Control of citrus molds using bioactive coatings incorporated with fungal chitosan/plant extracts composite.

BACKGROUND: The ongoing postharvest loss in citrus fruits, due to fungal infection, is a chronic economic and agricultural problem. Most of citrus damage is caused by Penicillium spp., e.g. green mold (P. digitatum) and blue mold (P. italicum). Fungal chitosan, from Mucor rouxii, and plant extracts from cress seeds, olive leaves, pomegranate peels and senna pods, were evaluated as antifungal agents against the phytopathogenic fungi, P. digitatum and P. italicum, using in vitro qualitative and quantitative assays. RESULTS: All natural agents tested exhibited potent antifungal activity; the most powerful agent was cress (Lepidium sativum) seed extract, followed by pomegranate (Punica granatum) peel extract. Fungal chitosan also had a remarkable fungicidal potentiality using both evaluation assays. Penicillium digitatum was generally more resistant than P. italicum toward all examined agents. The incorporation of each individual natural agent in coating material resulted in a great reduction in fungal growth and viability. The addition of chitosan combined with cress and pomegranate extracts, to the coating materials, prevented coated citrus fruit from decay by green and blue mold for a 2-week storage period. CONCLUSION: Natural derivatives could be recommended as powerful antifungal alternatives to protect citrus fruits from postharvest fungal decay.

Antifungal action of Pichia anomala against aflatoxigenic Aspergillus flavus and its application as a feed supplement.

BACKGROUND: The disastrous problem of animal feed contamination by mycotoxigenic fungi continues to challenge researchers and health overseers worldwide. With the aim of preventing Aspergillus flavus growth in vitro and in corn feed, the yeast Pichia anomala was examined as a biocontrol agent. RESULTS: The yeast strain could efficiently prohibit the growth of A. flavus. P. anomala was able to produce exo-chitinase and ß-1,3-glucanase which could be suggested as a mode of action for its antifungal activity. Scanning electron microscopy of cultured P. anomala with fungal hypha revealed that A. flavus was colonised by the biocontrol yeast which subsequently led to complete hyphal lysis and deterioration. The supplementation with of P. anomala cells, as a protein source, led to an obvious increase in animals' weight gain and protein content in feed grain. Moreover, after consumption of P. anomala-supplemented feed, there was a remarkable decrease in the mortality rate among fed animals. CONCLUSION: P. anomala could be strongly recommended as a biocontrol agent against A. flavus which contaminates animal feed. Furthermore, the application of yeast cells, as a feed additive, proved its efficiency for escalating protein content and enhancing animal performance.

Production of anticandidal cotton textiles treated with oak gall extract.

Candida albicans, one of the most dreadful fungal pathogens threatening humans, could not be easily prevented. The anticandidal activity of oak gall extract, Quercus infectoria (QIE), was investigated as a potential natural alternative to synthetic and chemical fungicides. QIE anticandidal potentiality was confirmed using both qualitative and quantitative assays. Cotton textiles were treated with QIE and then evaluated as anticandidal fabrics. QIE-treated textiles had a potent anticandidal activity, which could completely inhibit the inoculated C. albicans cells. The durability of anticandidal activity in QIE-treated textiles almost completely disappeared after the fourth laundering cycle. QIE could be recommended, however, as a potent anticandidal agent for preparing antiseptic solutions and emulsions and as a finishing agent for manufacturing anticandidal disposable diapers and hygienic clothes.

Surface decontamination and quality enhancement in meat steaks using plant extracts as natural biopreservatives.

Nine plant extracts were evaluated as biopreservatives to decontaminate and maintain the quality of meat steaks. Most of the extracts exhibited a remarkable antibacterial activity against antibiotic resistant strains from Salmonella Typhimurium and Staphylococcus aureus. The pomegranate peel extract (PPE), cinnamon bark extract (CBE), and lemon grass leaves extract (LGE) were the most effective as bactericides, with minimal inhibitory concentrations (MIC) of 250, 350, and 550 µg/mL, respectively. The most effective treatments, for decontaminating meat steak surfaces, were the application of combined PPE, CBE, and LGE at their MIC values and the treatment with double MIC from PPE; these treatments resulted in complete bacterial inhibitions during the first 2 days of storage period for 7 days. The sensory evaluation of treated steaks revealed that these two treatments had the highest panelist overall scores. The highest scores, for individual attributes, were observed in the treated steaks with double MIC from PPE. Application of plant extracts could be impressively recommended for comprehensive meat decontamination and quality attributes enhancement.

Brucella spp. infection in large ruminants in an endemic area of Egypt: cross-sectional study investigating seroprevalence, risk factors and livestock owner's knowledge, attitudes and practices (KAPs).

BACKGROUND: Brucellosis is regarded as one of the major zoonotic infections worldwide. It was first reported in Egypt in 1939 and is now endemic, the predominate species of Brucella in cattle and buffalo in Egypt is B. melitensis. The aim of the study was to estimate seroprevalence of Brucella spp. in cattle and buffalo reared in households in an Egyptian village, identify risk factors for animals testing seropositive and to assess the knowledge, attitudes and practices (KAPs) of livestock owners with regards to brucellosis. METHODS: A cross-sectional study was carried out in a village in Menufiya Governorate of Egypt. In June and July 2009, 107 households were selected using systematic sample and all lactating cattle and buffalo present in the household were sampled and tested for antibodies against Brucella spp. In addition, a questionnaire collecting information on potential risk factors for Brucella spp. infection in cattle and buffalo was administered to the household member responsible for rearing the livestock. Between December 2009 and February 2010 households were revisited and a second questionnaire regarding KAPs associated with brucellosis was administered. RESULTS: True individual and household seroprevalence were estimated to be 11.0% (95% CI: 3.06% to 18.4%) and 15.5% (95% CI: 6.61% to 24.7%), respectively. Cattle and buffalo kept in a household with sheep and goats had 6.32 (95% CI: 1.44 to 27.9) times the odds of testing seropositive for Brucella spp., compared to cattle and buffalo that were not. Most participants in the study stated that livestock owners assist in the parturition of ruminants without wearing gloves and that some farmers sell animals which they suspect are Brucella infected to butchers or at market. Many participants made their livestock's milk into cheese and other dairy products without pasteurising it. CONCLUSIONS: Brucellosis was endemic at high levels, in the current study. Although livestock owners had good general knowledge of brucellosis, they still appeared to participate in high-risk behaviours, which may contribute to the high seroprevalence in the area. Veterinarians, public health authorities and other community leaders need to collaborate to control the disease in animals and to manage the risk of human exposure.

Exploration of Islamic medicine plant extracts as powerful antifungals for the prevention of mycotoxigenic Aspergilli growth in organic silage.

BACKGROUND: Feed contamination with mycotoxins is a major risk factor for animals and humans as several toxins can exist as residues in meat and milk products, giving rise to carry-over to consumers via ingestion of foods of animal origin. The starting point for prevention, in this chain, is to eliminate the growth of mycotoxigenic fungi in the animal forage. Ten plant extracts, recommended in Islamic medicine, were evaluated as antifungal agents against mycotoxigenic Aspergilli, i.e. Aspergillus flavus and A. ochraceus, growth in organic maize silage. RESULTS: Most extracts had remarkable antifungal activities using both qualitative and quantitative evaluation methods. Cress (Lepidium sativum) seed extract was proven to be the most powerful among the plants examined. Blending of the most effective extracts (garden cress seed, pomegranate peel and olive leaf extracts), individually at their minimal fungicidal concentrations, with maize silage resulted in the reduction of inoculated A. flavus colony counts by 99.9, 99.6 and 98.7%, respectively, whereas silage blending with the combined extracts completely prohibited fungal growth for up to 30 days of incubation under aerobic conditions. CONCLUSION: Besides the health promoting effects, silage blending with the bioactive plant extracts examined could lead to the required protection from pathogenic and mycotoxigenic fungi.

Prevalence of intestinal parasitic infections and their associated risk factors among preschool and school children in Egypt.

Intestinal parasitic infections (IPIs) are among the major public health problems globally, particularly in developing countries like Egypt. This study aimed to evaluate prevalence and risk factors associated with IPIs among preschool and school children in Egypt. A cross-sectional study was conducted on 996 randomly selected preschool and school-aged children in Gharbia governorate during January to April 2018. Stool specimens were examined for the presence of the parasite by direct smear and the formol-ether concentration methods. The overall prevalence of IPIs was 46.2%. Entamoeba histolytica and Ascaris lumbricoides were the most predominant parasites (12.7% per each). This is followed by Enterobius vermicularis (8.6%), Giardia lamblia (7.1%), Cryptosporidium parvum (1.5%), Heterophyes heterophyes (1.4%), Hymenolepis nana (0.7%), Hookworms (0.6%), Fasciola hepatica (0.5%) and Dipylidium caninum (0.4%). Infected children with no symptoms (26.8%) were significantly (P < 0.001) more frequent than those with medical complaint (19.4%). Socio-demographic predictors of IPIs were preschool age (OR = 4.9; P < 0.001; 95%CI 3.3-7.3), living in rural dwellings (OR = 1.96; P < 0.001; 95%CI 1.5-2.5), and belonging to a low-income family (OR = 4.7; P < 0.001; 95%CI 2.3-9.3). The absence of safe drinking water, lack of hand washing (after soil contact, or before meals, or after toilet usage), and eating unwashed vegetables were risk factors for IPIs in the study region (OR = 1.3-6.9, P < 0.001 -P = 0.05). Higher odds for exposure to potential zoonotic parasites were evident in children with pets in their homes for G. lambia and D. caninum (OR = 2.1-8.3; P = 0.02 -P = 0.04), children having household reared ruminants for C. parvum (OR = 10.4; P < 0.001), and children that play with stray animals for E. histolytica and Hookworm (OR = 1.8-6.3; P = 0.04 -P = 0.05)compared to other children with no animal contact. The present study highlights the importance of periodic screening and treatment of IPIs in children, deworming companion animals, and public education for effective prevention of IPIs in children in Egypt.

Anticandidal activity of pomegranate peel extract aerosol as an applicable sanitizing method.

Pomegranate is a wonderful fruit from the paradise which contains a wide variety of precious phytochemical compounds applicable in the fields of therapeutics and health care. Candida albicans is the most common etiological agent for many clinical mycoses which could lead to human and animal death. Determination of the anticandidal activity of pomegranate peel extracts (PPE), and application of PPE aerosol as sanitizer agent against C. albicans contamination were investigated. Agar diffusion assay and broth microdilution susceptibility test were applied for qualitative and quantitative determining the PPE anticandidal activity, respectively, versus commonly used fungicides. Aerosolization of PPE using an experimentally designed sanitizer room was applied for examining C. albicans sanitation potentiality of extract. PPE exhibited potent anticandidal activity against C. albicans strains comparing with standard fungicides in both used susceptibility techniques. Methanol, ethanol and water extracts were the most effective for inhibiting C. albicans growth. PPE aerosol was an efficient method for complete sanitizing of semi-closed places against C. albicans growth. Application of PPE aerosol is a proper sanitizing method for preventing C. albicans contamination and growth in suspected places.

Selenium Nanoparticles and Spirulina Alleviate Growth Performance, Hemato-Biochemical, Immune-Related Genes, and Heat Shock Protein in Nile Tilapia (Oreochromis niloticus).

The present investigation aimed to evaluate the influence of selenium nanoparticles (Se-NPs) or/and spirulina (SP) on the growth, immunity, and oxidation resistance of Nile tilapia. Four groups of fish fed diets with Se-NPs or/and SP at 0 g (control), 1 g SP/kg diet (SP), 1 mg Se-NPs/kg diet (Se-NPs), and 1 g SP + 1 mg Se-NPs/kg diet (SP/Se-NPs) for 60 days. Fish fed Se-NPs or/and SP displayed significantly improved weight gain (WG) and decreased feed conversion ratio (P < 0.05). The highest WG has observed in fish fed both Se-NPs and SP, while the specific growth rate was improved by feeding Se-NPs only or both Se-NPs and SP (P < 0.05). Blood albumin was increased significantly with Se-NPs with regard to the control (P < 0.05), while there were no significant differences between fish fed Se-NPs or/and SP. Blood total protein also was improved by feeding Se-NPs only or both Se-NPs and SP (P < 0.05). Further, blood immunoglobulin M was increased by feeding both Se-NPs and SP (P < 0.05), while the differences were insignificantly differing with fish fed only Se-NPs (P > 0.05). The transcription of liver superoxide dismutase (SOD) and tumor necrosis factor-alpha (TNF-α) genes was upregulated significantly by Se-NPs or/and SP (P < 0.05). Interestingly, TNF-α was significantly upregulated by SP when compared to those fed Se-NPs only or both Se-NPs and SP. However, heat shock protein 70 (HSP70) gene transcription was downregulated by Se-NPs or/and SP (P < 0.05). Based on the measured parameters, the mixture of both Se-NPs and SP is highly recommended for the welfare of Nile tilapia.

Possibility of fighting food borne bacteria by egyptian folk medicinal herbs and spices extracts.

UNLABELLED: Phytotherapy (herbal medicine) have a long-standing history in Egypt. Current study investigated the antimicrobial potentialities of twenty five herbs and spices which are widely used in folk medicine by Egyptian housewives to treat gastrointestinal disorders against seven bacterial strains, mostly food borne including pathogens. They were tested by using paper disc diffusion technique as qualitative assay and agar dilution method for determining the minimum inhibitory concentration (MIC) of herbs extracts. Among screened plants, basil, cardamom, cinnamon, clove, lemon grass, mustard, oregano, rosemary, sage and thyme extracts exhibited notable antimicrobial activities against most of the tested strains. Cinnamon extract was the most inhibitor followed by clove, whereas extracts of chamomile, rose of Jericho, safflower and turmeric showed weak antibacterial activities against most of the tested strains. The most sensitive strain to plant extracts was B. subtilis and the most resistant strain was Ps. fluorescens. CONCLUSION AND RECOMMENDATIONS: herbs and spices extracts -used in Egyptian folk medicine for treating many gastrointestinal disorders - could be successfully applied as natural antimicrobials for elimination of food borne bacteria and pathogens growth.

A multifaceted risk model of brucellosis at the human-animal interface in Egypt.

Brucellosis is a highly contagious zoonosis affecting humans and a wide range of domesticated and wild animal species. An important element for effective disease containment is to improve knowledge, attitudes and practices (KAP) of afflicted communities. This study aimed to assess the KAP related to brucellosis at the human-animal interface in an endemic area of Egypt and to identify the risk factors for human infection. A matched case-control study was conducted at the central fever hospitals located in six governorates in northern Egypt. Face-to-face interviews with cases and controls were conducted using a structured questionnaire. In total, 40.7% of the participants owned farm animals in their households. The overall mean practice score regarding animal husbandry, processing and consumption of milk and dairy products were significantly lower among cases compared with controls (-12.7 ± 18.1 vs. 0.68 ± 14.2, respectively; p < .001). Perceived barriers for notification of animal infection/abortion were predominate among cases and positively correlated with participants' education. The predictors of having brucellosis infection were consumption of unpasteurized milk or raw dairy products and practicing animal husbandry. Applying protective measures against infection significantly reduced its risk. A model predicting risk factors for brucellosis among those who own animal showed that frequent abortions per animal increased the chance for brucellosis infection among human cases by 50-fold (95% CI: 8.8-276.9), whereas the use of protective measures in animal care reduced the odds (OR = 0.11 [95% CI: 0.03-0.45]). In conclusion, consumption of unprocessed dairy products was equally important as contact with infected/aborted animals as major risk factors for Brucella spp. infection among humans in Egypt. There is poor knowledge, negative attitudes and risky behaviours among villagers which can perpetuate the risk of brucellosis transmission at the human-animal interface. This supports the need for integrating health education into the national brucellosis control programme.

Update on the status of leptospirosis in New Zealand.

Leptospirosis is a zoonotic disease that poses public health and ecological threats worldwide. In New Zealand (NZ), incidence of the disease is relatively high compared to other developed countries. The aim of this review was to describe the epidemiological status, ecological risk of leptospirosis and prevention in NZ. Disease notification data for leptospirosis in humans in NZ from 2010 to 2015 were collected from the Institute of Environmental Science and Research database. These data were supported by a literature review of epidemiological studies in human and animal populations. During this six-year period, exotic serovars of Leptospira interrogans sv Australis and Leptospira kirschneri sv Grippotyphosa were identified in patients who had travelled abroad to Samoa (Pacific Ocean) and Thailand, respectively. Most cases of leptospirosis were recorded in New Zealanders of European ethnicity, followed by Maori people. Males had a nine-fold increased risk compared to females mostly due to occupation. The risk of leptospirosis increased gradually with the age, with a peak in the 40 to 49-year-old age group, after which it decreased. Workers in meat-processing and farming industries appeared at highest-risk of occupational exposure compared with other risky occupations such as hunters, veterinarians, technicians, stock truck drivers and lake workers. Other cases were also attributed to outdoor exposures or travelling overseas. Highest disease notification rates occurred in the West Coast region of the South Island (average annual incidence 9.7 per 100,000 people), followed by Whanganui region (8.2 per 100,000) and Hawke's Bay region (8 per 100,000) in the North Island. Vaccines currently available for animals are specific for cattle, sheep, deer and dogs and do protect against all serovars present in NZ. The development and use of molecular diagnostics is crucial for specific identification of Leptospira isolates and informing deployment of efficient vaccines.

The potentiality of cross-linked fungal chitosan to control water contamination through bioactive filtration.

Water contamination, with heavy metals and microbial pathogens, is among the most dangerous challenges that confront human health worldwide. Chitosan is a bioactive biopolymer that could be produced from fungal mycelia to be utilized in various applied fields. An attempt to apply fungal chitosan for heavy metals chelation and microbial pathogens inhibition, in contaminated water, was performed in current study. Chitosan was produced from the mycelia of Aspergillus niger, Cunninghamella elegans, Mucor rouxii and from shrimp shells, using unified production conditions. The FT-IR spectra of produced chitosans were closely comparable. M. rouxii chitosan had the highest deacetylation degree (91.3%) and the lowest molecular weight (33.2kDa). All chitosan types had potent antibacterial activities against Escherichia coli and Staphylococcus aureus; the most forceful type was C. elegans chitosan. Chitosan beads were cross-linked with glutaraldehyde (GLA) and ethylene-glycol-diglycidyl ether (EGDE); linked beads became insoluble in water, acidic and alkaline solutions and could effectively adsorb heavy metals ions, e.g. copper, lead and zinc, in aqueous solution. The bioactive filter, loaded with EGDE- A. niger chitosan beads, was able to reduce heavy metals' concentration with >68%, and microbial load with >81%, after 6h of continuous water flow in the experimentally designed filter.

Foodborne Pathogens Prevention and Sensory Attributes Enhancement in Processed Cheese via Flavoring with Plant Extracts.

Cheese contaminations with foodborne bacterial pathogens, and their health outbreaks, are serious worldwide problems that could happen from diverse sources during cheese production or storage. Plants, and their derivatives, were always regarded as the potential natural and safe antimicrobial alternatives for food preservation and improvement. The extracts from many plants, which are commonly used as spices and flavoring agents, were evaluated as antibacterial agents against serious foodborne pathogens, for example Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, and Escherichia coli O157:H7, using qualitative and quantitative assaying methods. Dairy-based media were also used for evaluating the practical application of plant extracts as antimicrobial agents. Most of the examined plant extracts exhibited remarkable antibacterial activity; the extracts of cinnamon, cloves, garden cress, and lemon grass were the most powerful, either in synthetic or in dairy-based media. Flavoring processed cheese with plant extracts resulted in the enhancement of cheese sensory attributes, for example odor, taste, color, and overall quality, especially in flavored samples with cinnamon, lemon grass, and oregano. It can be concluded that plant extracts are strongly recommended, as powerful and safe antibacterial and flavoring agents, for the preservation and sensory enhancement of processed cheese.

Risk of Toxocara canis eggs in stray and domestic dog hair in Egypt.

The aim of the study was to assess whether the hair of stray and domestic dogs in Egypt was contaminated with the eggs of the zoonotic parasite Toxocara canis, and also to identify risk factors for T. canis for contamination. Paired samples of hair and feces were collected from 53 stray and 47 domestic dogs, and hair samples were obtained from a further 11 stray and 9 domestic dogs. All samples were examined to identify T. canis eggs and, if eggs were found, their maturation stage. Eggs were identified in 26.6% of stray and 10.7% of domestic dog's hair samples. A significantly increased risk of embryonated T. canis eggs in hair samples was found in stray dogs (p=0.04), stray dogs had 3.18 (95% CI: 1.04-9.74) times the odds of having T. canis eggs present compared with domestic dogs. There was also a significant difference (p=0.02) between the mean quantity of eggs per gram in stray (77.6±6.54) and domestic (48.7±6.65) dog's hair. Fecal examination found a T. canis egg prevalence of 35.8% and 21.3% in stray and domestic dogs, respectively. As no domestic dogs which were positive from hair samples had negative fecal samples, this indicates that the presence of T. canis eggs in hair is probably due to self contamination. Two stray dogs had positive hair samples but negative fecal samples indicating that contamination may also be environmental. As both non-embryonated and embryonated T. canis eggs were found in the hair of domestic dogs, direct contact with dogs may be a potential risk factor for transmission of T. canis eggs to humans.

Infants exposure to aflatoxin M1 as a novel foodborne zoonosis.

Occurrence of aflatoxin M(1) (AFM(1)) in infant formula milk powder (IFMP) and maternal breast milk (MBM) was investigated as a risk factor affects the health of newborns in Egypt. A total of 125 IFMP and 125 MBM samples were collected and examined for the presence of AFM(1) using competitive ELISA test. The results indicated that the relative risk (RR) of exposure to AFM(1) via consumption of MBM was higher than IFMP (RR; 1.6, 95%CI; 1.28-2.03, p=0.0001). The mean concentrations of AFM(1) were significantly differed (p<0.0001) between MBM (74.413 ± 7.070 ng/l) and IFMP (9.796 ± 1.036 ng/l). High frequency distributions were detected within the range of 5-25 ng/l and >50-100 ng/l in IFMP and MBM, respectively. The average daily exposure of newborns to AFM(1) via consumption of MBM and IFMP was 52.684 and 8.170 ng, respectively, with a significant difference at p<0.0001. Consumption of raw milk by lactating mothers exhibited a significant correlation (p<0.0001) with the presence of AFM(1) in their milk. In conclusion, this work established a pioneering concept that AFM(1) may be considered as an etiological factor for a novel foodborne zoonosis identified as Aflatoxicosis M(1).

Antimicrobial textile treated with chitosan from Aspergillus niger mycelial waste.

The waste biomass of Aspergillus niger, following citric acid production, was used as a source for fungal chitosan extraction. The produced chitosan was characterized with deacetylation degree of 89.6%, a molecular weight of 25,000 dalton, 97% solubility in 1% acetic acid solution and comparable FT-IR spectra to standard shrimp chitosan. Fungal chitosan was applied as a cotton fabric finishing agent using pad-dry-cure method. The topographical structure of chitosan-treated fabrics (CTF) was much improved compared with control fabrics. CTF, after durability tests, exhibited a powerful antimicrobial activity against both E. coli and Candida albicans, the captured micrographs for E. coli cells contacted with CTF showed a complete lysis of cell walls with the prolonging contact time. The produced antimicrobial CTF could be proposed as a suitable material for many medical and hygienic applications.

Cross-species surveillance of Leptospira in domestic and peri-domestic animals in Mahalla City, Gharbeya Governorate, Egypt.

A survey of 179 animals (black rats, dogs, sheep, buffaloes, cattle, donkeys, weasels, and cats) for Leptospira infection was conducted in Mahalla City (Lower Egypt). Blood, urine, and kidney were collected and tested by culture, microscopic agglutination test (MAT), and/or polymerase chain reaction (PCR). Among rats, 26% were positive by PCR, including 7% that were also positive by culture for L. interrogans serovars Grippotyphosa, Pyrogenes, and Icterohaemorrhagiae. L. borpetersenii serovar Polonica was isolated for the first time in Egypt in three rats. MAT titers ≥ 1:800 were observed in 11% of rats and 12% of dogs. L. interrogans serovar Grippotyphosa was detected in one cat. Sheep and donkeys were negative for leptospirosis by all methods. Buffaloes and cattle were seropositive in 20% and 44% of animals, respectively. Data indicate that several pathogenic serovars are circulating in the animals, which may pose exposure risks and account for high rates of acute febrile illness.

Potential zoonotic pathways of Salmonella enteritidis in laying farms.

Salmonella Enteritidis is a communicable zoonotic bacterium. The present investigation was done to evaluate the potential occurrence of Salmonella Enteritidis in laying hen farms and its contamination pathways. Samples were collected from 10 laying hen farms located in the Delta of Egypt. Cloacal swabs (n=300), eggshell swabs (n=400), and hand swabs from egg packagers (n=38) were collected. Pools of ovary and oviduct were obtained from 150 hens; all samples were examined for the presence of Salmonella Enteritidis. Results indicated that Salmonella Enteritidis may initially occupy in ovary, oviduct, and cloaca. The risk for eggshell contamination is highly impending from laying hen flocks infected with Salmonella Enteritidis with percentage of >30%. Farms having eggshell contamination percentage of >60% with Salmonella Enteritidis provided a high risk for packagers' hand contamination. Questionnaire of egg packagers specified that seven out of the eight smoker packagers suffered from repeated Salmonellosis. Thus, smoking during egg packing process could be considered as an exposure factor to contract the infection via hand-cigarette-mouth route.

Brucella infection in fresh water fish: Evidence for natural infection of Nile catfish, Clarias gariepinus, with Brucella melitensis.

Brucellosis is endemic among ruminants in the Nile Delta region of Egypt, where recent reports suggest that the incidence of human infection is increasing. In this region the practice of throwing animal waste into Nile canals is common. As a result, water can be contaminated with potential zoonotic pathogens such as B. melitensis that could infect fish. This study aimed at isolating and characterizing B. melitensis from Nile catfish. Serum samples from 120 catfish captured from Nile canals and 120 farmed catfish were tested for the presence of antibodies against Brucella spp. by using the Rose Bengal Test (RBT) and the Rivanol test (Riv T). Skin swabs from all fish and samples from internal organs (liver, kidney and spleen) from all serologically positive fish were cultured to identify B. melitensis biovar 3 isolates. Polymerase Chain Reaction (PCR) was used to confirm the results. 9.2% and 8.3% of serum samples from Nile catfish were positive by RBT and Riv T, respectively. None of the samples from farmed catfish were seropositive. B. melitensis biovar 3 was isolated from 5.8%, 4.2%, 5.8% and 13.3% of liver, kidney and spleen samples and skin swabs, respectively. To our knowledge this is the first report of isolation of B. melitensis biovar 3 from fresh water fish. Our results suggest that Nile catfish are naturally infected with B. melitensis biovar 3 and this may play a role in the epidemiology of brucellosis. The public should be aware of the consequences of disposing of animal waste into the canals and public health authorities should consider the potential role of catfish as a source of infection.