The present study was performed for the detection of T. gondii in raw milk samples of goat, sheep, and camel in Upper Egypt using two different techniques ELISA and qPCR

Background and Aim: Milk production is one of the main props for the national economy. One of the crucial problems in this industry is subclinical mastitis, which harms this industry that considered the backbone of the economy. It is an infectious and zoonotic disease; the infection can spread between dairy animals through milkers' hands, and milking machines, while the human infection occurs due to the consumption of apparently hygienic milk. Staphylococcus aureus is one of the main causative agents of clinical and subclinical mastitis. It is also considered one of the bacteria incriminated in food intoxication of humans due to its virulence factors as enterotoxins and toxic shock syndrome. The current study was designed to assess the prevalence of S. aureus and its enterotoxins, as well as, its other virulence factors in milk collected from cows that suffer from subclinical mastitis. Materials and Methods: Sixty cows were collected from different dairy farms located in Assiut Governorate, Egypt. These cows were subjected to the clinical examination of the udder and its lymph nodes before sampling. Milk samples were collected from clinically healthy udders. All the milk samples were examined by California mastitis test (CMT), polymerase chain reaction (PCR), and enzyme-linked immunosorbent assay (ELISA) for confirmation subclinical mastitis, presence of S. aureus and its enterotoxins genes and other virulence factors in the examined milk samples. Results: The cows included in the current study had healthy udders. The sixty collected milk samples were tested by CMT. 48/60 (80.0%) were positive samples; from the 48 positive samples, 46 (95.83%) samples were confirmed positive by S. aureus 16s rRNA PCR assay. Multiplex PCRs confirmed the presence of staphylococcus enterotoxin gene C (sec) in one sample, staphylococcus enterotoxin gene D (sed) in 23 samples, while ELISA assay confirmed the presence of the same enterotoxin in only two samples. On the other hand, other groups of genes responsible for some other virulence factors of S. aureus like the extracellular thermostable nuclease (nuc) gene were found in 33 samples, while toxic shock syndrome (tsst) gene and methicillin restraint S. aureus (mecA) gene were not detected in this study. Conclusion: Subclinical mastitis is one of the hidden factors that adversely affect the health of both animals and humans. The milk is usually appeared good and may be consumed by humans especially children; however, it causes severe public health problems. In addition, the infected animals with this form of mastitis can spread the infection to other dairy animals and may be turned to a clinical case of contagious mastitis that may be ended by animal culling or death. S. aureus is one of the main causes of subclinical mastitis in cattle. In addition to extracellular thermostable nuclease (nuc) gene, staphylococcus enterotoxin gene C (sec) and staphylococcus enterotoxin gene D (sed) are the most common virulence genes confirmed in subclinical mastitis milk. These results highlighted the need to apply more hygienic measures in the dairy farms to avoid spreading the infection between animals to ensure the production of safe and healthy food to humans.

Listeria monocytogenes is one of the most important emerging foodborne pathogens. The objectives of this work were to investigate the incidence of Listeria spp. and L. monocytogenes in soft cheese and ice cream in Assiut city, Egypt, and to examine the effect of some probiotic Bifidobacterium spp. (Bifidobacterium breve, Bifidobacterium animalis, or a mixture of the two) on the viability of L. monocytogenes in soft cheese. The existence of Listeria spp. and L. monocytogenes was examined in 30 samples of soft cheese and 30 samples of ice cream. Bacteriological analyses and molecular identification (using 16S rRNA gene and hlyA gene for Listeria spp. and L. monocytogenes, respectively) were performed on those samples. Additionally, Bifidobacterium spp. were incorporated in the making of soft cheese to study their inhibitory impacts on L. monocytogenes. Out of 60 samples of soft cheese and ice cream, 25 samples showed Listeria spp., while L. monocytogenes was found in only 2 soft cheese samples. Approximately 37% of soft cheese samples (11 out of 30) had Listeria spp. with about 18.0% (2 out of 11) exhibiting L. monocytogenes. In ice cream samples, Listeria spp. was presented by 47% (14 out of 30), while L. monocytogenes was not exhibited. Moreover, the addition of B. animalis to soft cheese in a concentration of 5% or combined with B. breve with a concentration of 2.5% for each resulted in decreasing L. monocytogenes efficiently during the ripening of soft cheese for 28 d. Listeria spp. is widely found in milk products. Probiotic bacteria, such as Bifidobacterium spp., can be utilized as a natural antimicrobial to preserve food and dairy products.

This study was carried out to estimate the prevalence of adulteration in buffalo?s milk sold in Assiut City, Egypt

Background and Aim: Anaplasmosis and babesiosis are tick-borne diseases that threaten livestock production with subsequent considerable economic losses. This study was conducted to diagnose Anaplasma and Babesia infection using molecular techniques in imported Aberdeen Angus cattle imported from Uruguay to El-Kharga Oasis in New Valley, Egypt, and to investigate the effects of disease on some serum biochemical and oxidative stress parameters. Materials and Methods: Blood samples were collected from 31 cattle, 21 diseased and ten apparently normal, of varying ages and sex. The blood was used for the preparation of blood smears, polymerase chain reaction assay, and separation of serum for biochemical investigation. The experimental production farm at the Faculty of Agriculture, New Valley University, was infested with ticks and variable clinical manifestations during the period from December 2017 to March 2018. One calf died of a suspected blood parasite infection. Results: The blood film examination revealed infection by blood parasites in 21 samples. Anaplasma marginale and Babesia bovis were identified in 12 and 14 samples, respectively. A total of 14 samples were examined by polymerase chain reaction (PCR) to make these identifications. Biochemical parameters showed significantly elevated serum alanine aminotransferase, aspartate aminotransferase, total bilirubin (T. Bil), and urea in blood from parasite-infected female cattle and male calves compared with controls. Increased serum total protein, globulin, and creatinine were recorded only in infected female cattle. The blood glucose level was significantly decreased in infected female cattle and male calves compared with controls. Furthermore, albumin and albumin/globulin ratio was significantly reduced in the infected female cattle. Oxidative stress profiles of infected animals showed a significant increase in serum nitric oxide and malondialdehyde, and both total antioxidant capacity and reduced glutathione (GSH) were significantly reduced in comparison with control animals. Conclusion: The incidence of A. marginale and B. bovis infection is high in imported Aberdeen Angus cattle in New Valley Province. PCR methods provide a short-term assessment of disease. An extensive epidemiological survey, employing serology together with molecular genetic methods, monitoring of abundance and distribution of tick vectors, availability of vaccination programs, and tracking of animal transport is also needed for control of blood parasites.

Background: L-asparaginase has been a widely employed as antitumor enzyme for the treatment of acute lymphoblastic leukemia for almost three decades. The enzyme takes advantage of the inability of tumor cells to synthesize the L-asparagine and is killed by L‐asparagine deprivation. Despite the availability of bacterial sources for L-asparaginase, there is a growing interest in identifying new microbial sources with improved therapeutic properties. Therefore, this study aims to investigate the production of L-asparaginase from a fungal source, to explore its potential as a novel alternative enzyme for cancer treatment. Results: Fusarium falciforme AUMC 16563 was used to produce L-asparaginase (123.42 U/mL) after 5 days, 0.2% glucose and 1.0% asparagine; were used at 25 ˚C and pH 8.0. Employing two columns of chromatography (DEAE-cellulose and Sephacryl S 200 HR), the enzyme was purified 14.26-fold, reaching a maximum activity of 5109.4 U/mg. SDS-PAGE revealed a 46.06 kDa asparaginase. The K _m and V _max values for pure asparaginase using asparagine was 5.77 × 10^− 2 mM and 128.22 µmol/min. Additionally, Fusarium falciforme AUMC 16563’ pure asparaginase demonstrated anticancer activity against PC-3 (a prostate cell line) with an IC₅₀ of 78.6 µg/mL, HePG-2 (a human hepatocellular carcinoma cell line) with an IC₅₀ of 69.6 µg/mL, HCT-116 (a colon cell line) with an IC₅₀ of 51.5 µg/mL and MCF-7 (a breast cancer cell line) with an IC₅₀ of 32.8 µg/mL. The expression levels of proapoptotic genes (BAX and p53) were significantly greater in the breast cancer cell lines treated with asparaginase than in the negative control breast cancer cell lines.The degree of DNA fragmentation in MCF-7 cells treated with Fusarium falciforme 16563’ pure asparaginase was 27.2 ± 0.69%, and that in MCF-7 cells treated with the drug Doxorubicin 24.1 ± 0.86% was significantly greater than that in the corresponding negative control cells 9.1 ± 1.01%. Finally, the biochemical profiles revealed no impact on the liver or the kidneys. These results suggested that asparaginase had relatively little effect on liver function. All hematological parameters were within normal range during the experiment. Conclusions: The results of the present study revealed a potent L-ASNase from endophytic F. falciforme isolated from Trifolium alexandrinum, which performs well under a variety of environmental circumstances and can be used in a number of commercial applications.