Genetic relatedness and diversity of Capillaria species infecting bayad (Bagrus bajad) in upper Egypt.

BACKGROUND: This study investigates the genetic characteristics of Capillaria isolates from the infected fish, Bagrus bajad, and their relation to human Capillaria philippinensis using Random Amplified Polymorphic DNA (RAPD-PCR) analysis. Fifteen fish Capillaria were isolated and compared to identified human C. philippinensis using six primers: M-are, M-1, G-7, G-11, G-15, and G-18. RESULTS: All six primers successfully amplified DNA, highlighting their efficacy in distinguishing between human and fish Capillaria isolates. The analysis revealed distinctive banding patterns between fish and human isolates, with variations in size and number of DNA fragments. Additionally, genetic similarity analysis showed intriguing patterns of relatedness, with certain pairs exhibiting high similarity percentages. Comparative assessment of RAPD polymorphism demonstrated consistent findings of 100% polymorphism across all primers. The Unweighted Pair Group Method with Arithmetic Mean Algorithm (UPGMA) evaluated the closest relationship between human and fish isolates. These results underscore the utility of RAPD analysis in delineating the genetic diversity among Capillaria isolates from different hosts. CONCLUSION: Overall, this study contributes to our understanding of the genetic variability and relatedness among Capillaria isolates, shedding light on their evolutionary dynamics and zoonotic potential.

Comparison of the diagnostic performance of microscopic examination, Copro-ELISA, and Copro-PCR in the diagnosis of Capillaria philippinensis infections.

Intestinal capillariasis is a parasitic zoonosis caused by the tiny nematode parasite Capillaria philippinensis. It is a major health problem that may lead to death if not diagnosed and treated appropriately. The difficulties in the diagnosis of C. philippinensis highlight the importance of developing accurate, sensitive, and specific methods for early diagnosis. This study aimed to detect the presence of C. philippinensis infection among 42 clinically suspected patients with certain criteria that are highly suggestive of capillariasis and to compare the diagnostic yield of microscopy, copro-ELISA, and PCR for the detection of copro-DNA. Sociodemographic characteristics and clinical data were also described for the infected group. Out of 42 patients, 10 were microscopically positive, 40 samples were positive by copro-ELISA, nested PCR detected 35 positive cases, with total detection rates of 23.8%, 95.2%, and 83.3% using direct microscopic examination, copro-ELISA, and PCR, respectively. The majority of positive cases were females, middle-aged people, and people from rural areas. The real number of cases infected with C. philippinensis may far exceed those estimated using microscopy. The diagnosis by copro-ELISA for the detection of C. philippinensis coproantigen and by nested PCR to identify parasite DNA revealed a higher number of positive cases. Using ELISA for the detection of coproantigen is a sensitive test that identifies the infection, yet it is not specific. Copro-DNA offers a satisfactory sensitive and specific method for the detection of infection in clinically suspected patients. The most susceptible individuals to C. philippinensis infection are females, middle-aged people, and people of low social standards. Intestinal capillariasis needs to be considered in patients who present with symptoms of chronic diarrhea and hypoalbuminemia because if these cases are left undiagnosed and untreated, they may suffer from lethal complications.

M-RNA Gene Expression of INF-Γ and IL-10 during Intestinal Phase of Trichinella spiralis after Myrrh and Albendazole Treatment.

BACKGROUND: The protective response developed against Trichinella spiralis infection provokes immune and inflammatory responses mediated by cytokines released from T helper cells. We aimed to evaluate the effect of albendazole or myrrh on the expression of IFN-γ and IL-10 in BALB/c mice infected with T. spiralis. METHODS: This study was done at the Animal House of Faculty of Medicine, Assiut University (Assiut, Egypt) from April to December 2015. Mice were infected with 300 T. spiralis larvae and treated with albendazole (50 mg/kg per day) or myrrh (500 mg/kg per day) for 3 consecutive days post-infection (pi). The expression of INF-γ and IL-10 was detected in the intestinal tissue by reverse transcription (RT) PCR. RESULTS: The expression of IFN-γ in mice treated with albendazole and myrrh was detected on days 3 and 15 pi respectively. In the control group, it was found on days 5, 10, 15 and 20 pi with the highest expression on day 15 pi. The expression of IL-10 was detected on days 3, 20 pi in the albendazole and myrrh treated groups, respectively. In the control group, IL-10 expression appeared on days 5 and day 20 pi. CONCLUSION: The target of albendazole and myrrh on the profile of IFN-γ and IL-10 on these cytokines were encouraging to reinforce their therapeutic use against trichinellosis.

Chemical composition and efficacy of some selected plant oils against Pediculus humanus capitis in vitro.

Natural compounds have been suggested as alternative sources for pediculosis capitis control. We aimed to investigate the chemical composition and evaluate the pediculicidal activity of spearmint, clove, cassia, thyme, eucalyptus, and anise essential oils in addition to sesame oil against human head lice in vitro. A filter paper contact bioassay method was used by applying 0.25 and 0.5 mg/cm(2) of each tested oil to filter paper in Petri dishes with 15 females head lice and another with ten nits. The lice mortalities were reported every 5 min for 180 min. The percentage of inhibition of hatch (PIH) was used to calculate ovicidal activity by daily microscopic inspections 5 days after the hatching of controls. Comparison with the widely used pediculicide (malathion) was performed. The most effective essential oil was spearmint followed by cassia and clove with KT50 values of 4.06, 7.62, and 12.12 at 0.5 mg/cm(2) and 8.84, 11.38, and 19.73 at 0.25 mg/cm(2), respectively. Thyme, eucalyptus, and anise were also effective adulticides with KT50 values of 18.61, 32.65, and 37.34 at 0.5 mg/cm(2) and 29.92, 43.16, and 45.37 at 0.25 mg/cm(2), respectively. Essential oils were also successful in inhibiting nymph emergence. Spearmint oil was the most effective, with a complete inhibition of emergence at 0.5 mg/cm(2). Sesame fixed oil did not show any adulticidal or ovicidal activity against head lice in vitro. The observed insecticidal activity was comparable to malathion. The results herein described the effectiveness of these essential oils as potential pediculicides for head lice control. Incorporation of essential oils in pediculicide formulations needs proper formulation and clinical trials.

Resistance and Susceptibility to Malarial Infection: A Host Defense Strategy against Malaria.

BACKGROUND: In an effort to understand what limits the virulence of malaria parasites in relation to the host genetic and immunogenic background, we investigated the possibility that the parasite and host genotype crossover interactions constrain virulence. METHODS: Two groups of mice from different genotypes were used (C57BL/6 (B6) and DBA/2 mice). The mice were infected with a virulent parasite line Plasmodium yoelii 17XL (P. yoelii 17XL). Parasitemia, hematocrit value and lymphocytes yielded by livers and spleens were evaluated. Fluorescence Activated Cell Sorting (FACS) analysis illustrated phenotypic characterization of lymphocytes. RESULTS: Infection with P. yoelii 17XL did not result in the death of DBA/2 mice. In contrast, B6 mice developed significantly high parasitemia and succumbed to death. Using (FACS) analysis, DBA/2 mice were found to experience a marked expansion of interleukin (IL)-2Rß(+) CD3(int) cells and γδ T cells in the liver, especially in the recovery phase. The expansion of unconventional T cells (i.e. B220(+) T cells) was also marked in DBA/2 mice. CONCLUSION: The outcome of murine malaria infections depends on the dynamic interplay between the immune-mediator and the genotype of the host.

Capillaria philippinensis in Upper Egypt: has it become endemic?

The goal of this study was to present an overview of human infections with Capillaria philippinensis, a new emerging parasite in Upper Egypt. The study included 21 inpatients who had been admitted to the Assiut University Hospital. Patients suffered from intermittent abdominal pain, borborygmi, chronic diarrhea lasting for several weeks, and marked weight loss. Hypoalbuminemia and low serum levels of potassium, calcium, and sodium were detected in most patients. A stool examination was performed using direct smears and the formalin-ether concentration method. Intact adult worms and/or eggs were evaluated using a light microscope and processed for scanning electron microscopy. The examination by light microscopy illustrated the general morphology of different stages. Using scanning electron microscopy, intestinal villi were found partially covering the cuticle of the adult worms, which provided evidence for the invasion of adult worms into the jejunal mucosa. Two distinct types of eggs, thick-shelled and thin-shelled, were identified and measured.

Cytokine profile of murine malaria: stage-related production of inflammatory and anti-inflammatory cytokines.

Balance between inflammatory and anti-inflammatory cytokines may be important in malaria presentation and outcome. To clarify cytokine interactions that produce pathology of malaria and control infection, C57BL/6 mice were infected with 10(4) parasitized RBCs from a non-lethal strain of Plasmodium yoelii. Kinetics was monitored showing the course of parasitemia, and cytokines were determined by RT-PCR from liver and spleen tissues. Inflammatory cytokines such as interferon-γ (IFNγ), interleukin (IL)-12, IL-6, tumor necrosis factor-α (TNFα) and anti-inflammatory cytokines, including IL-4 and IL-10, were investigated as key molecules that interact with immune cells in the activation of the immune responses. The production of IFNγ mRNA was found to be higher on day 7 than on day 21 after infection, and IL-12 and IL-6 showed higher expression in the liver than in the spleen. Though TNFα was highly expressed on day 14 after infection and on day 21 in the liver, such expression was decreased on day 21 in the spleen. Anti-inflammatory cytokines showed high expression in both the liver and spleen. The results suggest that a relative balance between inflammatory and anti-inflammatory cytokines is crucial and that the increase of inflammatory cytokine levels during the acute phase of malaria may reflect an early and effective immune response.The counteraction effect of anti-inflammatory cytokines is thought to play a role in limiting progression from uncomplicated malaria to severe life-threatening complications.

Comparative study between non lethal and lethal strains of Plasmodium yoelii with reference to its immunological aspect.

Innate immunity has an important role in the protection against malaria. To clarify the effect on non lethal and lethal strain of Plasmodium yoelii, comparison between two groups of C57BL/6 mice infected with 10(4) parasitized RBCs was performed. Liver and spleen mononuclear cells were isolated and analyzed by flow cytometry. The parasite appeared in blood on day 3 in both strains, with non lethal infection parasitemia reached a peak of 60% on day 14 and mice completely recovered, while in lethal infection parasitemia was 80% on day 7 and mice succumbed to death. In non lethal strain, mice became anemic and the hematocrit percentage returned to its normal value during recovery, while in the lethal strain mice were severely anemic before death. The major expanding cells were found to be TCR Intermediate (TCRint) cells, mainly NK1.1 subset, these TCRint cells were distinguished from conventional T cells of thymic origin. CD4 & CD8 cells increased in both strains. During malarial infection, the population of conventional T cells did not increase and usually associated with thymic atrophy. The present results showed that TCRint cells were intimately associated with the protection against malarial infection in both non lethal and lethal strains but the mice died in lethal infection due to the massive destruction of red blood cells leading to fatal anemia.

Modulation of the endocrine and immune systems by well-controlled hyperthermia equipment.

Since high levels of hyperthermia induce immunosuppression to a certain extent (i.e., granulocytosis and lymphocytopenia) in patients, we applied mild hyperthermia in volunteers using equipment enabling well-controlled hyperthermia. Restricted control of rectal temperature at 39.4 (+/- 0.2) degrees C for 30 min was conducted and various parameters of the body were examined. The most prominent change observed during exposure to hyperthermia was elevated levels of pH and PO(2) in the blood, even in the venous blood. A transient elevation of ACTH, cortisol and growth hormone in the blood was also seen during this time. In parallel with this phenomenon, the number of total lymphocytes and those of its subsets (especially CD57(+) or CD56(+) NK cells and NKT cells) increased. More interestingly, the proportion of HLA-DR (MHC class II antigens) increased in NK and NKT cells, and their intensity on the surface of CD20(+) B cells increased. These results suggest that mild hyperthermia is important for modulation of the functions of the circulatory, endocrine and immune systems.

Protection against malaria due to innate immunity enhanced by low-protein diet.

Mice were fed ad libitum with a normal diet (25% protein) or low-protein diets (0-12.5% protein) for a wk and then infected with a nonlethal or lethal strain of Plasmodium yoelii, that is, blood stage infection. The same diet was continued until recovery. Mice fed with a normal diet showed severe parasitemia during nonlethal infection, but survived the infection. They died within 2 wk in the case of lethal infection. However, all mice fed with low-protein diets survived without apparent parasitemia (there were small peaks of parasitemia) in cases of both nonlethal and lethal strains. These surviving mice were found to have acquired potent innate immunity, showing the expansion of NK1.1 -TCRint cells and the production of autoantibodies during malarial infection. Severe combined immunodeficiency (scid) mice, which lack TCRint cells as well as TCRhigh cells, did not survive after malarial infection of lethal strain of P. yoelii, even when low-protein diets were given. These results suggest that low-protein diets enhanced innate immunity and inversely decreased conventional immunity, and that these immunological deviations rendered mice resistant against malaria. The present outcome also reminds us of our experience in the field study of malaria, in which some inhabitants eventually avoided contracting malaria even after apparent malarial infection.

Reasons why DBA/2 mice are resistant to malarial infection: expansion of CD3int B220+ gammadelta T cells with double-negative CD4- CD8- phenotype in the liver.

DBA/2 (H-2(d)) mice are known to be more resistant than C57BL/6 (B6, H-2(b)) mice to the non-lethal 17XNL strain of Plasmodium yoelii. This is a very strange phenomenon because the functions of conventional T cells, especially CD8(+) T cells, are known to be somewhat lower in DBA/2 mice than in other strains of mice. We examined herein how immune responses differed between DBA/2 mice and B6 mice during malarial infection. DBA/2 mice and (DBA/2 x B6)F(1) (BDF(1), H-2(b/d)) mice were found to have milder parasitaemia and to recover more quickly from malarial infection than B6 mice. These DBA/2 and BDF(1) mice were also found to experience a marked expansion of interleukin (IL)-2Rbeta(+) CD3(int) cells and gammadelta T cells in the liver, especially in the recovery phase. The expansion of unconventional T cells (i.e. B220(+) T cells) was also marked in DBA/2 and BDF(1) mice. The majority of B220(+) T cells were gammadelta T cells and these T cells were double-negative CD4(-) CD8(-). More importantly, the production of immunoglobulin M (IgM)-type anti-DNA autoantibody was also higher in DBA/2 and BDF(1) mice than in B6 mice. In conjunction with data on cytokine production, these results indicate that primitive T and B cells, namely autoreactive extrathymic T cells and autoantibody-producing B cells, may be much more activated in DBA/2 mice and therefore resistant to the non-lethal 17XNL strain of P. yoelii.

Essential role of extrathymic T cells in protection against malaria.

Athymic nude mice carry neither conventional T cells nor NKT cells of thymic origin. However, NK1.1(-)TCR(int) cells are present in the liver and other immune organs of athymic mice, because these lymphocyte subsets are truly of extrathymic origin. In this study, we examined whether extrathymic T cells had the capability to protect mice from malarial infection. Although B6-nu/nu mice were more sensitive to malaria than control B6 mice, these athymic mice were able to survive malaria when a reduced number of parasitized erythrocytes (5 x 10(3) per mouse) were injected. At the fulminant stage, lymphocytosis occurred in the liver and the major expanding lymphocytes were NK1.1(-)TCR(int) cells (IL-2Rbeta(+)TCRalphabeta(+)). Unconventional CD8(+) NKT cells (V(alpha)14(-)) also appeared. Similar to the case of B6 mice, autoantibodies (IgM type) against denatured DNA appeared during malarial infection. Immune lymphocytes isolated from the liver of athymic mice which had recovered from malaria were capable of protecting irradiated euthymic and athymic mice from malaria when cell transfer experiments were conducted. In conjunction with the previous results in euthymic mice, the present results in athymic mice suggest that the major lymphocyte subsets associated with protection against malaria might be extrathymic T cells.