Some Studies on mycoplasmosis infection in small ruminants using modern diagnostic methods. , Mohamed, Zeinab R., El-Shafey Dina Y., Abdou Nadra-Elwgoud M. I., and ED-Hamed Fadia Abd , Veterinary Medical Journal, Volume 50, Issue 1, p.49-70, (2010)
Infectious Causes of Neonatal Diarrhea in Cattle in Kuwait with Special Reference to Cryptosporidiosis, Majeed, Qais A. H., Al-Batel Maha K., Abdou Nadra-Elwgoud M. I., El-Azazy Osama M. E., Sami Attia M., and El-Said Hassan , Journal of Animal and Veterinary Advances, Volume 10, Issue 17, p.2282 - 2286, (2011) infectious_causes_-neonatal_diarrhea-cattle-kuwait-cryptosporidiosis.pdf
Evaluation of immunochromatography test for detection of four enteropathogens in the feces of sheep and goats in Kuwait., AlAzemi, Maha S., Majeed Qais A. H., Samy Attia, Henedi Adawia A., Youssef Wessam, and Abdou Nadra-Elwgoud M. I. , Open veterinary journal, Volume 11, Issue 3, p.500-507, (2021) Abstract

Background: Diarrhea in newborn small ruminants continues to be the cause of significant financial loss in sheep and goat farms worldwide. Commercial immunochromatographic (IC) assays have been designed and evaluated to be used for the diagnosis of diarrhea in cattle; however, there are no trials to use rapid tests in small ruminants.

Aim: This study was carried out in Kuwait to evaluate the performance of the rapid immunochromatography test (BoviD-4, BioNote, Inc, Korea) for diagnostics of , rotavirus A (RVA), bovine coronavirus (BCoV), and K99 ( K99) in fecal samples of sheep and goats.

Methods: A total of 85 samples were examined using BoviD-4, and the results were compared with that of polymerase chain reaction for , RVA, and BCoV, whereas for K99 it was by isolation and identification as reference tests.

Results: The kappa test agreement results between the BoviD-4 and reference tests were 0.870 (perfect), 0.783 (substantial), 0.728 (substantial), and 0.281 (fair) for the detection of K99, , RVA, and BCoV, respectively. The sensitivity of BoviD-4 kit was 91.2%, 80.0%, 90.0%, and 37.5% and the specificity was 88.2%, 96.0%, 96.4%, and 92.2% for , RVA, K99, and BCoV, respectively.

Conclusion: The Bovid-4 kit can be used as a rapid pen-side test for spp., K99, and RVA in the field. Nonetheless, care must be taken while interpreting the BCoV results of the kit.

Cross-sectional study and genotyping of rotavirus-A infections in ruminants in Kuwait., Abdou, Nadra-Elwgoud M. I., Majeed Qais A. H., Saad Ashraf A., Mijatovic-Rustempasic Slavica, Bowen Michael D., and Samy Attia , BMC veterinary research, Volume 17, Issue 1, p.245, (2021) Abstract

BACKGROUND: Group A rotaviruses (RVA) are zoonotic pathogens responsible for acute enteritis in human and neonatal ruminants. This research aimed to determine the prevalence of RVA in ruminants (cattle, sheep, and goats) and investigate the circulating RVA genotypes in these animals in Kuwait. We conducted a cross-sectional study to detect RVA in ruminants, using an immunochromatography test (IC), direct sandwich ELISA test, and real-time RT-PCR (RT-qPCR) assay using fecal samples.

RESULTS: A total of 400 cattle, 334 sheep, and 222 goats were examined. The prevalence of RVA was 5.3, 1.2, and 2.3%, respectively, using IC. The ELISA test detected RVA from 4.3% of cattle, 0.9% of sheep, and 1.8% of goats. There was a significant association between the occurrence of diarrhea and the presence of RVA in bovine fecal samples (p-value = 0.0022), while no statistical association between diarrhea and the presence of RVA in fecal samples of sheep and goats was observed (p-value = 0.7250; p-value = 0.4499, respectively). Twenty-three of the IC-positive samples (17 from cattle, two from sheep, and four from goats) were tested using a RT-qPCR RVA detection assay targeting the NSP3 gene. The results showed that 21 of 23 IC-positive samples tested positive by RT-qPCR. Detection of RVA genotypes revealed that G10P[11] was the predominant strain in cattle (58.8%), followed by G8P[1] (11.7%). One sheep sample was genotyped as G8P[1]. In addition, G6P[1] and G6P[14] were detected in goat samples.

CONCLUSION: The present study revealed that the IC was more sensitive in detecting RVA antigen in fecal samples than the ELISA test. A higher occurrence of RVA infection was observed in cattle than in sheep and goats. This study suggests that RVA might be a risk factor of diarrhea in bovine calves less than 2 weeks old. This research also demonstrates the circulation of RVA in sheep and goat populations in Kuwait. Finally, the G10P[11] RVA genotype was the most prevalent genotype identified from cattle samples.

Risk factors of diarrhea in small ruminants in Kuwait., Abdou, N. - E. M. I., Majeed Q. A. H., El-Azazy O. M. E., Tahrani L. M. A., AlAzemi M. S., and Alajmi A. , Iranian journal of veterinary research, Volume 22, Issue 2, p.146-149, (2021) Abstract

BACKGROUND: Diarrhea is the most severe disease affecting small ruminant farms, even though there is continuous progress in the control and prevention strategies.

AIMS: This study aimed to identify some of the risk factors associated with diarrhea in small ruminants in Kuwait.

METHODS: Faecal samples from 556 animals (334 sheep and 222 goats) were collected and examined for different enteropathogens. The collected data from animals and herd management were finally analysed.

RESULTS: Statistical analysis identified that age (pre-weaned; P=0.022, OR=2.38), herd size (large; P=0.037, OR=1.7), and season (dry; P=0.022, OR=2.39) were as non-infectious risk factors of the diarrhea occurrence. It was found that K99 (P=0.000, OR=7.7), spp. (P=0.000, OR=586), and spp. (P=0.000, OR=2.97) were reported more frequently in diarrheic animals than non-diarrheic ones.

CONCLUSION: Pre-weaned small ruminants reared in the large herd were more susceptible to diarrhea. K99, spp., and spp. were the most predominant enteropathogens causing diarrhea in small ruminants in Kuwait. Diarrhea was more common in dry season than in wet season.

Molecular characterization of pathogenic Escherichia coli isolated from diarrheic and in-contact cattle and buffalo calves., Awad, Walid S., EL-SAYED AMR A., Mohammed Faten F., Bakry Noha M., Abdou Nadra-Elwgoud M. I., and Kamel Mohamed S. , Tropical animal health and production, Volume 52, Issue 6, p.3173-3185, (2020) Abstract

Escherichia coli field isolates from calves were characterized and categorized into the most significant diarrheagenic pathotypes using polymerase chain reaction (PCR) assays with different specific primers. The used PCR systems were designed to detect sequences representing the group-specific virulence genes encoding fimbriae f5 (K99), Shiga toxins (stx and stx), heat-stable enterotoxins (st), heat-labile enterotoxins (lt), intimin (eae), hemolysin (hylA), and EAEC heat-stable enterotoxin (astA). In the present work, a total of 150 E. coli field isolates were recovered from 150 fecal swabs collected from 100 diarrheic and 50 apparently healthy in-contact cattle and buffalo calves under 3 months old. Out of these 150 isolated E. coli, 106 isolates from 77 diarrheic and 29 in-contact calves harbored one or more of the investigated virulence genes. The pathotyping of the isolates could classify them into shigatoxigenic E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), and enteroaggregative E. coli (EAEC) with a 30.7, 2.7, 12.7, and 7.3% distribution, respectively. Meanwhile, the detection rates of f5, stx, stx, st, lt, eae, hylA, and astA genes were 17.3, 27.3, 6.7, 10, 37.3, 17.7, 9.3, and 20.7%, respectively. These virulence genes were found either single or in different combinations, such as stx/eae, stx/st/f5, eae/st/f5, or st/lt/f5. Four attaching-effacing shigatoxigenic E. coli isolates (AE-STEC) harboring stx/eae were retrieved from diarrheic calves. Although none of the stx-or eae-positive isolates was verified as O157:H7, STEC isolates detected in apparently healthy calves have potential pathogenicity to humans highlighting their zoonotic importance as reservoirs. Atypical combinations of ETEC/STEC and ETEC/EPEC were also detected in percentages of 14.7 and 2.7%, respectively. Most of these atypical combinations were found more in buffalo calves than in cattle calves. While STEC and EPEC isolates were detected more in cattle calves than in buffalo calves, ETEC isolates were the same in the two species. The pathogenic E. coli infection in calves was recorded to be higher in the first weeks of life with the largest numbers of virulence factor-positive isolates detected at the age of 4 weeks. Histopathological examination of five intestinal samples collected from four dead buffalo calves revealed typical attaching and effacing (AE) lesion which was correlated with the presence of intimin encoding virulence gene (eae). Other lesions characterized by hemorrhagic enteritis, shortening and fusion of intestinal villi and desquamation of the lining epithelium of intestinal mucosa had also been detected.

Epidemiological observations on cryptosporidiosis and molecular characterization of Cryptosporidium spp. in sheep and goats in Kuwait., Majeed, Qais A. H., El-Azazy Osama M. E., Abdou Nadra-Elwgoud M. I., Al-Aal Zein A., El-Kabbany Amira I., Tahrani Laila M. A., AlAzemi Maha S., Wang Yuanfei, Feng Yaoyu, and Xiao Lihua , Parasitology research, Volume 117, Issue 5, p.1631-1636, (2018) Abstract

Molecular epidemiological analysis of cryptosporidiosis in Middle Eastern countries suggests that small ruminants could play a major role in the transmission of Cryptosporidium spp. to humans, with a dominance of Cryptosporidium parvum, especially its IId subtypes. However, little information is available on the epidemiology and risk factors of cryptosporidiosis as well the distribution of Cryptosporidium species/genotypes and subtypes in small ruminants in this area, including Kuwait. In the present study, 47 farms from several areas in Kuwait were visited once during October 2014 to September 2015 to collect data on risk factors associated with Cryptosporidium infection. Fecal samples from 334 sheep and 222 goats were examined for Cryptosporidium oocysts by Ziehl-Neelsen staining (ZN) and antigens by enzymatic immunoassay (EIA). The Cryptosporidium prevalence was higher when samples were examined by EIA than ZN (11.4 and 7.2% in sheep and goats by EIA, compared with 4.2 and 3.6% by ZN, respectively). Young age (less than 3 months) and closed housing system are risk factors of Cryptosporidium infection. A correlation between fecal consistency and the occurrence of Cryptosporidium spp. was observed; non-formed fecal samples were often found positive. Molecular characterization of 30 ovine and caprine samples using PCR-RFLP analysis of the small subunit rRNA gene revealed the presence of C. parvum in 23 samples, Cryptosporidium ubiquitum in five samples, and Cryptosporidium xiaoi in two samples. Sequence analysis of C. parvum at the 60 KDa glycoprotein gene locus identified two subtypes, IIaA15G2R1 and IIdA20G1, with the latter being more common (in 2 and 20 successfully subtyped samples, respectively). Only one subtype of C. ubiquitum (XIIa) was recorded. Cryptosporidiosis in small ruminants apparently poses public health problem in Kuwait.

Molecular biological tools applied for identification of mastitis causing pathogens., El-Sayed, Amr, Awad Walid, Abdou Nadra-Elwgoud M. I., and Vázquez Hugo Castañeda , International journal of veterinary science and medicine, Volume 5, Issue 2, p.89-97, (2017) Abstract

The molecular diagnostic tools became the gold standard of mastitis diagnosis in the last few years. They enable rapid, qualitative, quantitative and large scale diagnosis. In addition to their role in diagnosis, they can identify pathogens at the subspecies level which is necessary for the epidemiological studies. They are increasingly used in mastitis control programs through identification of suitable candidates for vaccine production and through the selection of mastitis resistant cattle breeds. The present molecular techniques are continuously improved and new techniques are developed in order to provide higher sensitivity and specificity and to minimize the costs. The present work aims to provide an overview of the modern molecular tools, discuss why they replaced the traditional tools and became the new gold standard in mastitis diagnosis through comparing both traditional and molecular tools, explore the prospective of the molecular diagnostic techniques in mastitis diagnosis and control and to explore new horizons of using molecular assays in near future.

Molecular diagnosis and matching of rapid and Elisa tests for antigen detection of Rota and Corona viruses in neonatal calves., Saad, Ashraf A., and Abdou Nadra-Elwgoud M. I. , Animal Health Research Journal, Volume 5, Issue 2, p.136-146, (2017)
Cestodes and Nematodes Recorded in Stray Cats in Kuwait, El-Azazy, Osama M. E., Abdou Nadra-Elwgoud M. I., Khalil Amal I., Al-Batel Maha K., Henedi Adawia A. M., and Tahrani Laila M. A. , Global Veterinaria, Volume 16, Issue 1, p.111-118, (2016) cestode-trematode-stray_cats-kuwait.pdf