Molecular Survey of Anaplasma capra in Goats in Van Province, Eastern Türkiye.

BACKGROUND: A newly discovered zoonotic infection carried by ixodid ticks, Anaplasma capra, affects a wide variety of hosts, including numerous mammals. A. capra most likely infects erythrocytes or endothelial cells in mammals. This study aimed to investigate the A. capra pathogen in goats in Türkiye's Van province. METHODS: A total of 200 goat blood samples were examined. Goat samples were subjected to partial amplification of the gltA gene fragment using a nested polymerase chain reaction. RESULTS: A. capra DNA was detected in 0.5% of goat blood samples. Phylogenetic analysis of a partial gltA gene fragment showed that the Eastern Türkiye isolate, closely grouped with A. capra isolates reported from wild and domestic ruminants in France, Türkiye, and Kyrgyzstan, formed a distinct clade. CONCLUSIONS: This is the first report of A. capra in goats in Van province, Eastern Türkiye.

Co-Immunization with DNA Vaccines Expressing SABP1 and SAG1 Proteins Effectively Enhanced Mice Resistance to Toxoplasma gondii Acute Infection.

Toxoplasma gondii (T. gondii) has many intermediate hosts, obligately invades nucleated cells, and seriously threatens human and animal health due to a lack of effective drugs and vaccines. Sialic acid-binding protein 1 (SABP1) is a novel invasion-related protein that, like surface antigen 1 (SAG1), is found on the plasma membrane of T. gondii. To investigate the immunogenicity and protective efficacy of DNA vaccines expressing SABP1 and SAG1 proteins against T. gondii acute infection, the recombinant plasmids pVAX1-SABP1 and pVAX1-SAG1 were produced and administered intramuscularly in Balb/c mice. Serum antibody levels and subtypes, lymphocyte proliferation, and cytokines were used to assess immunized mice's humoral and cellular immune responses. Furthermore, the ability of DNA vaccines to protect mice against T. gondii RH tachyzoites was tested. Immunized mice exhibited substantially higher IgG levels, with IgG2a titers higher than IgG1. When the immune group mice's splenocytes were stimulated with T. gondii lysate antigen, Th1-type cytokines (IL-12p70, IFN-γ, and IL-2) and Th2-type cytokine (IL-4) increased significantly. The combined DNA vaccine significantly increased the immunized mouse survival compared to the control group, with an average death time extended by 4.33 ± 0.6 days (p < 0.0001). These findings show that DNA vaccines based on the SABP1 and SAG1 genes induced robust humoral and cellular immunity in mice, effectively protecting against acute toxoplasmosis and potentially serving as a viable option for vaccination to prevent T. gondii infection.

Anti-proliferative and apoptosis induction activities of curcumin on Leishmania major.

Leishmaniasis is a major vector-borne disease triggered by an obligate intracellular protozoan parasite of the genus Leishmania and transmitted by the bite of phlebotomine female sand flies. This parasite causes a wide range of human diseases, from localized self-healing cutaneous lesions to fatal visceral infections. The aim of this study was to investigate the cytotoxic, antiproliferative, and apoptotic effects of curcumin on Leishmania major promastigotes (MHOM/SA/84/JISH) and to assess these effects on the cell cycle of promastigotes. The MTT colorimetric assay was used to evaluate the cytotoxicity and proliferation of promastigotes. Additionally, flow cytometry was used to analyze the cell cycle. The Annexin V/propidium iodide staining technique followed by flow cytometry was used to study the cell death induced by curcumin. In this study curcumin showed a potent antileishmanial effect, exhibiting cytotoxicity against L. major promastigotes. At 80µM, the survival in curcumin treated promastigotes reached 22%; however, the median lethal concentration of curcumin (LC50) was 35µM. The drug exerted its cytotoxic effect by inducing apoptosis. Curcumin-induced cell death in promastigotes reached 82.5% at 80µM concentration. In addition, curcumin delayed the cell cycle in the S-phase inhibiting cell proliferation. Thus, curcumin was shown to be effective against L. major promastigotes. Therefore, curcumin merits further research studies to demonstrate its efficacy in treating cutaneous leishmaniasis.

Morphological, Molecular, and Pathological Appraisal of Hymenolepis nana (Hymenolepididae) Infecting Laboratory Mice (Mus musculus).

Hymenolepis nana, typically a parasite found in conventionally established mouse colonies, has zoonotic potential characterized by autoinfection and direct life cycle. The objective of this study was to determine the rate of parasite infection in laboratory mice. The hymenolepidide cestode infected 40% of the 50 mice sampled. The rate of infection in males (52%) was higher than in females (28%). Morphological studies on the cestode parasite showed that worms had a globular scolex with four suckers, a retractable rostellum with 20-30 hooks, and a short unsegmented neck. In addition, the remaining strobila consisted of immature, mature, and gravid proglottids, irregularly alternating genital pores, lobulated ovaries, postovarian vitelline glands, and uteri with up to 200 eggs in their gravid proglottids. The parasite taxonomy was confirmed by using molecular characterization based on the sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (mtCOX1) gene. The parasite recovered was up to 80% identical to other species in GenBank. High blast scores and low divergence were noted between the isolated parasite and previously described H. nana (gb| AP017666.1). The phylogenetic analysis using the COX1 sequence places this hymenolepidid species of the order Cyclophyllidea.

Henneguya collaris sp. nov., (Myxosporidia), parasite of the Greenband Parrotfish Scarus collana Rüppell, 1835 (Actinopterygii, Scaridae) from the Red Sea, Egypt. A light and electron microscopic study.

The phylum Myxozoa comprises more than 2180 species, almost all of which are considered to be obligate parasites of aquatic fishes. In the present study, Henneguya collaris sp. nov. is the first described histozoic myxozoan species of the genus Henneguya infecting the kidney of the greenband parrotfish Scarus collana (Actinopterygii, Scaridae). One hundred and eighty specimens of fish were collected randomly during the period from September 2014 to October 2015 from boat landing sites and the market places at Hurghada City along the Red Sea in Egypt. Of these, 90 (50 %) specimens were infected. Light microscopic examination showed that the infection was detected as mature spores with two polar capsules regularly arranged at the anterior pole of each spore and extruded polar filaments free in the kidney tissue. The spore body was oval in shape, measured 7.1 ± 0.2 (6.2-8.4) µm in length and 6.3 ± 0.2 (5.8-7.0) µm in width, with a bifurcated caudal process of equal length, reaching 6.3 ± 0.2 (5.8-7.0) µm in length. Polar capsules were 3.4 ± 0.2 (3.0-4.2) µm in length and 1.9 ± 0.2 (1.6-2.4) µm in width with 6-8 (10) turns of polar filaments. Ultrastructural analysis showed that the spore development was asynchronous. Sporogenesis, capsulogensis, valvogenesis, and spore maturation of the present parasite were also described. The present species was compared morphologically with the spore characteristics of the most similar species of Henneguya spp. recorded previously from different geographical areas taking into account the stage and dimensions of the spore body, tails, and polar filament coils, including their number and the most important characteristic features that distinguish them from the present species. Considering the data obtained, the material described herein represents a new species and the name Henneguya collaris sp. nov. is proposed.

Testicular myxosporidiasis and ultrastructural characteristics of Myxobolus bufonis (Myxobolidae) infecting the Egyptian toad Bufo regularis (Bufonidae). A light and electron microscopic study.

The phylum Myxozoa comprises more than 2180 species, almost all of which are considered to be obligate parasites of aquatic fishes and amphibians. They are dangerous pathogens responsible for severe economic losses. From March to September 2014, 40 adult male Bufo regularis (Bufonidae) captured from different areas at Giza province, Egypt, were surveyed for myxosporean parasitic infection. Of these, 22 (55%) were infected by histozoic plasmodia, which produced spores after rupture belonging to Myxosporidia. The present investigation introduced a new data for the recorded parasite observed by light and transmission electron microscopy. The infection was diagnosed as large clusters of macroscopic plasmodia embedded in the testicular tissue causing distortion at the site of infection. The host reaction was manifested by the encapsulation of the plasmodia with a thick layer of connective tissue. Plasmodia were whitish in color, elliptical to ovoid in shape measuring 0.54 ± 0.2 (0.34-0.63) mm in diameter. The spores were subspherical, reaching 7.1 ± 0.2 (6.2-8.4) µm in length and 6.3 ± 0.2 (5.8-7.0) µm in width with two equal-sized polar capsules regularly arranged at the anterior pole of each spore. They were 3.4 ± 0.2 (3.0-4.2) µm in length and 1.9 ± 0.2 (1.6-2.4) in width with 6-8 turns of polar filaments. Ultrastructural analysis showed that the plasmodia were surrounded by a plasma membrane with numerous projections and pinocytotic channels extended toward the host cell. The generative cells and the different developmental stages were arranged at the periphery of the plasmodia while immature and mature spores were centrally arranged. Sporogenesis, capsulogenesis, valvogenesis, and spore maturation of the present parasite were also described.

Molecular cloning, characterization and predicted structure of a putative copper-zinc SOD from the camel, Camelus dromedarius.

Superoxide dismutase (SOD) is the first line of defense against oxidative stress induced by endogenous and/or exogenous factors and thus helps in maintaining the cellular integrity. Its activity is related to many diseases; so, it is of importance to study the structure and expression of SOD gene in an animal naturally exposed most of its life to the direct sunlight as a cause of oxidative stress. Arabian camel (one humped camel, Camelus dromedarius) is adapted to the widely varying desert climatic conditions that extremely changes during daily life in the Arabian Gulf. Studying the cSOD1 in C. dromedarius could help understand the impact of exposure to direct sunlight and desert life on the health status of such mammal. The full coding region of a putative CuZnSOD gene of C. dromedarius (cSOD1) was amplified by reverse transcription PCR and cloned for the first time (gene bank accession number for nucleotides and amino acids are JF758876 and AEF32527, respectively). The cDNA sequencing revealed an open reading frame of 459 nucleotides encoding a protein of 153 amino acids which is equal to the coding region of SOD1 gene and protein from many organisms. The calculated molecular weight and isoelectric point of cSOD1 was 15.7 kDa and 6.2, respectively. The level of expression of cSOD1 in different camel tissues (liver, kidney, spleen, lung and testis) was examined using Real Time-PCR. The highest level of cSOD1 transcript was found in the camel liver (represented as 100%) followed by testis (45%), kidney (13%), lung (11%) and spleen (10%), using 18S ribosomal subunit as endogenous control. The deduced amino acid sequence exhibited high similarity with Cebus apella (90%), Sus scrofa (88%), Cavia porcellus (88%), Mus musculus (88%), Macaca mulatta (87%), Pan troglodytes (87%), Homo sapiens (87%), Canis familiaris (86%), Bos taurus (86%), Pongo abelii (85%) and Equus caballus (82%). Phylogenetic analysis revealed that cSOD1 is grouped together with S. scrofa. The predicted 3D structure of cSOD1 showed high similarity with the human and bovine CuZnSOD homologues. The Root-mean-square deviation (rmsd) between cSOD1/hSOD1 and cSOD1/bSOD1 superimposed structure pairs were 0.557 and 0.425 A. The Q-score of cSOD1-hSOD1 and cSOD1-bSOD1 were 0.948 and 0.961, respectively.

Ultrastructure and host parasite relationships of Kudoa pagrusi (Myxozoa) infecting the heart muscles of sea bream Pagrus pagrus (L.) from the Red Sea.

The present study is a part of a continuous investigation of myxosporean parasites-infecting fish of the Red Sea using light and electron microscopy. Out of 120, 80 (67%) Pagrus pagrus fish were found to be naturally infected with Kudoa pagrusi. The infection was intensive and appeared as clusters of ovoid to ellipsoidal plasmodia being restricted to the cardiac muscles. Histological studies elaborated tissue distortion at the sites of infection and the adjacent layers. The development of the plasmodia reduced the functional area of the heart muscle. Ultrastructural analysis showed that the plasmodia were surrounded by single-unit membrane with numerous projections and pinocytotic channels extended toward the host cell. The generative cells and the different developmental stages were arranged at the periphery of the plasmodia while immature and mature spores were centrally arranged. The present study showed the main criteria of this genus: the spores possess four polar capsules with four shell valves.

Ultrastructure, development, and host-parasite relationship of a new species of the genus Pleistophora--a microsporidian parasite of the marine fish Epinephelus chlorostignei.

The life cycle of a new microsporidian of the genus Pleistophora is described. This parasite infects the epithelial cells of the gut and the peritoneal cavity of the Red Sea fish, Epinephelus chlorostignei. All stages develop within a special structure, the sporophorocyst, which is covered by a thick dense wall. This wall grows along with the growth of the parasites inside. Meronts are uni- to binucleate, which divide and constantly give rise to sporonts. During transition to sporonts, the cell border of the meronts increases its thickness, temporarily featuring thick irregular projections. Eventually, a uniform thick sporont wall is formed; then, the sporont cells detach themselves from the wall (future wall of the sporophorous vesicle, SPV) and start a series of divisions to produce sporoblasts. The SPV wall is compact, has no pores, and consists of two layers. Mature spores measure about 2.0 x 1.8 microm. They possess a polar filament with 20-28 coils, a posterior vacuole, and a polaroplast made up of an outer part of dense and closely spaced lamellae encircling an inner part of widely spaced lamellae. All morphological and ultrastructural features indicate that the described microsporidian parasite belongs to the genus Pleistophora.