Incidence of Cronobacter sakazakii in Dairy-based Desserts

Cronobacter sakazakii is one of emerging foodborne pathogens around the world. A total of 90 dairy-based desserts samples (ice cream, Muhallabia and rice pudding) were examined for detecting C. sakazakii. All samples were submitted for bacteriological examination and confirmed by molecular identification using 16S rRNA gene for C. sakazakii. The bacteriological and molecular examination revealed that the incidence of occurrence of C. sakazakii was 5.55% from the total dairy-based desserts samples, the highest percentage occurred in rice pudding samples (10%), while the incidence of C. sakazakii in ice cream and Muhallabia were 3.33% for each type. The results pointed out that high risk for human may occur by contaminated dairy-based desserts. The hygienic precautions must be taken during the processing of these types of products

Occurrence of Toxoplasma gondii in raw goat, sheep, and camel milk in Upper Egypt

Background and Aim: Toxoplasmosis is a worldwide zoonotic disease with harmful effects on animal and human health. Ingestion of contaminated raw milk has been suggested as a vehicle for transmission of Toxoplasma gondii to human. 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 (enzyme-linked immunosorbent assay [ELISA] and quantitative polymerase chain reaction [qPCR]). Materials and Methods: This study was conducted to determine the T. gondii prevalence using ELISA and qPCR in raw goat, sheep, and camels milk (30 samples for each) collected from different locations in Upper Egypt. Results: T. gondii IgG antibodies were detected in 90.0, 60.0, and 3.33% of goat, sheep, and camel milk samples, respectively. From the positive samples of T. gondii IgG, the parasitic DNA was detected only in two examined milk samples, one of them was present in goat milk sample and another one was found in sheep milk sample. On the other hand, the parasite was not detected in camels' milk samples. Conclusion: These results concluded that the raw milk was contaminated by T. gondii tachyzoites which could be a source of human infection. Restricted hygienic programs should be implemented in the animal farms to decrease the risk of milk contamination by this parasite

Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

Background and Aim: Milk adulteration is pivotal because it leads to worse effects in public health as human adverse reactions with clinical signs ranged from gastrointestinal signs to anaphylactic shock. This study was carried out to estimate the prevalence of adulteration in buffalo's milk sold in Assiut City, Egypt. Materials and Methods: A total of 50 raw buffalo's milk samples were collected and examined for adulteration by addition of cow's milk. The examination carried out by applying polymerase chain reaction-restriction fragment length polymorphism technique using cytochrome b (cyt b) gene primers and Hinf I enzymes. The size of target gene was 360 bp in both animal species and amplicon can be digested using Hinf I enzyme, this restriction enzyme divided the essential band to clear three bands at 360, 210, and 150 bp in cows' milk, while, the enzyme could not be cleaved the amplicon in buffalo's samples. Results: The obtained results cleared that the incidence of adulteration of buffalo's milk very high percentage reaches 90%. Conclusion: It could be concluded that the raw buffalo's milk sold in Assiut City subject to fraudulent practice and thus can lead to public health hazards

Prevalence of enterotoxins and other virulence genes of Staphylococcus aureus caused subclinical mastitis in dairy cows

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

Molecular diagnosis and biochemical studies of tick-borne diseases (anaplasmosis and babesiosis) in Aberdeen Angus Cattle in New Valley, 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

Prevalence and virulence factor genes of Bacillus cereus isolated from milk and some dairy products

Two hundred and fifty samples (Marketable milk, Ras cheese, Domiati cheese and ice cream), were randomly obtained from various dairy shops, supermarkets and ice cream shops in Assiut City, Egypt, during the period of November 2022 to March 2023. These samples were examined for the detection and counting of Bacillus cereus. The positive B. cereus isolates were analyzed by PCR to detect 16s rRNA gene for B. cereus, cytotoxin K (cyt K) and emetic gene (ces). Out of the 250 examined samples, 14 (5.6%) were contaminated by B. cereus. The analyzed ice cream had the highest prevalence (11.53%), followed by Ras cheese samples. The prevalence of ces gene recorded in the examined isolates was 57.14%, while cyt K gene was found in all isolates. Moreover, proteolytic and lipolytic activity investigations revealed that nearly all strains released protease enzyme (92.85%), most of which (64.28%) could produce lipase enzyme

Prevalence of Campylobacter Spp. in Marketable Milk and Some Milk Products in New Valley Governorate, Egypt

The current study aimed to determine the prevalence of Campylobacter in milk and milk products as well as the determination of isolated Campylobacter in Egypt's New Valley Governorate. 150 random samples of marketable milk (dairy farm and dairy shop) and some milk products: soft cheeses (Kareish, Domiati) and Ice cream (30 for each) were obtained from various locations in the New Valley Governorate. Campylobacter spp. were found in 6.6% of marketable milk from dairy farms. In addition, Campylobacter spp. were found in 3.3% of soft cheese samples (Kareish cheese). while they couldn't be detected in Domiati cheese samples. Moreover, Campylobacter spp. were found in 10% of ice cream samples. Campylobacter organisms were identified as C. coli (2%) and C. jejuni (2%) in marketable milk (dairy farm and dairy shop) and some milk products. In conclusion, Campylobacter species is detected in some milk and some milk products in New Valley governorate. So, restricted milk hygiene must be applied during milking, manufacturing, and marketing milk and its products.   

Detection of aflatoxins and novel simple regimes for their detoxification in milk and soft cheese

The purpose of this study was to detect AFs in milk and in the most popular cheese samples produced that were sold in New Valley governorate, Egypt. Trials were also carried out to find a simple, available way to counteract AFs in these products. Sixty samples of marketable milk and soft cheese (locally manufactured) were randomly collected from El-kharja markets, New Valley governorate, Egypt. Aflatoxin (AFs) was detected quantitatively in the samples by using enzyme linked immunosorbent assay (ELISA) analysis. The positive results (tainted with AFs) indicated that 100% of the examined samples have level of Afs that exceed the allowable limit. Furthermore, locally manufactured soft cheese had higher AFs contamination than marketable milk. Additionally, the experimental trials to treat contaminated milk with microwave heating, Mish contaminated samples with lemon, and Kareish cheese with carbonated water revealed 9.4, 43.9, and 54.9% decline in Afs levels, respectively. It could be concluded that examined milk and milk products are frequently contaminated with AFs which exceed the allowable limits and every regimen that was employed to treat the contaminated specimen was helpful

Prevalence of Some Spore Forming Food Poisoning Bacteria in Milk and Some Milk Products

Spore-forming bacteria are a group of bacteria can form spore and they grow aerobically and anaerobically as Bacilli and Clostridia spp. This group of bacteria has public health hazards and economic loss significance. This study was designed to determine the prevalence of spore-forming bacteria isolated from marketable milk and some dairy products as pasteurized milk, UHT milk, milk powder, and baby food (30 samples of each). The samples were purchased randomly from different dairy shops, supermarkets, and pharmacies in Assiut Governorate, Egypt. Bacillus cereus was detected in 23.33, 13.33, 6.66, 13.33, and 10% from marketable milk, pasteurized milk, UHT milk, milk powder, and baby food, respectively. While, Clostridium perfringens was recovered in 20, 6.66, 0, 3.33 and 0%, respectively. This study concluded that there is a need for hygienic measures must be applied in the milk and dairy products production and manufacture to minimize the possibility of entering the spore-forming bacteria in these products

Detection of Virulence and β-lactamase resistance genes of non-typhoidal Salmonella isolates from human and animal origin in Egypt "one health concern"

Abstract Background Non-typhoidal Salmonella (NTS) is a major foodborne zoonotic pathogen worldwide. In the current study, Various NTS strains were isolated from (cows, milk and dairy products in addition to humans) in New Valley and Assiut Governorate, Egypt. NTS were firstly serotyped and tested by antibiotic sensitivity test. Secondly, some virulence genes and Antibiotic resistance genes have been identified by using PCR. Finally, Phylogenesis was performed depending on the invA gene, for two S. typhimurium isolates (one of animal origin and the other of human origin for evaluating zoonotic potential). Results Out of 800 examined samples, the total number of isolates was 87 (10.88%), which were classified into 13 serotypes, with the most prevalent being S. Typhimurium and S. enteritidis. Both bovine and human isolates showed the highest resistance to clindamycin and streptomycin, with 90.80% of the tested isolates exhibiting MDR. The occurrence of the invA gene was 100%, while 72.22%, 30.56%, and 94.44% of the examined strains were positive for stn, spvC, and hilA genes, respectively. Additionally, blaOXA-2 was detected in 16.67% (6/ 36) of the tested isolates, while blaCMY-1 was detected in 30.56% (11of 36) of the tested isolates. Phylogenesis revealed a high degree of similarity between the two isolates. Conclusions The high occurrence of MDR strains of NTS in both human and animal samples with high degree of genetic similarity, shows that cows, milk and milk product may be a valuable source of human infection with NTS and interfere with treatment procedures