Prevalence of intestinal parasites with molecular detection and identification of Giardia duodenalis in fecal samples of mammals, birds and zookeepers at Beni-Suef Zoo, Egypt.

The current study aimed to investigate the prevalence of intestinal parasites from various species of mammals and birds housed in a zoological garden in Beni-Suef province, Egypt. A total of 77 fecal samples were collected from various primates (16), carnivores (7) and herbivores (54). Meanwhile, 123 fecal samples were collected from two Ostrichs (Struthio camelus), five Numida meleagris (Numida meleagris), twoIndian Peafowls (Pavo cristatus), two Emu (Dromaius novaehollandiae) 101 Pigeons (Columba livia domestica) and 11 Swan Goose (Anser sygnoides). In addition, seven stool samples from zookeepers who had been in close contact with animals and birds were examined. Salt flotation and formol ether sedimentation techniques were applied for parasitological examination. Positive samples of Giardia cysts were preserved in alcohol and kept at 4 °C until DNA extraction. Parasitological findings revealed that 48.05% of zoo animals were infected with intestinal parasites; 11.69% were positive with helminths and 27.27% with protozoa, however 9.09% had mixed infection. It was found that 75%, 57.14% and 38.89% of primates, carnivores and herbivores respectively were infected with intestinal parasites. In Primates the most prevalent parasites were Giardia spp. (43.75%) then Entamoeba histolytica/dispar (12.5%), Entamoeba coli (12.5%) and Trichuris spp. (6.25%). For carnivores, Ancylostomatidae had the highest prevalence (42.86%) followed by Spirometra spp. (14.29%). Meanwhile, Eimeria spp. (20.37%) was the most prevalent parasite in herbivores, followed by Blantidium coli (7.41%) and Tricuris spp. (7.41%), then Toxocara spp. (3.70%). Furthermore, the prevalence of infection in zoo birds was 21.95%. The identified parasites were Eimeria spp., Giardia spp., Capillaria spp., Ascaridia spp., Isospora spp. and Sublura brumpti. Stool examination of zookeepers revealed the presence of G. doudenalis and E. histolytica/ dispar cysts. The gdh gene of G. duodenalis was successfully amplified from fecal samples of zoo mammalsand zookeepers. In conclusion, the application of preventive and control measures against the propagation of infectious intestinal parasites is essential to prevent the spread of these parasites among zoo animals or to humans. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12639-020-01341-2.

Efficiency evaluation of some novel disinfectants and anti-bacterial nanocomposite on zoonotic bacterial pathogens in commercial Mallard duck pens for efficient control.

OBJECTIVE: This work aimed to detect the frequency of pathogenic bacteria of zoonotic importance in ducks' dropping, their surrounding environment, and farmworkers in contact with them. Furthermore, the susceptibility pattern of isolated bacteria to antimicrobial drugs and the efficiency of disinfectants (CID 20, Durak® plus, and hydrogen peroxide (H2O2), nano zinc oxide (ZnO NPs), and hydrogen peroxide loaded nano zinc oxide (H2O2/ZnO NPs) composites against isolated bacteria were evaluated. MATERIALS AND METHODS: A total of 271 samples were collected from duck pens, including 35 fecal droppings, 200 environmental samples, and 36 from the hands of pen workers for isolation and identification of bacterial strains using standard microbiological procedures. After that, the antibiotic sensitivity testing of 40 bacterial isolates was carried out using disk diffusion assay. ZnO NPs and H2O2/ZnO NPs were characterized using Fourier-transform infrared spectrum and high-resolution transmission electron microscopy. The efficacy of disinfectants and nanocomposites was evaluated against enteropathogenic bacteria using the broth macro-dilution method. RESULTS: The results showed that the overall prevalence of pathogenic bacteria in duck pens was 62.73. The highest isolation rate was detected in duck fecal droppings (100%), while Escherichia coli was found to be the most isolated pathogen (56.47%), followed by Pseudomonas aeruginosa (21.8%), Proteus mirabilis (15.29), and Salmonella species (6.47%). Multidrug resistance (MDR) was detected in the majority of bacterial isolates. The efficiency of CID 20 and Durak® plus disinfectants against all bacterial isolates was highly susceptible (100%) after 120 min of exposure time compared to the effectiveness of H2O2 on enteropathogenic bacteria which did not exceeded 60% at 5% concentration. Meanwhile, the sensitivity of Salmonella spp. to Durak® plus did not exceeded 80%. CONCLUSION: The duck fecal droppings are the primary source of bacterial isolates. MDR isolates were susceptible to both CID 20 and Durak® plus disinfectants after 120 min of exposure time at a concentration of 1:100 ml. Besides, H2O2/ZnO NPs composite proved its lethal effect against all testing strains at 0.02 mg/ml after 120 min of exposure. Strict biosecurity guidelines are required to mitigate and prevent the transmission of potentially zoonotic pathogens through the farm environment and/or duck droppings.

Prevalence, Characterization, and Control of Campylobacter jejuni Isolated from Raw Milk, Cheese, and Human Stool Samples in Beni-Suef Governorate, Egypt.

Our study aimed to determine the prevalence of Campylobacter jejuni isolated from raw milk, cheese, and human stool samples in Beni-Suef Governorate, Egypt, and to characterize the antibiotic resistance profile and virulence genes of the isolates. An additional objective was to evaluate the effectiveness of cinnamon oil and Lactobacillus acidophilus La5 for controlling C. jejuni in cheese. A total of 200 samples of raw milk and dairy products, including 50 samples of raw milk and 150 samples of three different types of cheese were used. Fifty-three human stool samples were also collected. The samples were tested for the presence of C. jejuni using culture and molecular methods. Campylobacter spp. were isolated from 9.5% (19/200) of the raw milk and cheese samples. The highest prevalence was observed in milk samples (18%), followed by Kareish cheese (14%) and Talaga cheese (6%). In contrast, C. jejuni was not found in any of the Feta cheese samples. Of the human stool samples, 21 (39.6%) were positive for C. jejuni. Of the isolates, 60-90% were highly resistant to the antimicrobial agents tested, that is, nalidixic acid, ciprofloxacin, and tetracycline. Virulent cadF and cdtA genes were detected in all isolates. As milk and dairy products are important sources of contamination, reducing the level of C. jejuni in them will lower the risk to consumers. We showed that L. acidophilus La5 was able to control C. jejuni in Kareish cheese, but cinnamon oil was less effective.

Ecological study on antimicrobial-resistant zoonotic bacteria transmitted by flies in cattle farms.

Flies were qualitatively and quantitatively monitored on both livestock animals and the surrounding environment to investigate their role as a potential carrier for antimicrobial-resistant bacteria of zoonotic importance in cattle farms. This was done by the use of visual observations and animal photography; meanwhile, in the surrounding environment, flies were collected using sticky cards and then microscopically identified. Representative fly samples were cultured for bacterial isolation, biochemical identification, and then tested against common 12 antibiotics. The total average of dipterous flies in examined farms was 400.42 ± 6.2. Culicoides biting midges were the most common existing species (70.01 %) followed by house flies, stable flies, and mosquitoes (18.31, 7.74, and 3.91 %, respectively) at X (2) = 9.0, P < 0.05. The most predominant bacterial isolates were Escherichia coli (22.6 %), Staphylococcus aureus and Enterobacter (17.3 % each), coagulase-negative Staphylococci (CNS) (14.7 %), Klebsiella sp. (8 %), Salmonella spp. (6.7 %), and Shigella spp. and Proteus spp. (6.7 % each). The tested bacterial isolates were resistant to variant antibiotics used. S. aureus exhibited 100 % resistance to colistine. However, E. coli revealed 92.9 and 78.6 % resistance against tetracycline and colistine, respectively. Both Salmonella spp. and Shigella spp. were 100 % resistant to penicillin, and Klebsiella sp. had 100 % resistance to tetracycline. In conclusion, Culicoides biting midges and house flies could be considered as a potential carrier for multi-drug-resistant bacteria of zoonotic importance. Furthermore, cows' environment has an essential role in propagation and wide spread of antimicrobial-resistant bacterial pathogens.

The Use of Multiplex PCR to Determine the Prevalence of Enterotoxigenic Staphylococcus aureus Isolated from Raw Milk, Feta Cheese, and Hand Swabs.

Staphylococcus aureus (S. aureus) can cause mastitis in cattle and, therefore, can be present in milk. This study was undertaken to determine the prevalence of coagulase positive S. aureus and its enterotoxin genes sea, seb, and sec in isolates recovered from raw milk, feta cheese, and human hand swabs of milk and cheese handlers in Beni-Suef province, Egypt. A total of 100 samples of raw milk and 50 samples of pasteurized-milk feta cheese were collected. In addition, 50 hand swabs from milk handlers and 25 hand swabs from cheese handlers were examined for the presence of coagulase positive S. aureus. The isolates were characterized by multiplex PCR for detection of sea, seb, and sec genes, and for resistance to 5 classes of commonly used antibiotics. Twelve (12/100), 12 (6/50), and 17% (13/75) of milk, cheese, and hand swab samples, respectively, were positive for coagulase positive S. aureus. One isolate was obtained from each positive sample (31 isolates), and none contained genes for SEA or SEC production. Twenty-five percent, 33%, and 31%, respectively, of the isolates contained the genes for SEB, resulting in 3%, 4%, and 5% of samples being positive for toxin producing coagulase positive S. aureus, respectively. At least one isolate was resistant to each of the antibiotics tested. Despite the low potential for SEB production shown, preventative measures, such as maintenance of the cold-chain and good hygienic practices should be implemented to further reduce the potential risk to public health from SEB, and to reduce the spread of antimicrobial resistance.