Morphological diagnosis of infective larvae of Libyostrongylus douglassii (Cobbold, 1882) Lane, 1923 and L. dentatus Hoberg, Lloyd and Omar, 1995 (Nematoda: Trichostrongylidae) of ostriches

The differentiation of the species of the Libyostrongylus genus is only possible with the obtainment of the adult parasites in the ostriches proventriculus and gizzard. The present work confirms that it is possible to differentiate the infective larvae of L. douglassii and L. dentatus allowing the differential diagnosis of these species by fecal culture. To show this, adult females from both species were collected from ten proventriculus from adult ostriches and separated by species. Both groups were macerated individually added to sterilized feces for standard fecal cultures. The infective larvae were recovered, identified, quantified and measured. All proventriculus analyzed were parasitized by Libyostrongylus spp. and a clear heterogeneous location for each species was observed. The infective larvae from the fecal cultures of macerated L. douglassii presented a mean total length of 874.3+/-33.80 microm, and a short sheath tail (29.5+/-4.11 microm) with acute termination. The infective larvae from the macerated L. dentatus presented mean total length of 856.0+/-43.63 microm, long sheath tail (61.2+/-9.52 microm) with filamentous termination. The mean measures of the tails of both species had a significant difference. The differentiation of the infective larvae of L. douglassii and L. dentatus by fecal cultures will facilitate the diagnosis of both species for further understanding the Libyostrongylus biology.     

Isolation of a gammaherpesvirus similar to asinine herpesvirus-2 (AHV-2) from a mule and a survey of mules and donkeys for AHV-2 infection by real-time PCR

Equids are commonly infected by herpesviruses, but isolation of herpesviruses from mules has apparently not been previously reported. Furthermore, the genomic relationships among the various equid herpesviruses are poorly characterized. We describe the isolation and preliminary characterization of a mule gammaherpesvirus tentatively identified as asinine herpesvirus-2 (AHV-2; also designated equid herpesvirus-7 (EHV-7)) from the nasal secretions (NS) of a healthy mule in northern California. The virus was initially identified by transmission electron microscopic examination of lysates of cell culture inoculated with NS collected from the mule. A 913 nucleotide sequence of the DNA polymerase gene was amplified using degenerate primers, and comparison of this sequence with those of various other herpesviruses showed that the mule herpesvirus was most closely related to EHV-2 (AHV-2 sequences were not available for comparison). The sequence of a shorter portion (166 nucleotides) of the mule herpesvirus DNA polymerase gene was identical to that of the published sequence of an asinine gammaherpesvirus, previously designated as AHV-4-3 (AY054992). AHV-2 was detected by real-time polymerase chain reaction assay in the NS of approximately 8% of a cohort of 114 healthy mules and 13 donkeys.     

The involvement of mast cells and mast cell proteinases in the intestinal response to equine cyathostomin infection

Cyathostomins (Cyathostominae) are regarded as the most pathogenic equine nematode worldwide. These nematodes are difficult to control in equine populations due to emerging anthelmintic resistance and evasion of encysted larval cyathostomins to regular modern anthelmintics. Mast cells and their proteinases have been shown to play a role in the mammalian immune response to nematode infections. Involvement of mast cells and mast cell proteinases in the equine immune response to cyathostomin infection is proposed. A technique was established to perform immunohistochemical staining using polyclonal rabbit anti-equine mast cell proteinase-1 (eqMCP-1) and anti-equine tryptase on formalin-fixed large intestinal sections, from horses classified as cyathostomin positive and negative at the time of death based upon larval enumeration. Quantitative analysis of antibody labelled mast cells was used to detect mast cell proteinases in equine large intestinal sections positive and negative for cyathostomin larvae. This demonstrated an increase in equine tryptase labelled mucosal and submucosal mast cells in cyathostomin positive horses. This study has established an immunohistochemical technique to demonstrate mast cell proteinases in formalin-fixed large intestinal sections. This technique may be used to determine possible involvement of mast cells and their proteinases in the equine immune response to cyathostomin larvae. Further studies are required to define a specific role.     

A newly recognized blood group in domestic shorthair cats: the Mik red cell antigen

BACKGROUND: Naturally occurring alloantibodies produced against A and B red cell antigens in cats can cause acute hemolytic transfusion reactions. Blood incompatibilities, unrelated to the AB blood group system, have also been suspected after blood transfusions through routine crossmatch testing or as a result of hemolytic transfusion reactions. HYPOTHESIS: Incompatible crossmatch results among AB compatible cats signify the presence of a naturally occurring alloantibody against a newly identified blood antigen in a group of previously never transfused blood donor cats. The associated alloantibody is clinically important based upon a hemolytic transfusion reaction after inadvertent transfusion of red cells expressing this red cell antigen in a feline renal transplant recipient that lacks this red cell antigen. METHODS: Blood donor and nonblood donor cats were evaluated for the presence of auto- and alloantibodies using direct antiglobulin and crossmatch tests, respectively, and were blood typed for AB blood group status. Both standard tube and novel gel column techniques were used. RESULTS: Plasma from 3 of 65 cats and 1 feline renal transplant recipient caused incompatible crossmatch test results with AB compatible erythrocytes indicating these cats formed an alloantibody against a red cell antigen they lack, termed Mik. The 3 donors and the renal transplant recipient were crossmatch-compatible with one another. Tube and gel column crossmatch test results were similar. CONCLUSIONS AND CLINICAL IMPORTANCE: The absence of this novel Mik red cell antigen can be associated with naturally occurring anti-Mik alloantibodies and can elicit an acute hemolytic transfusion reaction after an AB-matched blood transfusion.     

Color and surface chemistry changes of extracted wood flour after heating at 120 °C

To investigate the effect of heat on color and surface chemistry of wood flour (WF), unextracted, extracted and delignified samples of commercial WF were heated at 120 °C for 24 h and analyzed by colorimetry, diffuse reflectance visible (DRV), attenuated total reflectance Fourier transform infrared (ATR-FTIR) and Fourier transform Raman (FT-Raman) spectroscopies. Unextracted samples showed a slight increase in CIEL^*a^*b^* color coordinates, a ^* and b ^*. Compared with unextracted samples, color changes of the extracted samples varied with composition of the extraction solvents with generally smaller increases in a ^* and larger decreases in b ^* values. Delignified samples were marked by even larger increases in both a ^* and b ^* values. The color changes could be explained by analysis of the respective DRV, FTIR and FT-Raman spectra of the samples before and after heating. Heating of the extracted WF at 120 °C resulted in a red shift of the absorption at 430 nm and increase in absorption in the violet-blue spectral region of visible light. Delignified samples showed even more pronounced absorption in this spectral region after heating.     

Use of Best Management Practices and Pasture and Soil Quality on Maryland Horse Farms

Agricultural operations, including horse farms, can contribute nonpoint source (NPS) pollution to surface water. The use of best management practices (BMPs) is the most effective way to prevent the movement of pollutants to surface water from nonpoint source pollution. Previous mailed survey studies have assessed the use of BMPs at the county and state level, but a visual assessment of horse farms is necessary to validate survey results. An observational field study was conducted to assess BMP use and soil and pasture quality and to create a model to predict soil erosion on Maryland horse farms. Fifty-one farms were selected based on stocking density (acres per horse [ac horse^-1]), farm use, and presence of water on property. All farms were visited from September through November 2009. In each pasture with grazing horses, the correct use of BMPs was assessed, grass height and vegetative cover were measured, and composite soil samples were collected. Less than half of the 18 assessed BMPs were being used by participants. Although most participants maintained the recommended vegetative cover and grass height, soil erosion was a major problem in pastures. Most farms had optimum soil nutrient concentrations (Ca, K, and P), excessive Mg values, and basic soil pH. Vegetative cover and grass height measurements were positively correlated with stocking density (r = 0.345, P < .0001; and r = 0.291, P < .0001, respectively). Farm use was the only variable that predicted soil erosion on farms (P = .006). Farms used for pleasure were least likely to have soil erosion, whereas farms used for breeding were more likely to have soil erosion (P = .0058). Despite the low-to-moderate adoption of BMPs, the maintenance of recommended vegetative cover and grass height as well as optimum values of soil nutrients indicated participating Maryland horse farms have a low potential for nutrient movement and NPS pollution.     

Individual vulnerability factors of Silver fir (Abies alba Mill.) to parasitism by two contrasting biotic agents: mistletoe (Viscum album L. ssp. abietis) and bark beetles (Coleoptera: Curculionidae: Scolytinae) during a decline process

? Context

In recent decades, there have been increasing reports of forest decline, especially in Mediterranean forest ecosystems. Decline in tree vitality is usually due to complex interactions between abiotic factors and biotic agents that attack weakened trees.

? Aims and methods

Estimating dendrometrical characteristics [basal area increment (BAI), age at DBH from tree ring counting, social status, height, and diameter], tree health status, and a competition index, we investigated the individual vulnerability of a French declining silver fir forest to both mistletoe (Viscum album L. ssp. abietis) and bark beetles (Pityophthorus pityographus Ratz., Pityokteines vorontzovi Jac., and Pityokteines spinidens Reitt.).

? Results

BAI was negatively correlated with both mistletoe infection (via mistletoe biomass) and bark beetle attack (number of insects per square meter), but there was evidence of divergence in tree choice between two groups of parasites. Mistletoe preferentially infected isolated and dominant trees that showed higher past growth rates than non-infected ones. Conversely, bark beetles mainly attacked defoliated and preferably declining trees with diameter (DBH) lower than 44.5 cm and slower past growth.

? Conclusion

While successive severe drought periods are thought to greatly weaken southern silver fir populations, mistletoe and bark beetles may contribute actively to their decline processes as inciting and contributing factors, respectively.     

Biofilm development within a larval rearing tank of the tropical rock lobster, Panulirus ornatus

The role of bacterial biofilms in disease processes is becoming increasingly recognised in both clinical and environmental settings. Biofilm development within a rearing tank of the tropical rock lobster Panulirus ornatus was studied to evaluate if the biofilm is a reservoir for potentially pathogenic bacteria that cause mass larval mortalities. Within a 5000 L larval rearing tank, fiberglass microscope slides were systematically distributed during a standard rearing attempt to assess biofilm development. Culture-based counts for two media types, TCBS and Marine Agar (MA), demonstrated increased bacterial densities until days 11 and 13 respectively. For both media types, a drop in the plate counts was followed by a subsequent increase towards the end of the experiment. Scanning electron microscopy (SEM) confirmed that cell densities decreased between days 13 and 17, most likely due to sloughing of the biofilm into the water column. SEM images revealed distinct changes in dominant morphologies reflecting a succession of bacterial populations. A dynamic succession of microbial species during biofilm development was also demonstrated using denaturing gradient gel electrophoresis (DGGE) profiling of bacterial 16S rRNA genes in combination with statistical ordination analysis. Prominent changes in the DGGE profiles coincided with the decrease in bacterial numbers observed by SEM and plating on MA between days 13 and 17. Fluorescence in situ hybridization (FISH) identified -Proteobacteria as being numerically abundant in the biofilm. This was supported by results from DGGE analysis, which retrieved only sequences affiliated with - and γ-Proteobacteria. DGGE bands affiliated with Vibrio became dominant towards the end of the larval run (days 21 to 24). A Vibrio harveyi strain isolated from the biofilm late in the larval rearing trial (day 24) demonstrated increased larval mortality in small scale phyllosoma survival studies. The detection of Vibrionaceae at the end of the larval trial coincided with mass phyllosoma mortality and show that the biofilm is a reservoir for potentially pathogenic bacteria.