Nadra-Elwgoud Abdou

Feline Coronavirus (FCoV) is a worldwide viral infection of felids. The disease is usually asymptomatic, but it can cause mild diarrhoea; however, few numbers of cases may develop a severe systemic disease known as feline infectious peritonitis (FIP). This study aims to determine the prevalence of FCoV shedding in the faeces of stray cats in Kuwait and detect antibodies against FCoV in their serum. Histopathological analyses and RT‑PCR were used to prove cases of FIP. A total of 178 cats were examined for the presence of FCoV in their faeces using a rapid immunochromatography (IC) test. Anti‑FCoV Antibody (Anti‑FCoV Ab) was detected in their serum using ELISA. Eleven samples were tested using RT‑PCR to confirm positive cases. The prevalence of FCoV faecal antigen in stray cats was 32.6%. The overall detection rate of Anti‑FCoV Ab in stray cats was 44.9%. Nine cats tested positive using the RT‑PCR test. Six out of those nine were confirmed to be FIP positive through gross and histopathological examination. The characteristic uveitis and discoloration of the irises were seen. The present study is the first report confirming FCoV infection in stray cats in Kuwait. Postmortem and histopathological lesions in cases of FIP were recorded.

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.

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.

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.

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.

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.

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.

Stray cats are a common feature roaming the streets and alleys of Kuwait; they could be a source of parasites, including trematodes, that affect humans. A survey was conducted to identify feline trematodes and throw the light on their public health significance in Kuwait. Out of 240 stray cats trapped from different localities of Kuwait from June 2011 to May 2012, 59 (24.6%) were found to be infected with 14 species of trematodes. The most common were trematodes of the genus Heterophyes, particularly H. heterophyes and H. dispar that were found in respectively 15.8% and 10.8% of the cats examined. Other trematodes recorded, with lower prevalences, were Heterophyes nocens (2.9%), Haplorchis taichui (3.8%), Stictodora sawakinensis (2.1%), Stellantchasmus falcatus (1.6%), Echinochasmus japonicus (1.6%), and Mesostephanus dottrensi (1.3%). Centrocestus cuspidatus, Galactosomum fregatae, Ascocotyle sp., Mesostephanus appendiculatus, Haplorchis yokogawai, and Pygidiopsis genata showed the lowest prevalence (0.4%) and intensity. The majority of the trematodes are recorded for the first time in Kuwait and even in the Gulf region. The study reveals that stray cats are good indicators of fish-borne trematodes in the environment. As all trematodes recovered are zoonotic, their significance to public health should be considred.

In Kuwait, stray cats were surveyed for enteric protozoan infection using fecal examination and their sera were tested for Toxoplasma gondii IgG using indirect hemagglutination test (IHAT) as well as for feline immunodeficiency virus (FIV) antibodies and feline leukaemia virus (FeLV) antibodies using ELISA. Out of 240 fecal samples examined 22 (9.2%) were found to be infected with oocysts of four species of coccidian protozoa. Isopspora felis was the most predominant enteric protozoan parasite (7.1%), followed by T. gondii (2.1%), I. rivolta (1.6), Sarcocystis was only found in one case (0.4%). Juvenile cats ( 6 months old) had higher infection rate with oocyst of enteric protozoa than older cats (p-value 0.001). Sero-survey of 240 stray cats revealed that 19.6% were positive to T. gondii IgG. Toxoplasma sero-positivity was observed in higher number of adults compared to young cats suggests that with age the risk of exposure to T. gondii increases. While concurrent retroviral infections were not found to be associated with increased risk for developing T. gondii antibodies.

Bedbug, Cimex lectularius, human infestations were reported in the State of Kuwait in the last 2 years. Eleven separate infestations from different localities were received at the Veterinary Laboratories indicating that bedbug is widespread in the State of Kuwait. There was circumstantial evidence to suggest the transfer of bugs with recent immigrants or used furniture. The spread of infestation can be attributed to the increase in migrant labor and their mobility inside the country. The increase in reported cases appears also consistent with a worldwide increase in bedbug infestations.

In opposite to most countries, avian influenza virus H5N1 became endemic in Egypt. Since, its first emerge in 2006 in Egypt, the virus could infect different species of birds and animals and even human. Beside the great economic losses to the local poultry industry in Egypt, the virus infected 166 confirmed human cases, 59 cases ended fatally. In the present study, the persistence of the avian influenza in the Egyptian environment was studied. For this purpose, serum samples were collected from human, cattle, buffaloes, sheep, goat, horses, donkeys, swine, sewage rats, stray dogs and stray cats. The sera were collected from Cairo and the surrounding governorates to be examined for the presence of anti-H5N1 antibodies by Haemagglutination Inhibition Test (HI) and ELISA test. Clear differences in the seroprevalence were noticed among different species and also between the results obtained by both techniques indicating the difference in test accuracy. The present data indicate wide spread of the H5N1 virus in the Egyptian environment.

In Egypt, zoonotic fascioliasis is increasing. In this study, postmortum examination of 88 donkeys used as gargantuan meal in the Zoo at Giza revealed hepatic fascioliasis in 15 (17.05%). The serum examination for anti-Fasciola antibodies by ELISA showed positivity in 12/15 with crude worm antigen, and positivity in 14/15 with locally prepared Fasciola excretory-secretory (Fges) antigen. The zoonotic role of animal fascioliasis was discussed.