Ameliorative effects of propolis and wheat germ oil on acute toxoplasmosis in experimentally infected mice are associated with reduction in parasite burden and restoration of histopathological changes in the brain, uterus, and kidney.

Toxoplasmosis continues to be a prevalent parasitic zoonosis with a global distribution. This disease is caused by an intracellular parasite known as Toxoplasma gondii, and the development of effective novel drug targets to combat it is imperative. There is limited information available on the potential advantages of wheat germ oil (WGO) and propolis, both individually and in combination, against the acute phase of toxoplasmosis. In this study, acute toxoplasmosis was induced in Swiss albino mice, followed by the treatment of infected animals with WGO and propolis, either separately or in combination. After 10 days of experimental infection and treatment, mice from all groups were sacrificed, and their brains, uteri, and kidneys were excised for histopathological assessment. Additionally, the average parasite load in the brain was determined through parasitological assessment, and quantification of the parasite was performed using Real-Time Polymerase Chain Reaction targeting gene amplification. Remarkably, the study found that treating infected animals with wheat germ oil and propolis significantly reduced the parasite load compared to the control group that was infected but not treated. Moreover, the group treated with a combination of wheat germ oil and propolis exhibited a markedly greater reduction in parasitic load compared to the other groups. Similarly, the combination treatment effectively restored the histopathological changes observed in the brain, uterus, and kidney, and the scoring of these reported lesions confirmed these findings. In summary, the present results reveal intriguing insights into the potential therapeutic benefits of wheat germ oil and propolis in the treatment of acute toxoplasmosis.

Wheat Germ Oil and Propolis Decrease Parasite Burden and Restore Marked Histopathological Changes in Liver and Lung in Mice with Chronic Toxoplasmosis.

Toxoplasmosis is a parasitic zoonotic disease with a worldwide distribution. Its effects can be critical in immunocompromised patients. However, there is a limited availability of effective, low-toxicity drugs against this disease, particularly in its chronic form. The present study evaluated the effect of propolis and wheat germ oil (WGO) as safe, natural products to reduce Toxoplasma cysts in experimentally infected mice. For the experiment, five groups (10 mice per group) were examined: Group 1: negative control (noninfected, nontreated); Group 2: positive control (infected, nontreated); Group 3: infected and treated with WGO at a dose of 0.2 mg/1.5 mL per kg body weight/day; Group 4: infected and treated with 0.1 mL propolis extract/day; and Group 5: infected and treated with a combination of WGO and propolis at the same doses as Group 3 and 4. After the mice were sacrificed, liver and lung specimens underwent histopathological examination, and the parasite burden was investigated by parasitological methods and quantified using real-time polymerase chain reaction. Notably, the results showed a substantial decrease in parasitic burden in Group 5 compared to the control group. These results were further confirmed by molecular analysis and quantification of the DNA concentration of the Toxoplasma P29 gene after treatment in all tested samples. Furthermore, the combination of propolis and WGO restored all histopathological changes in the liver and lungs. Taken together, these findings provide remarkably promising evidence of the effects of the combination of WGO and propolis against chronic toxoplasmosis in mice.

Novel insights on the potential activity of propolis and wheat germ oil against chronic toxoplasmosis in experimentally infected mice.

The use of apitherapy and natural herbal medicines for combating toxoplasmosis has garnered major attention from many researchers. However, there is no available information regarding the potential use of a combination of propolis and wheat germ oil (WGO) in the treatment of toxoplasmosis. In the present study, the potential effects of propolis, WGO, and their combination in the treatment of chronic toxoplasmosis in Swiss albino mice were investigated. Following induction of chronic toxoplasmosis, the potential antiparasitic effects of these substances were evaluated by parasitological assessment and by counting of Toxoplasma cysts. Additionally, the effects of the treatments on parasite loads were analyzed using TaqMan real-time quantitative PCR targeting the Toxoplasma P29 gene followed by investigation of the major histopathological changes in the brain, uterus, and kidney. Interestingly, the combination of propolis and WGO significantly (P ≤ 0.05) decreased the parasite burden in experimental animals compared with burdens seen in groups treated with propolis or WGO alone. Furthermore, the quantification of the DNA concentrations of Toxoplasma P29 gene after the treatment with propolis and WGO revealed a reduction in parasite load in treated groups versus the control group (infected untreated animals). Importantly, the severity of histopathological lesions was significantly (P ≤ 0.05) improved following treatment with propolis and WGO. Collectively, the present study indicated a potential novel role for propolis and WGO as an active apitherapy and natural herbal medication for treating chronic toxoplasmosis, combat the disease, and which could also help overcome the side effects of chemical drugs.

Effectiveness of new selenium-enriched mutated probiotics in reducing inflammatory effects of piroxicam medication in liver and kidney.

Piroxicam is used to treat the pain, swelling, and stiffness associated with osteoarthritis and rheumatoid arthritis, but it has many side effects, such as hypertension, elevation of liver enzymes, and hepatitis. This study used selenium-enriched probiotics to reduce the side effects of piroxicam on the liver and kidney tissues and functions. Forty-eight male albino mice were randomly assigned to control, piroxicam (P), piroxicam plus selenium-enriched Lactobacillus plantarum PSe40/60/1 (P + SP), piroxicam plus selenium-enriched Bifidobacterium longum BSe50/20/1 (P + SB), selenium-enriched L. plantarum PSe40/60/1 (SP), and selenium-enriched B. longum BSe50/20/1 (SB) groups. In this study, the function of the liver and kidney was biochemically determined; the histopathology of the liver and kidney tissues was microscopically examined and the expression of inflammatory and anti-inflammatory genes in liver and kidney tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Liver and kidney functions were significantly reduced in the piroxicam group compared with control. Liver and kidney tissues were damaged in the piroxicam group while they appeared more or less normal in the SB group. The expression of inflammatory genes was significantly up-regulated in the liver and kidney tissues of the piroxicam group compared to the control group. The expression of anti-inflammatory genes was significantly down-regulated in the liver and kidney of the piroxicam group and up-regulated in the liver and kidney of the SB group compared to the control group. Therefore, these mutated strains of probiotics were useful in reducing the side effects of the piroxicam drug on the liver and kidney.

Tamoxifen Increased Parasite Burden and Induced a Series of Histopathological and Immunohistochemical Changes During Chronic Toxoplasmosis in Experimentally Infected Mice.

The global distribution of breast cancer and the opportunistic nature of the parasite have resulted in many patients with breast cancer becoming infected with toxoplasmosis. However, very limited information is available about the potential effects of tamoxifen on chronic toxoplasmosis and its contribution to the reactivation of the latent infection. The present study investigated the potential effects of tamoxifen on chronic toxoplasmosis in animal models (Swiss albino mice). Following induction of chronic toxoplasmosis and treatment with the drug for 14 and 28 days, the anti-parasitic effects of tamoxifen were evaluated by parasitological assessment and counting of Toxoplasma cysts. In addition, the effects of the drug on the parasite load were evaluated and quantitated using TaqMan real-time quantitative PCR followed by investigation of the major histopathological changes and immunohistochemical findings. Interestingly, tamoxifen increased the parasite burden on animals treated with the drug during 14 and 28 days as compared with the control group. The quantification of the DNA concentrations of Toxoplasma P29 gene after the treatment with the drug revealed a higher parasite load in both treated groups vs. control groups. Furthermore, treatment with tamoxifen induced a series of histopathological and immunohistochemical changes in the kidney, liver, brain, and uterus, revealing the exacerbating effect of tamoxifen against chronic toxoplasmosis. These changes were represented by the presence of multiple T. gondii tissue cysts in the lumen of proximal convoluted tubules associated with complete necrosis in their lining epithelium of the kidney section. Meanwhile, liver tissue revealed multiple T. gondii tissue cysts in hepatic parenchyma which altered the structure of hepatocytes. Moreover, clusters of intracellular tachyzoites were observed in the lining epithelium of endometrium associated with severe endometrial necrosis and appeared as diffuse nuclear pyknosis combined with sever mononuclear cellular infiltration. Brain tissues experienced the presence of hemorrhages in pia mater and multiple T. gondii tissue cysts in brain tissue. The severity of the lesions was maximized by increasing the duration of treatment. Collectively, the study concluded novel findings in relation to the potential role of tamoxifen during chronic toxoplasmosis. These findings are very important for combating the disease, particularly in immunocompromised patients which could be life-threatening.

Study on prevalence and bacterial etiology of mastitis, and effects of subclinical mastitis and stage of lactation on SCC in dairy goats in Egypt.

In Egypt, inadequate information on prevalence and epidemiology of caprine mastitis is available. This study was designed to investigate prevalence and etiological agents of caprine mastitis and assess the efficacy of somatic cell count (SCC) as marker of subclinical mastitis (SCM) in dairy goats. This study was carried out on 249 randomly selected lactating goats in different lactation stages and examined clinically. Of these animals, 477 milk samples were aseptically collected and screened for bacterial carriage. SCC was assessed in 234 apparently normal milk samples, and SCC ≥ 106 cells/ml was indicator for SCM. Prevalence of clinical mastitis (CM) was 33.73% and 16.87% at animal and udder-half levels, respectively. SCM was 52.56% in the apparently healthy halves. Culture results proved single infection in 49.69% of samples, mixed infection in 23.9% of samples, and 26.41% of samples were negative. Coagulase negative staphylococci (CNS) were the most predominant bacteria (58.75%), then Staphylococcus aureus (S. aureus) (24.375%), and Streptococci (1.875%) were the least. No significant difference was recorded between mean of SCC in bacteriologically positive and negative samples, neither in those with SCC ≤ 106 nor with SCC ≥ 106 cells/ml both in middle and late lactation stages. Besides, the percentage of animals harboring SCC ≥ 106 cells/ml and negative for bacteriology in late lactation stage was 3 times (28.57%) more than in midlactation (9.3%). We can assume that SCC is not proper indicator for intra-mammary inflammation (IMI) in goats, and bacteriological examination remains more efficient, despites being time consuming and expensive.

Corynebacterium pseudotuberculosis mastitis in Egyptian dairy goats.

BACKGROUND AND AIM: Mastitis is an important threat facing goat milk industry and is the most common cause of culling. Efficient control of mastitis, based on efficient diagnosis of diseased animals, would improve milk production and reproductive efficiency. In subclinical mastitis (SCM), infected goats demonstrate neither udder symptoms nor abnormal milk. Corynebacterium pseudotuberculosis is an infectious causative agent of mastitis, mostly results as an extension of infection from the supramammary lymph node, and causes financial losses in the goat industry. This study aimed to estimate the prevalence of SCM with emphasis on C. pseudotuberculosis mastitis in Egyptian dairy goats in the selected farms. MATERIALS AND METHODS: A total of 336 half milk samples were collected from 177 dairy goats of various crossbreeds, in mid-to-late lactation period, after clinical examination. All samples were examined bacteriologically, while somatic cell count (SCC) was determined only in 180 half milk samples of the clinically healthy milk samples. The isolated and identified C. pseudotuberculosis was examined for evidence of virulence genes (Phospholipase D [pld] and ß-subunit of RNA polymerase [rpoB]) by polymerase chain reaction (PCR). RESULTS: The prevalence of clinical mastitis was 30.5%, while 69.5% of animals were apparently healthy and secreted milk was normal. Of those 180 clinically healthy half milk samples, 96 milk samples (53.33%) showed SCM as detected by SCC (SCC ≥1,000,000 cells/ml). Coagulase-negative staphylococci were the most prevalent bacteria (41.96%), then Staphylococcus aureus (37.5%) and C. pseudotuberculosis (7.14%). Molecular diagnosis of virulence genes revealed evidence of pld gene in 16 isolates (66.66%), and rpoB gene in 6 samples (25%) of the 24 bacteriologically isolated C. pseudotuberculosis. Here, we describe, for the 1st time, isolation and identification of C. pseudotuberculosis from milk of does suffering from SCM in Egypt. CONCLUSION: C. pseudotuberculosis must be considered for routine bacteriological examination of milk from dairy goats, particularly herds with a history of caseous lymphadenitis. Pld gene-based PCR is more reliable than rpoB gene-based ones for the diagnosis of C. pseudotuberculosis.

Seroprevalence, isolation, molecular detection and genetic diversity of Toxoplasma gondii from small ruminants in Egypt.

Toxoplasmosis is an infectious zoonotic disease caused by protozoan Toxoplasma gondii. Detection of T. gondii infection with touchy and particular strategies is a key advance to control and prevent toxoplasmosis. Genotyping can explain the virulence, epidemiology and setting up new methodologies for diagnosis and control in human and animals. The point of this study was to assess the seroprevalence of T. gondii in sheep and goat in Egypt and to comprehend the genetic variety of T. gondii isolates circling in Egypt. Blood samples were gathered from 113 ewes and 95 she-goats from three Egyptian governorates (Cairo, Giza and Al-Sharkia). Also blood and tissue samples were gathered from 193 sheep and 51 goats from Cairo and Giza abattoirs. All samples were assayed serologically utilizing ELISA and OnSite Toxo IgG/IgM Rapid test cassettes (OTRT) tests and the tissue samples of the seropositive animals were digested and microscopically examined then bio-assayed in mice as viability test. All the T. gondii isolates undergo molecular identification using PCR and genotyped utilizing nPCR/RFLP analysis of SAG2 gene. The total seropositivity of live sheep and goat was 47.15 and 39.2% utilizing ELISA and OTRT respectively. Concerning abattoirs, seropositivity, positive microscopic examination, mice viability from sheep samples were 47.1%, 37.3% and 44.1% respectively while that of goats were 45.5%, 33.3% and 48.6% respectively. Eighteen T. gondii isolates were affirmed utilizing PCR. Genotyping confirmed 10 isolates (55.5%) as type II, 6 (33.3%) as type III and 2 (11.1%) as atypical genotypes. Type II and III are the genotypes mostly circling among small ruminants in Egypt and this is most significance for the public health in Egypt.

New approach to differentiate primary from latent Toxoplasma gondii abortion through immunoglobulin and DNA interpretation.

BACKGROUND: Toxoplasma gondii is an acute or latent zoonotic abortifacient human protozoan. Women may be aborted due to recent or latent infection during pregnancy or order to flare up of the dormant bradyzoites to acute tachyzoites (latent opportunistic relapse). AIMS: 1) to validate the interpretation of IgM and IgG immunoglobulins seromonotoring with DNA comparative results in differentiating recent from latent T. gondii abortion. METHOD: Blood with the corresponding placental or uterine wash samples were collected from 73 aborted Egyptian women from Cairo and Giza labour wards. Patients aborted in any of the phases (Ph-1, Ph-2, Ph-3 and Ph-4 were corresponding to abortion at the 1st, 2nd and 3rd trimesters plus females who gave birth with congenital anomalies), respectively. All aborted patients were assayed serologically by Enzyme Linked Immunosorbent Assay (ELISA) for IgM and IgG titers and the compatible DNA from placenta and uterine wash tissues by conventional Polymerase Chain Reaction (PCR) specific for T. gondii. RESULTS: Sero-positive aborted women were 50.7% by ELISA versus 37% by PCR. Not all T. gondii sero-positive aborted women were having T. gondii DNA or harboring compatible placental T. gondii cysts. This denotes that immunoglobulins alone are insufficient criteria for confirming toxoplasma abortion. CONCLUSION: Immunoglobulins with DNA comparative results can possibly differentiate recent from latent T. gondii abortion at higher precision. We recommend the need for routine monitoring of T. gondii i.e. (pre-, during and post-delivery).

Prevalence and phylogenetic characterization of Listeria monocytogenes isolated from processed meat marketed in Egypt.

Because of its high case fatality rate, listeriosis locates among the most frequent causes of death due to food-borne illness. In this study, a total of 150 processed meat samples were collected from Giza Governorate, Egypt. Phenotypic and genotypic identification of Listeria monocytogenes was performed using PCR incorporating listeriolysin O virulence gene hlyA followed by DNA sequence analysis. L. monocytogenes was confirmed in 4% of each of beef burger, minced meat, and luncheon samples. Phylogenetic analysis showed that all the six Egyptian isolates have high homology with Colombian isolate (EF030606), except one Egyptian isolate which showed high homology with Indian isolate (EU840690). The public health significance of these pathogens as well as recommended sanitary measures were discussed.