Background and Aim: Mycobacterium tuberculosis complex (MTBC) is a group of mycobacteria that are important human pathogens. Mycobacterium tuberculosis and Mycobacterium bovis cause serious chronic life-threatening disease and also significant economic losses in both production and remedication. Recently, emergence of multidrug-resistant tuberculosis (MDR-TB) complex has generated global recognition of the need for rapid and sensitive diagnosis and development of new treatments. The current study illustrates the isolation/identification of MTBC strains in specimens obtained from cows and humans by conventional and real-time polymerase chain reaction (RT-PCR) techniques. Further, the study assesses sensitivity to antituberculosis drugs in isolated MDR strains. Materials and Methods: A total of 1464 samples from cattle (1285 raw milk and 179 lymph node), and 149 human sputum samples, were collected from farms and abattoirs in Delta Egypt. Conventional methods (culture and Ziehl–Neelsen staining) were implemented as were RT-PCR using MTBC universal DNA. The effect of some antituberculosis drugs on obtained isolates was assayed using drug susceptibility proportion and qualitative suspension techniques. Results: The MBTC detection rate using the culture method was higher than for Ziehl–Neelsen staining; raw cow milk (2.56 vs. 1.63%), lymph nodes (51.59 vs. 48.04%), and human sputum (5.36 vs. 4.02%). A total of 135 isolates were obtained. Application of RT-PCR detected 138 isolates from the same set of samples. MBTC isolates were resistant to first-line antituberculosis drugs, such as pyrazinamide, isoniazid, rifampicin, and ethambutol by 78.5, 59.3, 40.7, and 31.8%, respectively, and could be highly resistant to kanamycin (82.3%) and amikacin (80.7%). However, isolates remained sensitive to ciprofloxacin (71.1%) and clarithromycin (73.3%) as second-line drugs. Conclusion: There is a growing risk for isolation of MDR-TB from raw milk and lymph nodes of field tuberculin positive cattle as well as sputum of veterinarians and workers existed in farms and abattoirs. PCR-based techniques have become the gold standard for the identification of mycobacterial species, showing high efficiency compared to bacteriological and microscopic examination. Application of the first- and second-line antituberculosis drugs in combination could counter the MDR-TB concern once infections are identified.

Objective: Animal trade has an important role in the economy but in contrast, it causes the spread of infectious diseases overall the world, in particular, the trans-boundary animal diseases. Therefore, the aim of this study is to report the prevalence rate of some transboundary infectious diseases to assess the effectiveness of quarantine measure in the detection of exotic disease and clarify the role of live animal trade in infectious transboundary diseases spread. Materials and Methods: The study was done on 176 serum samples obtained from cattle imported from Sudan in order to determine the prevalence of foot and mouth disease (FMD), Peste Des Petits Ruminants (PPR), and Infectious Bovine Rhinotracheitis (IBR). Three serological tests were used; Serum neutralization test for FMD, Indirect enzyme-linked immunosorbent assay (i-ELISA) for PPR, and Competitive ELISA for IBR. Results: The seroprevalence of FMD in tested sera was; 77.27% in the serotype A (A-Iran), 68.18% in the serotype A (A-Africa), 93.82% in the serotype O (O-Pan Asia), and 35.227% in the serotype South African Territories-2 (SAT-2) SAT-2. While the overall seroprevalence of PPR was 49.431% and the IBR was 93.75%. Conclusion: The result indicates the serious role of live animal trade as “hubs” for infectious diseases spread. Subsequently, the common control measures must be taken to avoid the spread of the diseases through the animal trade; which include screening, surveillance, precautions at borders, and vaccination.

Echinococcus spp. are important cosmopolitan zoonotic parasitic tapeworms that cause a disease called hydatidosis or cystic echinococcosis (CE), which has remarkable economic losses. The objective of our study was to develop a specific IgG polyclonal antigen-based ELISA (Sandwich ELISA; capture ELISA) method for the detection of circulating Echinococcus granulosus (E. granulosus) antigens in camels infected with hydatid cysts before slaughtering and its application in serodiagnosis of CE in animals to assess the positive rate of hydatidosis in camels slaughtered in Giza governorate abattoirs in Egypt. In this study, molecular identification of Echinococcus sp. isolate was performed based on the NADH dehydrogenase subunit 1 (NAD1) gene, revealing the isolate (GenBank: OQ443068.1), which is identical to the G6 E. granulosus sensu lato genotype. The positive rate of hydatid cysts was determined in slaughtered camels’ organs (n = 587). The results revealed that hydatid cysts were found in 46.5% (273/587) of the examined camels. Pulmonary echinococcosis was significantly more prevalent in the slaughtered camels (60%, 164/273) than hepatic echinococcosis (39.9%, 109/273), (p = 0.001, Chi Square = 11.081). Cyst fertility rates were higher in hepatic (90.8%, 99/109) than in pulmonary cysts (83.5%, 137/164) and the most viable protoscoleces were recorded from fertile the hepatic cysts (67.85 ± 12.78). In this study, hydatid cyst germinal layer antigen (GlAg) was isolated and used for the immunization of rabbits to raise IgG polyclonal antibodies (anti-Echinococcus GlAb IgG). These IgG polyclonal antibodies were purified by affinity chromatography using a protein A column, then labeled with horseradish peroxidase. Electrophoretic analysis of IgG polyclonal antibodies and crude GlAg was performed in 10% polyacrylamide gels. The SDS-PAGE revealed four bands at molecular weights of 77 kDa, 65 kDa, 55 kDa, and 25 kDa. The Sandwich ELISA was performed to evaluate the sensitivity and specificity and cross-reactivity of the prepared IgG polyclonal antibodies. The circulating hydatid antigen was found in 270 out of the 273 samples with hydatidosis, with a sensitivity of 98.9% (270/273), a specificity of 94.9% (296/312) and a diagnostic efficacy of 96.8%. Regarding the cross reactivity, anti-Echinococcus GlAb IgG showed a low cross-reactivity with Fasciola gigantica infected camel sera (3/8), and Myiasis (Cephalopina titillator larvae; 3/20). No cross-reactivity was recorded with uninfected camel sera (negative sera for E. granulosus), and no cross-reactivity was found with antigens of Eimeria spp., Toxoplasma gondii, Cryptosporidium sp., and Hyalomma dromedarii (ticks’ infestation). Then, Sandwich ELISA was conducted again to detect E. granulosus antigen in all the collected camel sera, which resulted in a 48.7% (286/587) positive rate of CE compared to 46.5% (273/587) using a postmortem inspection (PM diagnosis) (p = 0.5, Chi Square = 0.302). In conclusion, the Sandwich ELISA technique introduced in this study appears to be a sufficiently sensitive diagnostic assay for the detection of camels’ echinococcosis using anti-Echinococcus GlAb IgG. In addition, it might offer a significant medical and veterinary importance in helping the early detection of hydatidosis, as well as its early treatment.

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The current study was applied to investigate E. coli and Staph. aureus counts as well as serodiagnosis of E. coli, Salmonellae isolates and determination of enterotoxin production by Staph. aureus in 100 random local and imported fresh beef samples, from different slaughterhouses at Cairo and Giza Governorates. Staph. aureus was isolated from 22(44%) and 20 (40%) of examined local and imported meat samples respectively. E. coli was found in 40 (80%) and 34(68%) in examined local and imported meat samples, respectively. Staph. aureus enterotoxin types (A & D) were detected in two strains out of 5 randomly selected isolates from local meat only (20% of each). E. coli, one strain (20%) was isolated from fresh local meat; and classified as Enteropathogenic (EPEC), while two strains (40%) were Enterohaemorrhagic (EHEC) "O111:H2", one strain (20%) was Enterotoxigenic (ETEC) "O15:H4". On the other hand, one strain of E. coli (20%) isolated from imported meat was classified as Enteropathogenic (EPEC) and its serodiagnosis was O128:H2, while one strains (20%) was Enterohaemorrhagic (EHEC) "O26:H11" which harbored both (stx1 and stx2) genes and finally, the 3 rd strain (20%) was Enteroinvasive (EIEC) "O159". Meanwhile, Salmonella spp. was isolated from two samples (4%) of examined local meat; one strain was identified as S.

This study was conducted to evaluate the bacteriological safety of raw and cooked meat products represented by Aerobic Plate Count, Enterobacteriaceae, Coliforms, E. coli and Staph. aureus counts as well as determination of pH, TVBN and TBA of a totally examined 110 samples (20 each of shish tawook, kofta, fillet and escalope pane, besides, 30 swabs (10 each of hands, food contact surfaces and cutting knifes). The obtained results in this study indicated that the Overall reduction the bacterial counts of the above-mentioned organisms according to their prevalence in the different products under study were significantly reduced by 2, <1, 1, and <2 log10cfu/g in raw examined samples of shish tawook, kofta, fillet and escalope pane treated with 5% vinegar as compared with raw untreated samples, respectively. In addition, cooked treated aforementioned sample types with vinegar 5% showed significant reduction represented by 2, 1, <2 and 1 log10cfu/g when compared with cooked untreated samples. So, using of vinegar either in raw or cooked products leads to a significant reduction in the number of bacteria contaminating products. Hand swab before cleaning and sanitizing recorded 3.31±0.52 (100%), 2.54±0.92 (100%), 2.47±0.74 (60%), <1 (0.0%) and 2.12±0.55 (80%), while after cleaning it was recorded 2.39±0.5 (100%), 1.59±0.25 (60%),1.36±0.24 (40%), <1 (0.0%) and 1.74±0.62 (60%). Surfaces swab recorded 3±1.04 (100%), 1.5±0.96 (50%), 2.06±1.49 (20%), <1 (0.0%) and 2.3±1.18 (30%), before cleaning and 1.73±1.02 (100%), <1, <1, <1 and (30%) 2±0.7 log10cfu/g after cleaning. Knife swabs recorded 3.35±0.76 (100%), 2.68±0.68 (50%), 2.5±0.91 (40%), <1and 2.25±1.0 (70%) before cleaning, while after cleaning they were recorded 1.77±1.19 (100%), 2.59±0.16 (20%), <1, <1and 1.97±0.58 for the aforementioned bacterial counts, respectively. Salmonella was not isolated from any of the examined samples or swabs. Meanwhile, chemical analysis of pH, TVB-N and TBA recorded 0.04±0.02, 11.86±1.24 and 5.86±0.18 for shish tawook; 0.05±0.02, 6.6±2.19 and 6.12±0.31 for kofta; 0.04±0.03, 7.22±2.49 and 5.62±0.13 for fillet and 0.41±0.16, 13.18±2.15 and 5.96±0.49 for escalope pane, respectively. Significance differences (P<0.05) were obvious between raw and cooked samples and for some extent between treated and non-treated one.

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Background and Aim: Brucellosis is a zoonotic disease with a worldwide distribution. It has a serious impact on the health of humans and animals, along with a negative impact on the economy. This study aimed to prepare and evaluate the diagnostic performance of a lateral flow immunochromatographic test (LFIT) nanogold diagnostic kit for detecting brucellosis in sheep. Materials and Methods: A rapidly developed LFIT, in which lipopolysaccharide conjugates with nanogold molecules, was placed on the conjugate pad. One hundred ovine serum samples were tested to detect Brucella antibodies (Ab) using the prepared lateral flow immunochromatography assay (LFA) kit and Rose Bengal test. The evaluation of specificity, sensitivity, and accuracy for LFIT and Rose Bengal plate test was conducted using the P04310-10 IDEXX brucellosis ovine/ caprine Ab enzyme-linked immunosorbent assay (ELISA) test (gold standard). Results: The lower amount of Brucella Ab in the ovine serum samples was detected and was 1.58 S/P ratio ELISA titer/100 μL using LFIT and with Rose Bengal to detect 1.86 S/P ratio ELISA. The results showed that the developed LFIT had high specificity with no cross-reactivity with other tested bacteria. The calculated sensitivity, specificity, and accuracy of LFIT and Rose Bengal test using the P04310-10 IDEXX brucellosis ovine/caprine Ab ELISA test (gold standard) were 74% and 89%, 81% and 59%, and 76.9% and 66%, respectively. Conclusion: The present results showed interesting results implying that the LFIA strip test could be used as a substantial diagnostic tool for field screening ovine Brucella as an essential step in the control of brucellosis. However, further studies for the validation of the present findings are necessary.