Introduction: There is still lack of morphological and phylogenetic information on the pathogenic nematode of the camel Haemonchus longistipes. In the present study, this parasite was isolated in Saudi Arabia and described. Material and Methods: The abomasa of two Arabian camels were collected from a slaughterhouse in Abha province and examined for nematode infection. Worms were described morphologically and morphometrically by electron microscopy. Multiple sequence alignment and the phylogenetic tree of the parasite were constructed from maximum likelihood analysis of its ITS-2 rDNA sequences. Results: These nematodes had a slender body terminating anteriorly at a conspicuous dorsal lancet. A pair of lateral cervical papillae distant from the anterior end was observed. The buccal aperture was hexagonal and surrounded by two amphids, six externo-labial papillae, and four cephalic papillae. Males terminated posteriorly at a bursa supported by spicules and lateral and dorsal rays. Females were linguiform and knobbed morphotypes with distinct ovijectors and a dorsal rim covering the anal pore. The taxonomy was confirmed by the morphology and number of the longitudinal cuticular ridges in a 43–46 range. The sequence alignment and phylogeny revealed 92% homology with H. longistipes (AJ577461.1), and the sequence was deposited into GenBank. Conclusion: The present study describes H. longistipes morphologically and molecularly which facilitates further discrimination of this species worldwide.

Two cytopathic agents were isolated on porcine alveolar macrophages following inoculation with homogenates of lung tissues from pigs showing respiratory problems. These isolates were identified as porcine reproductive and respiratory syndrome (PRRS) virus isolates by indirect immunofluorescence using a PRRS virus (PRRSV) specific monoclonal antibody (MAb) and were designated as LHVA-92-1 and LHVA-92-2. Immunogold electron microscopy using a porcine PRRS positive serum pool and protein A-gold resulted in an intense labelling of aggregates of viral particles. Dark specific cytoplasmic staining of porcine alveolar macrophages infected with both virus isolates could be observed by immunogold silver staining (IGSS) using the specific MAb. This method proved effective in detecting PRRSV antigens in several ethanol-fixed tissues of piglets intranasally inoculated with the supernatants of macrophages infected with each isolate. Immunogold silver staining was also successfully used for the detection of PRRSV antigens on sections of formalin-fixed paraffin-embedded lung tissues and on frozen sections of lungs. The present results indicate that colloidal gold may be useful for the identification and immunohistochemical detection of PRRSV in tissues.

A double-antibody ELISA for the detection of coronaviruses in intestinal contents from turkey poults with diarrhea was developed. Antibodies were raised in rabbits and guinea pigs against a Minnesota isolate of turkey enteric coronavirus (TCV) propagated in embryonating turkey eggs and were purified by density-gradient centrifugation. The specificity of antisera was confirmed by hemagglutination-inhibition and immunoelectron microscopy. Absorption of anti-TCV hyperimmune sera with egg extracts or egg ovalbumin and the use of different dilution and blocking buffers influenced the sensitivity and specificity of the ELISA. Reciprocal cross-reactivity was detected among turkey, chicken, bovine, and murine coronaviruses. Antisera to the transmissible gastroenteritis virus of swine, the rabbit enteric coronavirus, or the human coronavirus strain 299E failed to react with TCV. The TCV cross-related only moderately with the avian infectious bronchitis virus and the hemagglutinating encephalomyelitis virus of swine. Investigations with samples from 47 commercial turkey flocks in Quebec with episodes of transmissible enteritis revealed that the ELISA was more sensitive than was electron microscopy for dectection of TCV.

Electron microscopy revealed several unique features in canine bone marrow, compared with that of other species. The marrow was fatty and extensively trabeculated and was enclosed by a complete layer of endosteal bone-lining cells. Branched reticular cells were closely associated with each other and, occasionally covered part of the sinus wall as an adventitial layer. The extent of adventitial coverage varied markedly and was less extensive, compared with that of other species. On average, only 23% of the sinus wall was covered by adventitial layer, in contrast to 65% reported in laboratory animals. Unilaminar sinuses, with no adventitial coverage, accounted for greater than 38% of all sinuses. Quantitative analysis indicated that 60% of the latter sinuses contained apertures, as opposed to 35% of sinuses with adventitial coverage (P less than 0.05). Moreover, the number of apertures in unilaminar sinuses was significantly (P less than 0.009) greater than that in multilaminar sinuses. Apertures were observed every 59 micromoles in unilaminar sinuses, in contrast to every 109 micromoles in multilaminar sinuses. Approximately 75% of the apertures were occupied by cells in transit, and only 25% were free of cells. Macrophages were distributed throughout the marrow and were closely associated with all blood cell lines. Occasionally, cells that entered the lumen were not fully mature. Erythroblasts were seen migrating across the wall and within the lumen of sinuses. The less-extensive adventitial coverage in canine bone marrow might indicate that the rate of cell delivery from the marrow into the circulation was relatively high in this species. The prevalence of unilaminar sinuses, along with the larger number of apertures, suggested that these sinuses were more accessible to the migrating cells and that the cellular traffic across them was intense.

FowlPox virus (FPV) was detected in eight chickens suffering from a diphtheritic lesion on the oropharynx and trachea with nodular skin lesions around the unfeathered parts in two Egyptian governorates (El-Sharkia and Ismailia governorates) during summer 2023. A variety of serological and molecular methods were performed for identification and characterization of the virus. on specific-pathogen free (SPF) embryonated chicken eggs via chorio-allantoic membranes (CAM), the distinguishing focal pock lesions were detected on CAM. Concerning electron microscopy, FPV appeared as enveloped quadrangular brick shaped Avipoxvirions. The neutralizing antibodies level against FPV were detected in all eight samples. Serum neutralization test showed a neutralization index of ≥ 1.6 in all serum samples, meanwhile ELISA test displayed an S/P ratio of ≥ 1.5 in the affected chickens. Notably, two positive FPV samples were sequenced then submitted to the GenBank (Sharquia-1 and Ismilia-2 with accession numbers; OR920788-OR920789). The phylogenetic tree construction based on the fpv167- (P4b) gene of FPV revealed high nucleotide identity with Elsharqyia_FWPV1, Elsharqyia_FWPV2 and Fowlpox-AN5, FWPVN, FWPVD (Egyptian isolates) with nucleotide identity percentage 100%, 99%, 100%, 99%,99%; respectively. Likewise, FPV isolates were of low homology with VSVRI-Vac (vaccinal strain) with 88% similarity. In context, the local recent our strains can be applied in vaccine production for appropriate vaccination programs in Egypt.

The lung is a complex organ, and its physiology and immunology are regulated by various immune molecules and cells. Lung surfactant, a mixture of phospholipids and proteins produced by the bronchiolar and type II alveolar epithelial cells, is one such important player in lung physiology. Compared to knowledge about the biology of the surfactant in rodents and humans, only limited data are available on the surfactant in the horse. Although there are data linking levels of surfactant proteins with respiratory disease in the horse, there are no data on the cellular localization of surfactant protein A (SP-A) and surfactant protein D (SP-D). A member of the tetraspanin family of proteins, CD9 is a cell-signaling and adhesion protein and its expression has been detected in both normal and cancer cells, including those in the lung. Because there are no immunolocalization data on SP-A, SP-D, and CD9 in the normal lungs of the horse, our objective was to conduct a light and electron microscopic immunocytochemical study on normal lungs of the horse. The data showed SP-A and SP-D in bronchiolar epithelial and type II alveolar epithelial cells. These proteins were also localized in type I alveolar epithelial cells, pulmonary intravascular macrophages, and neutrophils, which is likely an outcome of endocytosis of the proteins by these cells. CD9 was present in the airway and vascular smooth muscle cells, endothelium, and blood cells, but not in the airway epithelium. These new data provide a baseline to further examine the expression and functions of SP-A, SP-D, and CD9 proteins in inflammation associated with respiratory diseases in the horse.

Retinoids play an important role in lung development and immune response. The effects of retinoids are mediated through 2 families of retinoid receptors: retinoic acid receptors (RARs) and retinoid X receptors (RXRs), with alpha (α), beta (β), and gamma (γ) subtypes in each family. To date, no data exist on the expression pattern of retinoid receptors in lungs of cattle, dogs, and pigs. Because of the biomedical importance of retinoid receptors in inflammation and immune responses, Western blot, immunohistology, and immunoelectron microscopy were used to determine the expression of retinoid receptors in normal lungs of cattle, dogs, and pigs (n = 2 for each species). Western blot showed expression of all 6 retinoid receptor subtypes in pig lungs. Immunohistology data indicated differential expression of retinoid receptors in airway epithelium, vascular endothelium, alveolar/septal macrophages, and alveolar septum in all 3 species. Electron microscopy showed nuclear localization of retinoid receptors in neutrophils and pulmonary intravascular macrophages. Retinoic acid receptors (RAR) α subtype were localized in cytoplasmic vacuoles of pig monocytes. These data indicate constitutive expression of retinoid receptors in the lungs of cattle, dogs, and pigs.

The purpose of the study was to compare the conductance and mannitol permeability of canine colonic mucosa in response to carprofen or 2,4-dinitrophenol (DNP) with or without tempol pretreatment. Ten colonic mucosa sections per dog were mounted in Ussing chambers. Treatments were done in duplicate. Mucosa was exposed to carprofen (200 μg/mL) or DNP (0.25 mM), both with and without tempol (1 mM) pretreatment. Conductance was calculated every 15 min for 240 min. Mannitol flux was calculated over 3 consecutive 60-minute periods. Histology or electron microscopy was done after exposure. Conductance over time, mannitol flux, frequency of histologic categories, and electron microscopic changes were analyzed for treatment effects. The mean ± standard deviation (SD) conductance over time for carprofen or DNP-treated colons was not significantly different from control regardless of tempol pretreatment. Period 3 mannitol fluxes for carprofen and DNP-treated colon were not significantly different, but were greater than control. Period 3 mannitol flux for tempol + carprofen was significantly less than tempol + DNP-treated colon. Sloughing of cells and erosions were seen in the mucosa of carprofen-treated colon. Mitochondrial damage was seen more often in carprofen-treated than DNP-treated or control colon. Tempol pretreatment resulted in more ruptured mitochondria in the carprofen-treated colon; however, other mitochondrial changes were not significantly affected by tempol pretreatment in either carprofen or DNP treated colon. Treatment with carprofen or DNP increased the mannitol flux, but pretreatment with tempol mitigated the carprofen effect. It is apparent that structural mitochondrial damage occurs in the canine colonic mucosa after carprofen and DNP exposure.

Objective—To isolate and characterize bacteriophages with strong in vitro lytic activity against various pathogenic Pseudomonas aeruginosa strains isolated from dogs with ocular infections. Sample—26 genetically distinct P aeruginosa isolates. Procedures—P aeruginosa strains were derived from dogs with naturally acquired ulcerative keratitis. From a large-scale screening for bacteriophages with potential therapeutic benefit against canine ocular infections, 2 bacteriophages (P2S2 and P5U5) were selected; host ranges were determined, and phage nucleic acid type and genetic profile were identified via enzymatic digestion. Electron microscopy was used to characterize bacteriophage ultrastructure. Bacteriophage temperature and pH stabilities were assessed by use of double-layer agar overlay titration. A cocultivation assay was used to evaluate the effect of the bacteriophages on bacterial host growth. Results—P5U5 was active against all P aeruginosa isolates, whereas P2S2 formed lytic plaques on plates of 21 (80.8%) isolates. For each bacteriophage, the genomic nucleic acid was DNA; each was genetically distinct. Ultrastructurally, P2S2 and P5U5 appeared likely to belong to the Podoviridae and Siphoviridae families, respectively. The bacteriophages were stable within a pH range of 4 to 12; however, titers of both bacteriophages decreased following heating for 10 to 50 minutes at 45° or 60°C. Growth of each P aeruginosa isolate was significantly inhibited in coculture with P2S2 or P5U5; the dose response was related to the plaque-forming unit-to-CFU ratios. Conclusions and Clinical Relevance—Bacteriophages P2S2 and P5U5 appear to be good candidates for phage treatment of infection caused by pathogenic P aeruginosa in dogs.

This research illustrates the development of a new sliver nanoparticle (Ag-NPs) formulation. Its shape, size, solubility, and stability were characterized using Scanning Electron Microscope (SEM 3D), Transmission Electron Microscope (TEM 2D), Atomic Force Microscope (AFM), and Zeta size and Zeta potential. Exposure of Trichinella spiralis adult worms to 3, 6, 9 and 12 ppm of Ag-NPs each for 3,6,12 and 24 h. In vitro revealed a direct relation between mortalities and the tested drug concentration and exposure time. Anti- T. spiralis effect of Ag-NPs was evaluated by assessing mortality rate and damage in DNA by comet assay and by SEM analysis. Mean mortalities increased from 6.66% after exposure to 3.0 ppm/1 h to 100% after exposure to 12.0 ppm/12 h. The calculated LC50 was 3.0 ppm/10 h, 6 ppm/6 h, 9.0 ppm/4 h and 12.0 ppm/ 3.30 h, while LC100 was 9.0 ppm/24 h and 12.0 ppm/12 h. DNA genotoxic damage of dead worms was directly related to Ag-NPs concentrations for 12h using comet assay as expressed by variations in the percentage of DNA in the tail segment, tail length (μm), tail moment (μm), and olive tail moment. No significant difference (p ≤ .05) between the recorded mortalities and DNA damage between that obtained using the Ag-NPs LC100 and that recorded using Albendazole (50 mg/kg B.W.) for 12 h. SEM images on dead worms revealed clear morphological alteration, multiple vesicles, and blebs, detachment of the epidermis and the sub-epidermal layer with partial sloughing of the cuticle, and loss of normal creases, ridges, and annulations. These morphological alterations were directly related to the concentration of the tested Ag-NPs. The tested new formulation of Ag-NPs appears to be effective in the control of Trichinellosis as an alternative to other resistant drugs.