Genetic diversity and phylogenetic relationships of Clostridium perfringens strains isolated from mastitis and enteritis in Egyptian dairy farms.

BACKGROUND: Clostridium perfringens, a common environmental bacterium, is responsible for a variety of serious illnesses including food poisoning, digestive disorders, and soft tissue infections. Mastitis in lactating cattle and sudden death losses in baby calves are major problems for producers raising calves on dairy farms. The pathogenicity of this bacterium is largely mediated by its production of various toxins. RESULTS: The study revealed that Among the examined lactating animals with a history of mastitis, diarrheal baby calves, and acute sudden death cases in calves, C. perfringens was isolated in 23.5% (93/395) of the total tested samples. Eighteen isolates were obtained from mastitic milk, 59 from rectal swabs, and 16 from the intestinal contents of dead calves. Most of the recovered C. perfringens isolates (95.6%) were identified as type A by molecular toxinotyping, except for four isolates from sudden death cases (type C). Notably, C. perfringens was recovered in 100% of sudden death cases compared with 32.9% of rectal swabs and 9% of milk samples. This study analyzed the phylogeny of C. perfringens using the plc region and identified the plc region in five Egyptian bovine isolates (milk and fecal origins). Importantly, this finding expands the known data on C. perfringens phospholipase C beyond reference strains in GenBank from various animal and environmental sources. CONCLUSION: Phylogenetic analyses of nucleotide sequence data differentiated between strains of different origins. The plc sequences of Egyptian C. perfringens strains acquired in the present study differed from those reported globally and constituted a distinct genetic ancestor.

Escherichia coli and their potential transmission of carbapenem and colistin-resistant genes in camels.

BACKGROUND: Camels harbouring multidrug-resistant Gram-negative bacteria are capable of transmitting various microorganisms to humans. This study aimed to determine the distribution of anti-microbial resistance among Escherichia coli (E. coli) isolated from the feces of apparently healthy camels in Egyptian abattoirs. Additionally, we sought to characterize Shiga toxin-producing E. coli (STEC) strains, assess their virulence potential, and investigate the possibility of camels spreading carbapenem- and colistin-resistant E. coli. METHODS: 121 fecal swaps were collected from camels in different abattoirs in Egypt. Isolation and identification of E. coli were performed using conventional culture techniques and biochemical identification. All isolates obtained from the examined samples underwent genotyping through polymerase chain reaction (PCR) of the Shiga toxin-encoding genes (Stx1 and Stx2), the carbapenemase-encoding genes (blaKPC, blaOXA-48, blaNDM, and blaVIM), and the mcr genes for mcr-1 to mcr-5. RESULT: Bacteriological examination revealed 75 E. coli isolates. PCR results revealed that one strain (1.3%) tested positive for Stx1, and five (6.6%) were positive for Stx2. Among the total 75 strains of E. coli, the overall prevalence of carbapenemase-producing E. coli was 27, with 7 carrying blaOXA48, 14 carrying blaNDM, and 6 carrying blaVIM. Notably, no strains were positive for blaKPC but a high prevalence rate of mcr genes were detected. mcr-1, mcr-2, mcr-3, and mcr-4 genes were detected among 3, 2, 21, and 3 strains, respectively. CONCLUSION: The results indicate that camels in Egypt may be a primary source of anti-microbial resistance (AMR) E. coli, which could potentially be transmitted directly to humans or through the food chain.

Injectable systems of chitosan in situ forming composite gel incorporating linezolid-loaded biodegradable nanoparticles for long-term treatment of bone infections.

The objective of the current study was to create an efficient, minimally invasive combined system comprising in situ forming hydrogel loaded with both spray-dried polymeric nanoparticles encapsulating linezolid and nanohydroxyapatite for local injection to bones or their close vicinity. The developed system was designed for a dual function namely releasing the drug in a sustained manner for long-term treatment of bone infections and supporting bone proliferation and new tissues generation. To achieve these objectives, two release sustainment systems for linezolid were optimized namely a composite in situ forming chitosan hydrogel and spray-dried PLGA/PLA solid nanoparticles. The composite, in situ forming hydrogel of chitosan was prepared using two different gelling agents namely glycerophosphate (GP) and sodium bicarbonate (NaHCO3) at 3 different concentrations each. The spray-dried linezolid-loaded PLGA/PLA nanoparticles were developed using a water-soluble carrier (PVP K30) and a lipid soluble one (cetyl alcohol) along with 3 types of DL-lactide and/or DL-lactide-co-glycolide copolymer using nano-spray-drying technique. Finally, the optimized spray-dried linezolid nanoparticles were incorporated into the optimized composite hydrogel containing nanohydroxy apatite (nHA). The combined hydrogel/nanoparticle systems displayed reasonable injectability with excellent gelation time at 37 °C. The optimum formulae sustained the release of linezolid for 7-10 days, which reveals its ability to reduce the frequency of injection during the course of treatment of bones infections and increase the patients' compliance. They succeeded to alleviate the bone infections and the associated clinical, biochemical, radiological, and histopathological changes within 2-4 weeks of injection. As to the state of art in this study and to the best of our knowledge, no such complete and systematic study on this type of combined in situ forming hydrogel loaded with spray-dried nanoparticles of linezolid is available yet in literatures.

Pathways of Methicillin-Resistant Staphylococcus aureus in Animal Model: New Insights Regarding Public Health.

BACKGROUND: Staphylococcus aureus is considered one of the major threats regarding food safety worldwide. Methicillin-resistant S. aureus (MRSA) strains in livestock, companion animals, and wild animals continue to be a potential risk to people working with them. AIM: The current research aims to investigate the potential pathways of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) strains in the body after oral infection using the experimental mouse model. METHODS: Seven groups of SPF male mice were purchased and housed. On day 1, six groups of mice were infected orally by the sterile gastric probe using 100 µL/mice of LA-MRSA bacterial suspension (1 × 108 colony-forming units (CFU)/mL). The remaining group was kept as negative controls. Over 15 days, these animals have been monitored. Fresh fecal samples were screened for LA-MRSA at day 0, day 7 and day 14 following oral administration of MRSA strains. All animals were sacrificed at day 15, and internal organs (liver, lung, kidney, and intestine) were harvested aseptically and divided into two sections. The first part was histopathologically investigated, while the other half has been tested for LA-MRSA re-isolation. RESULT: The oral challenge of mice by MRSA strains showed that MRSA was re-isolated from feces and intestines of all inoculated mice groups and from internal organs (liver, lung, kidney and intestine) of most mice. Results were confirmed by the detection of the bacteria in gram-stained tissue sections and changes in H&E-stained histopathological tissue sections from these organs. CONCLUSION: Data from the present study indicate the possible colonization of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in internal organs following oral infection and thus posing a risk for food-borne infection of MRSA. Infected animals could pass LA-MRSA through feces again, resulting in increased dispersion and environmental contamination.

Emergence of ß-lactamase- and carbapenemase- producing Enterobacteriaceae at integrated fish farms.

BACKGROUND: Epidemiological studies suggested that determinants for antibiotic resistance have originated in aquaculture. Recently, the integrated agriculture-aquaculture system has been implemented, where fish are raised in ponds that receive agriculture drainage water. The present study aims to investigate the occurrence of ß-lactamase and carbapenemase-producing Enterobacteriaceae in the integrated agriculture-aquaculture and the consequent public health implication. METHODS: Samples were collected from fish, fishpond water inlets, tap water, outlet water, and workers at sites of integrated agriculture-aquacultures. Samples were also taken from inhabitants of the aquaculture surrounding areas. All samples were cultured on MacConkey agar, the Enterobacteriaceae isolates were tested for susceptibility to cephalosporins and carbapenems, and screened for blaCTX-M-15, blaSHV, blaOXA-1, blaTEM, blaPER-1, blaKPC, blaOXA-48, and blaNDM. Strains having similar resistance phenotype and genotype were examined for the presence of Incompatible (Inc) plasmids. RESULTS: A major proportion of the Enterobacteriaceae isolates were resistant to cephalosporins and carbapenems. Among the 66 isolates from fish, 34 were resistant to both cephalosporin and carbapenem groups, 26 to carbapenems alone, and 4 to cephalosporins alone. Of the 15 isolates from fishpond water inlets, 8 showed resistance to both groups, 1 to carbapenems alone, and 5 to cephalosporins alone. Out of the 33 isolates from tap water, 17 were resistant to both groups, and 16 to cephalosporins alone. Similarly, of the 16 outlet water isolates, 10 were resistant to both groups, and 6 to cephalosporins alone. Furthermore, of the 30 examined workers, 15 carried Enterobacteriaceae resistant strains, 10 to both groups, and 5 to cephalosporins alone. Similar strains were isolated from the inhabitants of the aquaculture surrounding areas. Irrespective of source of samples, strains resistant to all examined antibiotics, carried predominantly the carbapenemase gene blaKPC either alone or with the ß-lactamase genes (blaCTX-M-15, blaSHV, blaTEM, and blaPER-1). The isolates from fish, water, and workers harboured a wide-range of multi-drug-resistance Inc. plasmids, which were similar among all isolates. CONCLUSION: The present findings suggest transmission of the resistance genes among Enterobacteriaceae strains from different sources. This reiterates the need for control strategies that focus on humans, animals, water, and sewage systems to solve the antibiotic resistance problem.

Virulence and Antibiotic Resistance Patterns of Extended-Spectrum Beta-Lactamase-Producing Salmonella enterica serovar Heidelberg Isolated from Broiler Chickens and Poultry Workers: A Potential Hazard.

The current study investigated the emergence of multidrug-resistance (MDR), extended-spectrum beta-lactamase (ESBL)-producing Salmonella enterica serovar Heidelberg in broiler chickens and workers in poultry farms. A total of 33 S. Heidelberg isolates were recovered; 24 from the broiler cloacal swabs and 9 from the farm workers. All the S. Heidelberg isolates were tested for susceptibility to 11 antimicrobial agents and for the presence of resistance and virulence genes. MDR strains were found in 95.8% (23/24) and 88.8% (8/9) of the broiler and human isolates, respectively. Among the MDR strains, 66.6% of the broiler isolates and 55.5% of the human isolates were ESBL producing. The majority of broiler isolates showed resistance to ampicillin (100%) and ceftriaxone (91.6%), followed by ceftazidime and imipenem, (87.5%) and (75%). The resistance rate of the human isolates to those antibiotics were lower than the broiler isolates; ampicillin (88.8%), ceftriaxone (66.6%), ceftazidime (77.7%), and imipenem (66.6%). The resistance determinant genes found among the isolated strains was blaSHV-1, blaTEM-1, blaCMY-2, blaOXA-1, blaCMY-M2, blaPSE-1, and ampC. The most detected ESBL genes for broiler and human isolates were ampC (63.7%) and blaSHV-1 (56.6%), followed by blaCMY-M2 (48.5%), blaTEM-1 (39.4%), and blaOXA-1 (27.3%); whereas blaCMY-2 and blaPSE-1 were not detected. The finding of chromosomal and plasmid virulence genes revealed that the invA (100%), stn, sipC, and rck (72.8%), spvC (66.7%), ssr (63.6%), sopB (54.6%), and hilA and sipA (3.0%), while pefA and ssaR were absent. An elevated rate of MDR Salmonella Heidelberg in chickens is of potential great health risk. This signifies the role of the food of animal origin as a reservoir of MDR Salmonella that can affect the human health.

Antimicrobial action of autologous platelet-rich plasma on MRSA-infected skin wounds in dogs.

Effective antimicrobial preparations, other than antibiotics, are important for the treatment of potentially fatal drug-resistant infections. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of hospital-acquired and post- operative infections. Fortunately, the antimicrobial properties of platelet-rich plasma (PRP) against various microorganisms enable its potential use as an alternative to conventional antibiotics. The present work was designed to evaluate the hypothesized antimicrobial activity of PRP against MRSA infected skin wounds. Six adult male dogs were divided equally into control and PRP groups. Unilateral circular full-thickness skin wounds were created then a MRSA suspension was injected locally. Treatment started at 1st week post infection with subcutaneous infiltration of autologous activated PRP every week in the PRP group and with topical application of clindamycin cream twice daily in the control group. PRP decreased wound size and significantly increased wound contractility and re-epithelization, as confirmed by histopathological and immunohistochemical findings. Also PRP treated group showed significant decrease in ROS and redox imbalance with over expression of the TNF-α and VEGFA genes that indicate angiogenesis and maximum antibacterial activity after three weeks. In conclusion, CaCl2-activated PRP exhibited antimicrobial activity against MRSA infection, which improved the infected wound healing re-epithelization and granulation tissue formation.

Vancomycin-resistant Staphylococcus aureus isolated from camel meat and slaughterhouse workers in Egypt.

Background: The emergence of vancomycin-resistant Staphylococcus aureus (VRSA) represents a challenge for the treatment of staphylococcal infections in both human and animals worldwide. Although VRSA has been detected in several animal species worldwide, data on the bacterial prevalence in dromedary camels and workers in camel slaughterhouses are scarce. Methods: We investigated meat samples from 200 dromedary camel carcasses from three different abattoirs that were being prepared to be sent to the markets. Twenty hand swabs were voluntarily collected from the workers in the same abattoirs. Isolation and identification of the bacterial specimens from the samples were performed using conventional cultural techniques and biochemical identification and were confirmed by PCR amplification of the nuc gene. Antimicrobial susceptibility against nine antimicrobial agents commonly used in human and camels was tested using the disc diffusion method, and genetic analysis was performed by evaluating the mecA gene in phenotypically oxacillin (OXA)- and cefoxitin (FOX)-resistant isolates. The resistance of S. aureus to vancomycin (VAN) was tested by broth microdilution and confirmed by PCR targeting the vanA and vanB genes. The vanA and vanB genes were sequenced. Result: S. aureus was detected in both camel meat (29/200, 14.5%) and in abattoir workers (11/20, 55%). Of the collected samples, 27% (8/29, camel) and 54% (6/11, human) were identified as VRSA.All VRSA isolates carried both the vanA and vanB genes. Additionally, all VRSA isolates were also classified as methicillin-resistant S. aureus (MRSA). The vanA amplicons of the isolates from human and camel meat were homologous and clustered with a Chinese reference isolate sequence. Conclusion: This study demonstrated that VRSA is present in camel abattoirs in Egypt. Zoonotic transmission between animals and human is probable and reflects both a public health threat and a food safety concern.

Occurrence of cagA+ vacA s1a m1 i1 Helicobacter pylori in farm animals in Egypt and ability to survive in experimentally contaminated UHT milk.

Cases of human gastric cancer due to Helicobacter pylori have been reported worldwide and animals might act as a reservoir of infection in certain circumstances. The recent few decades showed a rapid decline in the incidence of gastric cancer, which was mainly due to the decrease in H. pylori infection. The aims of the present study were to determine the prevalence of H. pylori among livestock and investigate whether the animal isolates can be transmitted through contaminated milk causing gastric infection. Feces and milk samples were collected from apparently healthy cows, buffaloes, and sheep, and were examined by nested PCR and genotyping. The PCR positive samples were further subjected to bacterial culture followed by partial 16s sequencing of the isolates. Twenty-nine percent of the animals showed the presence of H. pylori, mainly the virulent cagA+vacA+s1a m1 i1 genotype, which is known to be associated with serious diseases in humans. The spiral viable culturable form (SVCF) of this strain was inoculated into UHT (ultra-high temperature) milk and remained viable for up to 10 days at 4 °C. Increasing period of storage and or temperature led to a decrease in the number of the SVCF and occurrence of the coccoid viable non-culturable form (CVNCF). The infectivity of the survived forms was determined by feeding healthy groups of laboratory mice with the contaminated UHT milk containing SVCF or CVNCF for 40 days. The gastric mucosa of the two mice groups showed similar levels of H. pylori load. This highlights that H. pylori can persist in contaminated milk by entering a non-culturable state, which can induce gastric infection.

Genotyping and antimicrobial resistance patterns of Helicobacter pylori in human and dogs associated with A2142G and A2143G point mutations in clarithromycin resistance.

BACKGROUND: Routes of transmission of Helicobacter pylori a class I carcinogen bacterium and the roles of animals have not been yet well determined. This study was carried out to investigate H. pylori phenotypically and genotypically in human and dogs to determine the antibiotic resistance patterns. As eradication therapy depends mainly on clarithromycin we evaluated 23S rRNA gene mutations associated with its resistance. RESULTS: A total of 150 human stool samples and 60 canine gastric biopsies were examined by nested PCR for the presence of H. pylori, 60% and 76.6% were positive respectively. Only 20 (22.2%) and 41 (89.1%) isolates were successfully cultured from human and canine samples respectively. Genotyping revealed a total of cagA+vacA+ combinations 76.6% (69/90) and 65.2% (30/46) in human and dogs, respectively. Allelic diversity in vacA gene was obviously observed, while cagA-vacA+ combinations were 23.3% (21/90) and 34.7% (16/46) in human and dogs, respectively. The antimicrobial susceptibility patterns of human exhibited the highest levels of resistance against Clarithromycin (60%), Trimethoprim (55%), metronidazole (45%), amoxicillin (45%) and cefsulodin (60%) antibiotics and comparatively lower for spiramycin (10%) and tetracycline (15%). Dogs strains showed the highest levels of resistance against Clarithromycin (53.6%), metronidazole (51.2%) and erythromycin (43.9%) antibiotics, on the other hand, the percent of resistant canine strains were comparatively lower for spiramycin (9.7%). Single point mutation of A2143G was detected as 25% (3/12), 18.1% (4/22) in human and dogs respectively. Single point mutation of A2142G was detected as 16.6% (2/12), 13.6% (3/22) in human and dogs, respectively. While dual mutations of both A2142G and A2143G were detected as 50% (6/12), 40.9% (9/22) in human and dogs, respectively. CONCLUSION: occurrence of elevated rates of A2142G and A2143G point mutations in clarithromycin resistant H. pylori isolates from human and dogs causing failure in treatment and eradication of the pathogen. The roles of animals need attention and further investigations.

New Insight of Apparently Healthy Animals as a Potential Reservoir for Clostridium Perfringens: A Public Health Implication.

INTRODUCTION: Clostridium perfringens is commonly found in the gastrointestinal tract of animals and humans and continues to cause one of the most prevalent foodborne diseases in man. MATERIAL AND METHODS: A total of 355 samples were examined for the occurrence of C. perfringens: rectal swabs from cattle, sheep, and goats, fresh stool samples from diarrhoea sufferers having been in contact with these animals, irrigation water and soil samples from the husbandry sites, and pre-harvesting fresh produce from farms irrigated with the sampled water. All samples were collected from Cairo and Giza governorates, Egypt. PCR analysis was carried out with positive isolates using the α-toxin gene. Sequence analysis of the gene of C. perfringens isolates was performed using the neighbour-joining approach. Bootstrap analysis was executed with 1,000 resamplings. RESULTS: 174 C. perfringens strains were isolated with a 49.01% prevalence. The highest prevalence of C. perfringens in apparently healthy animals was found in sheep (65.45%) followed by goats (58%), buffaloes (55%), and cattle (47.1%). Its prevalence in humans being in contact with these animals was 47.5%. The bacterium's isolation from the soil and irrigation water was achieved in 40% and 31.7% of samples, respectively, posing a risk, particularly when the water and soil contact food in the field, shown by the fresh produce isolation of 40%. A significant relationship between the prevalence of C. perfringens in animal and environmental samples was identified (P < 0.05). A significant relationship was identified neither between animal species and C. perfringens prevalence, nor between the environmental source and C. perfringens prevalence (P > 0.05). All isolates were positive for the α-toxin gene by PCR. The sequence analysis and the phylogenetic relationship of the α-toxin genes from different samples revealed that C. perfringens from faeces of apparently healthy cattle, buffaloes, sheep, and goats is a significant threat in places where it can contaminate the soil and water. In addition, the sequence of C. perfringens from humans suffering from diarrhoea was found in the same cluster with the sequence from cows, goats, and sheep. CONCLUSION: The role of apparently healthy animals in transmitting C. perfringens to humans, either through being in direct or indirect contact via water or soil in the cultivation of vegetables and fruits, was demonstrated.

Extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa in camel in Egypt: potential human hazard.

BACKGROUND: The rapid increase of extended-spectrum beta-lactamase (ESBL) producing bacteria are a potential health hazard. Development of antimicrobial resistance in animal pathogens has serious implications for human health, especially when such strains could be transmitted to human. In this study, the antimicrobial resistance due to ESBL producing Pseudomonas aeruginosa in the camel meat was investigated. METHODS: In this study meat samples from 200 healthy camels at two major abattoirs in Egypt (Cairo and Giza) were collected. Following culture on cetrimide agar, suspected P. aeruginosa colonies were confirmed with a Vitek 2 system (bioMe´rieux). P. aeruginosa isolates were phenotypically identified as ESBL by double disk synergy test. Additionally antimicrobial susceptibility testing of ESBL producing P. aeruginosa isolates were done against 11 antimicrobial drugs and carried out by disk diffusion method. The ESBL genotypes were determined by polymerase chain reaction according to the presence of the bla PER-1, bla CTX-M, bla SHV, and bla TEM. RESULTS: Pseudomonas aeruginosa was isolated from 45 camel meat sample (22.5%). The total percentage of ESBL producing P. aeruginosa was 45% (21/45) from camel meat isolates. Antibiogram results revealed the highest resistance was for c, ceftriaxone and rifampicin followed by cefepime and aztreonam. The prevalence rates of ß-lactamase genes were recorded (bla PER-1 28.5%, bla CTX-M 38%, bla SHV 33.3% and bla TEM 23.8%). CONCLUSIONS: This study illustrates the presence of high rates of ESBL-P. aeruginosa in camels that represents an increasing alarming for the risk of transmission to human and opens the door for current and future antibiotics therapy failure. Livestock associated ESBL-P. aeruginosa is a growing disaster, therefore, attention has to be fully given to livestock associated ESBL-bacteria which try to find its way to human beings.

Serologic evidence and risk factors for Helicobacter pylori infection in animals and humans.

INTRODUCTION: Helicobacter pylori (H. pylori) is one of the most common bacterial infections among humans worldwide. Although many records imply its interfamilial acquisition, the role of animals remains poorly understood. This study was undertaken to investigate the seroprevalence of H. pylori in animals and their human contacts in Cairo and Giza governorates, Egypt. METHODOLOGY: Commercial enzyme-linked immunosorbent assay (ELISA) kits were used to detect IgG antibodies to H. pylori in dogs, cattle, and humans. RESULTS: Seropositive dogs (35/94; 37.2%), cattle (24/80; 30%) and humans (40/90; 44.4%) were found. Seroprevalence in animals significantly varied in different areas of sample collection, but there was no association with sex or age. Human seropositivity rates were associated with increasing age; moreover, seropositive dog owners (51.7%; 15/29), had seropositive dogs. However, infection was not associated with subject's sex, occupation, or history of animal contact. CONCLUSIONS: Our findings indicate H. pylori is widely distributed in cattle and dogs and their human contacts in Cairo and Giza, Egypt. Further studies to determine infection in other occupational groups are needed. This study provides baseline information on the seroprevalence of H. pylori, which may be required to begin prevention control programs in our area.

Antimicrobial Resistance, Biofilm Formation and mecA Characterization of Methicillin-Susceptible S. aureus and Non-S. aureus of Beef Meat Origin in Egypt.

Methicillin-resistant Staphylococcus aureus (MRSA) have been found in various farm animal species throughout the world. Yet, methicillin-susceptible S. aureus (MSSA), methicillin-susceptible non-S. aureus (MS-NSA), and methicillin-resistant non-S. aureus (MR-NSA) were not investigated. Therefore, we persued to determine the diversity in their phenotypic virulence assay, phenotypic antimicrobial resistance profile and molecular characterization in one of the food chains in Egypt. Samples were collected during 2013 from beef meat at retail. Twenty seven isolates comprising five species (S. hyicus, S. aureus, S. schleiferi subsp. coagulans, S. intermedius, and S. lentus) were characterized for their antibiotic resistance phenotypic profile and antibiotic resistance genes (mecA, cfr, gyrA, gyrB, and grlA). Out of the 27 Staphylococcus isolates only one isolate was resistant to the 12 antibiotics representing nine classes. Raw beef meat sold across the Great Cairo zone, contains 66.7% of MRS, with highest prevalence was reported in S. aureus (66.7%), while the MRS non-S. aureus strains constituted 66.7% from which S. hyicus (60%), S. intermedius (33.3%), S. schleiferi subsp. coagulans (100%), and S. lentus (100%) were MRS. Seven S. aureus, six S. hyicus, four S. schleiferi subsp. coagulans, three S. intermedius, and one S. lentus isolates although being resistant to oxacillin yet, 11/27 (40.7%) carried the mecA gene. At the same time, the cfr gene was present in 2 of the nine S. aureus isolates, and totally undetectable in S. hyicus, S. schleiferi subsp. coagulans, S. intermedius, and S. lentus. Although, global researches largely focused into MRSA and MR-NSA in animals on pigs, the analysis of our results stipulates, that buffaloes and cattle could be MRSA dispersers and that this theme is not specific to pigs. Detection of MSSA virulence determinants is a must, as although oxacillin resistance may be absent yet, the MSSA may carry the virulence determinants which could be a source of perilous S. aureus for the human community.

Eucalyptus Tree: A Potential Source of Cryptococcus neoformans in Egyptian Environment.

In Egypt, the River Red Gum (Eucalyptus camaldulensis) is a well-known tree and is highly appreciated by the rural and urban dwellers. The role of Eucalyptus trees in the ecology of Cryptococcus neoformans is documented worldwide. The aim of this survey was to show the prevalence of C. neoformans during the flowering season of E. camaldulensis at the Delta region in Egypt. Three hundred and eleven samples out of two hundred Eucalyptus trees, including leaves, flowers, and woody trunks, were collected from four governorates in the Delta region. Thirteen isolates of C. neoformans were recovered from Eucalyptus tree samples (4.2%). Molecular identification of C. neoformans was done by capsular gene specific primer CAP64 and serotype identification was done depending on LAC1 gene. This study represents an update on the ecology of C. neoformans associated with Eucalyptus tree in Egyptian environment.

Lyme borreliosis: A neglected zoonosis in Egypt.

Borrelia burgdorferi, the causal organism of Lyme borreliosis. In Egypt, available data about the occurrence of Lyme disease are scarce and no structured studies documented the presence of Lyme borreliosis in Egyptian animals and tick reservoirs verifying its zoonotic evidence. Besides, no successful trials to isolate B. burgdorferi from clinical samples have occurred. This study was conducted to investigate B. burgdorferi infection as an emerging zoonosis neglected in Egypt. A total number of 92 animals, tick and human companion specimens were collected and subjected for culture, PCR and/or serodetection. B. burgdorferi has been detected and isolated from Egyptian animal breeds. We also detected the presence of outer surface protein A gene of B. burgdorferi by PCR as well as anti-B. burgdorferi IgM by ELISA in human contacts who were suffering from fever of unknown origin. This report represents the first systematic study on animals associated with patients suffering from febrile illness to confirm the emerging of such neglected zoonosis in Egypt.