Despite their importance, only few studies focused on the mosquitoes borne microbial diseases, especially bacterial and fungal diseases, their vectorial capacity toward microorganisms, and their important role in raw milk contamination with pathogens in some unsanitary dairy farms. In Egypt, where Culex pipiens is the historical main vector of lymphatic filariasis, only few studies discussed the isolation of pathogens from the midguts of different stages of C. pipiens. This study aims to isolate and identify the pathogenic symbiotic microorganisms inside the midgut of adult female C. pipiens as well as investigate its ability to transmit their midgut pathogens to raw milk. A total of 750 field strain C. pipiens larvae of the second and third larval instars were collected from ponds water around the livestock farms in Mariotteya, Giza, Egypt, for microbial pathogen isolation and identification. All collected larval instars were transported to the laboratory at the Research and Training Center on Vectors of Diseases (RTC), Ain Shams University, where they were maintained for further studies. Six groups of C. pipiens were tested for the incidence of various pathogenic microorganisms in their midguts and their possibility to contaminate commercial sterilized milk. Traditional PCR assays and sequencing method detected and identified 16srRNA genes of the predominant hemolytic isolates from milk and midguts of female C. pipiens. The phylogenetic analyses of the obtained isolates were performed based on NCBI data. Three strains of Bacillus anthracis strain CPMESA 2021, Staphylococcus warneri strain CPSAME 2021, and Bacillus cereus strain CPSEMA 2021, which represent most food pathogens, were found in the midguts of C. pipiens and were submitted to the GenBank database with the accession numbers OK585071, OK576651, and OK585052, respectively. The isolation of these strains from mosquitoes raises contemporary issues concerning milk safety, such as bacterial isolates, the degree of the vectorial capacity of mosquitoes, milk production and processing conditions, and human pathogenicity. Such serious issues need further investigation.

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The aim of this research was to study the utilization ability of the salt whey and milk permeate to produce exopolysaccharides (EPSs) from halophilic bacteria. Where cheese whey is simultaneously an effluent with nutritional value and a strong organic and saline content. However, it is drained in the sewers, the EPSs are highly heterogeneous polymers produced by different species of bacteria and have recently been attracting considerable attention from biotechnologists because of their potential applications in many fields. Thus, we have isolated some halophilic bacteria that showed the ability to produce EPS from whey and milk permeate. A total of 46 strains of moderately halophilic bacteria were isolated from two types of samples. The first type was dairy samples (Baramily cheese whey and mish cheese), while the second type was water samples (salty lake water and its sediments) of Wadi El-Natrun Valley (lakes Hamra, UmRisha, and Baida), Beheira Governorate, Egypt. From isolated strains there are twelve strains were having the ability to produce exopolysaccharides but only seven strains can produce EPS from whey and milk permeate. The growth conditions i.e. concentrations of NaCl, pH value and different incubation temperature, of isolates were determined. The effect of these conditions on the production of EPS was investigated. The obtained results indicated that the optimum conditions for the production of EPS by these strains were 10 % NaCl, pH 7 and the optimum incubation temperature was 37°C. Three strains showed the highest production of exopolysaccharides. These strains were identified using two methods the first method was biolog system and the second one was 16S rRNA sequence analysis method. It could be identified as Alteribacillus bidgolensis and Bacillus licheniformis. Alteribacillus bidgolensis (strain P4B) produced the highest amount of EPS (52 g/L) from whey followed by Bacillus licheniformis (DSM 13) (42 g/L), while the highest amount of EPS produced from milk permeate was (43 g/L) by Alteribacillus bidgolensis (strain P4B) followed by Bacillus licheniformis (DSM 13) (36 g/L).