effort to establish a sustainable round goby fishery. To do so, basic information about the movement of round goby is essential, so the movements of 50 round gobies were tracked using acoustic telemetry. The results revealed that activity took place mainly at night and seasonal migrations were common. The most frequent migration behaviour observed was a seaward return migration during winter, which was further confirmed by a snorkel survey. Tracking data also revealed riverine upstream dispersal. The results demonstrate that the round goby can be mapped using telemetry, and it is suggested that fishing for round goby may improve efficiency by targeting seasonal migration corridors.     

Climate effects and bottom‐up controls on growth and size‐at‐age of Pacific halibut (Hippoglossus stenolepis) in Alaska (USA)

Pacific halibut (Hippoglossus stenolepis) are an ecologically, commercially, and culturally important Alaskan groundfish species. Commercial harvest of halibut dates back to the late 19th century and has been managed by the International Pacific Halibut Commission (IPHC) since 1921. IPHC surveys have revealed declining trends in survey biomass in multiple regions and region‐specific declines in mean size‐at‐age (size‐at‐age) over the past two decades (>50% in some areas). Changes in size‐at‐age can arise from a variety of physical, ecological, sampling, and fishery effects, including size‐dependent fishery or predation mortality, alteration in growth from variability in prey quality or quantity, and changes in temperature‐dependent metabolic demands. Here, we develop and apply a bioenergetics model for halibut using survey‐based diet and temperature data for Alaska to evaluate potential environmental drivers of size‐at‐age. In general, juvenile (<40 cm fork length) foraging rates were highest in the Gulf of Alaska concomitant with higher potential growth and elevated basal metabolic demands during warm summer conditions. In contrast, adult (40–120 cm FL) potential growth was highest in the Eastern Bering Sea, potentially reflecting lower metabolic costs and higher rates of prey consumption in that region. We additionally find evidence for interannual variation in potential growth, with a higher frequency of reduced growth potential in the last decade, particularly in the Eastern Bering Sea in 2015 and 2016 for both juvenile and adult halibut. These results suggest the potential for patterns in size‐at‐age to arise from trophic and environmental constraints that collectively limit growth in some regions and years.