nashwa khalifa | Benha University (original) (raw)

Papers by nashwa khalifa

A B S T R A C T A total 1006 animals (300 cattle, 300 buffaloes, and 300 sheep and106 goats) were... more A B S T R A C T A total 1006 animals (300 cattle, 300 buffaloes, and 300 sheep and106 goats) were selected from private farms and suspected to suffer from brucellosis from different localities in Gharbiya governorate, as well as 50 rats (31Rattus rattus &19 Rattus norvegicus) and 15 stray dogs were collected from the same localities associated with examined animals. In addition, 160 persons suffering from fever suspected to be brucellosis were collected (80 workers contact with examined animals and 40 from fever hospitals). Serological tests were carried out by using Rose Bengal plate (RBPT), Buffered Acidified plate test (BAPAT), Complement Fixation test (CFT), Tube Agglutination test (TAT) and 2-Mercapto-Ethanol test (2-MET). The results showed that the percentage of positive reactors were 9%, 7.3%, 9.3% 8.5%, 8% and 0% using RBPT in cows, buffaloes, sheep, goats, rats and dogs respectively. Meanwhile the percentage of positive reactors using BAPAT was 9.6%, 8.3%, 10.7%, and 9.6% in cows, buffaloes, sheep and goats and by using CFT the percentage was 9.3%, 8%, 10.3% and 10.3% in previously examined animals. Also the result of TAT was 8% in rats and 0.0% in dogs. The occurrence of brucellosis in sheep and goats was higher than cows and buffaloes. Finally, the results in humans were 13.1%, 11.3% and 10% by using RBPT, TAT and 2MET respectively. The incidence of brucellosis was higher in males (14.9%) than females (3.8%) and higher in humans aged between 20-30 years.

A B S T R A C T A total of 127 specimens (13 aborted foeti, 46 milk samples, 37 lymph nodes, 14 l... more A B S T R A C T A total of 127 specimens (13 aborted foeti, 46 milk samples, 37 lymph nodes, 14 livers, 14 spleen and 6 vaginal discharges) were collected and examined for isolation and typing of Brucella microorganism. The results detected 15 strains (5 aborted foeti, 4 milk, 5 lymph nodes and 1 spleen) were detected and typed as Br. melitensis biovar 3. Application of PCR test for rapid identification of Brucella strains which isolated from lymph nodes five of naturally infected animals (two cattle, one buffaloes, one sheep and one goat) revealed that all samples were reacted positively with Br. melitensis specific DNA products with a molecular size of 731 pb. On sequencing, the Nucleotide sequence alignment of obtained sequences with other Brucella strain indicated that the obtained isolate have high identity with Br. melitensis biovar 3. The bacteriocidal activity of tested disinfectants against isolated Br. melitensis strain at variables concentration revealed that halogen showed highest bactericidal activity followed by QACs and phenolic while alkaline wasthe lowest effect.

Foot-and-mouth disease virus (FMDV) SAT2 serotype is endemic

Query (Q) fever is an ubiquitous zoonosis caused by Coxiella burnetii. The present study was carr... more Query (Q) fever is an ubiquitous zoonosis caused by Coxiella burnetii. The present study was carried out to determine the prevalence of C. burnetii in apparently healthy sheep, goats and farm workers. Raw milk and serum samples were randomly collected from 200 sheep, goats (100 of each) and 30 farm workers from Qaluobia governorate, Egypt during 2014/2015. The milk and serum samples were investigated for IgG antibodies against C. burnetii phase II antigen by indirect immunofluorescent antibody test (IFAT). The seropositive samples were confirmed by touch –down PCR with specific primers which amplify transposon-like region of C. burnetti. The results showed that antibodies against C. burnetii in sheep raw milk and sera were 17% and 23% respectively, in goat raw milk and sera were 19% and 27% respectively and in human was 23.3%. PCR targeting IS1111 gene confirmed the presence of C. burnetti DNA in sheep and goats raw milk and sera were 82.4%, 89.5%, 91.3% and 85.2% respectively and in farm workers was 57.1%. These results proved that the apparently healthy sheep and goats are an important reservoir of C.burnetii infection. The farm workers constitute an occupational risk group for C. burnetii infection, for their contact with infected livestock.

In April 2014 foot-and-mouth disease virus (FMDV) affected water buffaloes (Bubalus bubalis) aged... more In April 2014 foot-and-mouth disease virus (FMDV) affected water buffaloes
(Bubalus bubalis) aged from 3-5 years in Qalyubia, Egypt. The aim of the
present study was to diagnose FMDV molecularly and biochemically. Blood
samples were collected from buffaloes suffering from characteristic clinical
signs of FMDV infection as fever, profuse ruby threads salivation, ulcer on
muzzle, vesicles on foot and lameness. Blood samples, tongue epithelium
and vesicular fluid were evaluated by real time RT-qPCR for the diagnosis of
FMDV using different probes and primers of universal (3D) gene and VP1
gene for serotypes A, Iran O, Asia and SAT2. The positive sample confirmed
by one step reverse transcription polymerase chain reaction (RT-PCR). This
resulted in the identification of a SAT2 serotype was the causative agent and
the amplified RNA virus resulted in 716bp. Serum samples of positive PCR
infected animals compared with apparently healthy control group was used
to determine the concentration of aspartate amino transferase (AST), alanine
amino transferase (ALT), alkaline phosphatase (ALP), albumin, total protein,
calcium (Ca), iron (Fe) and inorganic phosphorus (Ph). A level of nitric oxide
(NO) and malondialdehyde (MDA) were calorimetrically measured in serum
as markers for oxidant status. There was a significant increase (P<0.05) in
AST, ALT, ALP, Ph, NO and MDA and a significant decrease (P<0.05) in
albumin, total protein, Ca and Fe in serum of clinically affected animals. It
was concluded that FMDV serotype SAT2 circulate in Eg

The prevalence of bovine cysticercosis was established using routine postmortem inspection of 345... more The prevalence of bovine cysticercosis was established using routine postmortem inspection of 3450
carcasses of buffaloes slaughtered in 2014 in Kaliouba governorate, among which 313 (9.07%) were detected
as harbouring cysticercosis lesions using meat inspection process. The cysts were examined macroscopically
for description of their morphology and constituents and classified as viable or degenerating. Viable cysts were
microscopically confirmed for demonstration of protoscolex. Out of 100 of patients offered taenicidal drugs
examined by microscopic examination through direct and sedimentation of fecal samples, 6 (6%) were positive
for Taenia saginata (T. saginata) eggs. Histological sections of 6 gravid proglottids were identified as
T. saginata. We used a biomolecular assay targeting the HDP2 gene for developing PCR assay in 20 viable
cysts and 6 gravid proglottids. An HDP2 gene-PCR amplification product of the taeniid samples of T. saginata
is approximately 599bp. Partial sequences were generated after gel purification of PCR amplified products of
HDP2 gene with sequence analysis and subsequent phylogeny to compare these sequences to those from
known strains of T. saginata circulating globally and retrieved from GenBank. Most isolates with accession No.
KT027580 are closely related to T. saginata based on the similarity of nucleotide sequences and phylogenetic
relationships. In conclusion, this work indicated high prevalence rate of bovine cysticercosis and T. saginata,
both morphological examination of the parasite and molecular analysis using bioinformatic tools identified the
metacestode and revealed typical taeniid features confirmed to Taenia saginata

In this study, groups of 150 brucella free white Swiss mice, were inoculated with different oily ... more In this study, groups of 150 brucella free white Swiss mice, were inoculated with different oily
adjuvanated brucella melitensis subunit vaccines alone or both and then were challenged with Br.
melitensis virulent strain for the evaluation of the immune response of each subunit vaccine which was
judged by testing this potency through spleen to body weight ratio and the number of brucellae per gram
spleen. Also two groups of brucella free guinea pigs were inoculated with a combination of different
oily adjuvanated Brucella melitensis subunit vaccines combined with either live attenuated Br. abortus
strain 19 vaccine, inoculated conjunctivally or RB51 vaccine injected subcutaneously. Then the animals
were challenged with virulent Br. melitensis strain. Humeral and cell mediated immune responses were
evaluated. The shedding of the vaccinal strains in the different body secretions was detected along the
whole days of the experiment. Eighteen serum samples were collected from occupational group working
in the vaccine production and application for serological examination. It was found that the protection
level in the different mice animals groups was 70%, while in the vaccinated guinea pigs vaccinated with
the combined oily adjuvanated Br. melitensis subunit vaccine combined with strain 19 vaccine, was
90%, while in those combined with RB51 vaccine, the protection level was 85%. In addition, no vaccinal
strains were detected in the different body secretions of the vaccinated animals along the whole days of
the experiments. Only two cases of the occupational group were positive

Ten female local breed cows proved to be brucella free (six of them were pregnant, parturated and... more Ten female local breed cows proved to be brucella free (six of them were pregnant, parturated and lactate
during the experiment time) were subdivided into two subgroups, the first one vaccinated with
combination of HS and OMPs subunit vaccines combined with conjunctival vaccination with Br. abortus
strain 19 vaccine and the second group was vaccinated with combination of HS and OMPs subunit
vaccines beside vaccination with Br. abortus strain RB51 (S/C). From first day post vaccination to 60
days, saliva, vaginal discharge, fecal and milk samples were collected and examined for the presence of
the vaccinal strains. Also, blood samples were collected from vaccinated animals and the serum tested
serologically using RBPT, MAT and ELISA. In addition, cell mediated immune response was evaluated
using Brucellin test. The results revealed that no vaccinal strains was detected in the different body
secretions and the humoral immune response of vaccinated cows reached its peak at the 4th week post
vaccination then decreased gradually and disappeared at the end of the 10th week. Also, cell mediated
immune response revealed that cows vaccinated with the combination of HS and OMPs subunit vaccine
combined with conjunctival vaccination with Br. abortus strain 19 vaccine showed remarkable increase
in the cell mediated immune response in comparison with in cattle vaccinated with the same combination
beside subcutaneous vaccination with Br. abortus strain RB51.

A total of 2130 samples collected from diarrhea chicken, raw milk, milk products and stool of pat... more A total of 2130 samples collected from diarrhea chicken, raw milk, milk products and stool of patient
with diarrhea from Menia, Fayoum, Cairo and Qaluobya in Egypt. Samples were subjected to standard phenotypic
identification of C.jejuni, and subsequently immunofluorescent technique (IFT) identification and genetic
amplification by PCR using specific primers of hippuricase gene. The overall prevalence of Campylobacter jejuni in
intestine and liver of chicken were 40.4 % and 37.5 % respectively, 30% tape water, 4.44% raw milk, Karish cheese
and yoghurt 6.66% and 13.33% respectively and 70 (35%) children stool. The positive results of C.jejuni were
detected by IFT expressed by green fluorescence staining. PCR amplification of hipO gene of C. jejuni isolated from
the clinically diseased chicken and the environmental samples have shown identical fingerprints with human isolates
at 344bp, indicating the zoonotic hazards of Campylobacter jejuni in Egypt

The present study was carried out to screen and analyze the characteristics of antibiotic resista... more The present study was carried out to screen and analyze the characteristics of antibiotic resistance in Campylobacter strains isolated from human and chicken in the poultry farms of different localities in Egypt. A total of 340 samples were taken from human and various poultry farms and examined for bacteriologically for isolation of Campylobacter organisms. Fifty-six (16.47%) Campylobacter-positive using conventional method including 42 (12.35%) isolates for C. jejuni and 14 (4.12%) isolates for C. coli detected. Campylobacter isolates were evaluated for their antibiotic susceptibilities.
Results of Antibiogram revealed that Campylobacter isolates were resistant to one or more of the antibiotics tested. Resistance was most frequently observed against streptomycin (96.4%) amoxicillin (94.6%), doxycycline (87.5%), Ampicillin (83.9%), nalidixic acid (85.7%), erythromycin and ciprofloxacin (82.1%). C. jejuni strains were often resistant to cephalothin (35.7%) than were C. coli strains (42.8%). C. coli were sensitive to erythromycin and Streptomycin (100%). C. jejuni was an increase sensitive to amoxicillin and streptomycin (95.2%). The trend of resistance to gentamicin (28.6%) and tetracycline (50%) was observed for C. jejuni.
The present study provides an assessment of the occurrence of multidrug resistance of Campylobacter isolates from chicken samples collected from the poultry farms in different localities in Egypt. The antimicrobial resistance rates among these pathogens are clearly important in risk assessment and management. Further research is also needed to better understand the relationship between antimicrobial use in poultry and humans and the bacterial resistance in humans.

The objective of the present study was to investigate the epidemiological and genetic relationshi... more The objective of the present study was to investigate the epidemiological and genetic relationships of classical enterotoxins of S. aureus in goat’s raw milk, meat and food handlers in Toukh city in Qaluobia governorate, Egypt. A total of 100 goat, s raw milk and meat samples (50 of each) were collected from randomly distributed herds in streets for buying milk and in public markets for peddler meat. Hand and nasal swabs were collected from milkers and butchers (30 of ech). All samples were subjected for bacteriological examination for isolation and identification of S. aureus. Isolates were underwent reversed passive latex agglutination technique for detection of enterotoxigenic S. aureus. A multiplex PCR assay could successfully amplify the diagnostic DNA bands of 270bp, 165bp, 69bp and 306bp of genes for staphylococcal enterotoxins A, B, C, and D respectively. PCR was applied on the serologically identified 16 (20.25%) isolates out of 79 S. aureus which isolated from the examined goat’s food samples and human handlers by using one universal forward and reverse primers, specific for each individual toxin gene. None of the samples was positive for SEE indicating the zoonotic and genetic relationships

Hydatidosis is one of the most important parasitic zoonoses and remains a public health and econo... more Hydatidosis is one of the most important parasitic zoonoses and remains a public health and economic
problem all over the world. The hydatid cysts were collected from slaughtered cattle and sheep in
Toukh abattoir, Kaliobia governorate, Egypt. Cyst fluid was obtained from hepatic and pulmonary
cysts for demonstration of protoscolices and hooklets. The prevalence of infection of hydatid cyst was
12.71% and 7.87% among examined cattle and sheep respectively, 42.66% and 38.46% had hydatid
cysts in liver respectively, while the infection rate was 36% and 46.15% in the lung respectively. The
rate of fertile cysts was found to be 32 (61.53%) in liver and 33(64.70%) in lung of slaughtered cattle
and sheep. PCR amplification was used for identification of internal transcribed spacer gene 1 (ITS1)
of fertile hydatid cysts obtained from cattle and sheep by using specific primer. The amplified DNA
fragment was further analyzed by PCR mediated restriction fragment length polymorphism (PCRRFLP)
using two restriction enzymes (MSP1 and RSA1). The PCR yielded similar amplified DNA
band of the same molecular size marker at 1115 bp in different isolates of Hydatid. No band variation
of ITS1 gene could be detected by PCR- RFLP by using two restriction enzymes. Amplification
product of ITSI after digestion with MSP1 showed at 661 bp and 406 bp, while those restricted with
RSA1 enzyme appeared at 745 bp and 360 b

This research have been done by direct questionnaire with house wives and student who indirect co... more This research have been done by direct questionnaire with house wives and student who indirect contact
with backyard poultry in Monofyia Governorate. Rapid avian influenza antigen detection kit test has the
characteristics of two commercially available rapid antigen tests for highly pathogenic avian influenza.
It reflected that the majority of housewives who buy poultry for house breeding from the Beachcomber
was (66.2%), followed by farm (33.5%). This percentage is due to economic factors and majority of
nannies do not have enough money to buy chickens from the farm, most of house wives breed bird away
from living area (44.75%) and (55.25%) near to living area ,and (49.8%) mix between different species.
Veterinary extension and community interventions have great effect in changing knowledge and practice
toward avian influenza

In this study, 200 fish samples of Oreochromis niloticus and Clarias gariepinus (100 of each) wer... more In this study, 200 fish samples of Oreochromis niloticus and Clarias gariepinus (100 of each) were
collected from different fish markets at Qalyoubia province. In addition, 100 skin swabs were
collected from fish sellers (60) and house wives (40) from the same localities. The objective of this
study is to detect the occurrence of some bacterial zoonotic microorganisms from market fish such as
Staph. aureus , Salmonella spp., E. coli , and Streptococci and to detect the effect of heat treatment as
frying and grilling on survival of inoculated (10)4Staph.aureus ,(10)5Salmonella typhimurium and
(10)6 E.coliO157H7 in both fish spp. In this study, it was evident that C. gariepinus samples had a
significantly higher bacterial isolates (9.8%) than O. niloticus samples (6%). Among the isolated
bacteria Streptococci was detectable at higher percentage (13.5%) followed by Salmonella spp
(11.5%), then Staph. aureus (4.5%) and the lowest isolates were E. coli (2%).The higher percentage of
bacterial isolates was recovered from surface samples than those isolated from muscle samples of
both of the examined fish spp. Hand swabs of fish handlers revealed that Staph. aureus ,Salmonellae ,
E. coli and Streptococci were isolated at percentages of 35% , 35% , 20 % and 35% respectively from
fish sellers compared to 37.5% , 25% , 37.5% and 50% respectively from house wives .Also these
results showed that frying of fish lead to total destroying of inoculated pathogens in both fish spp. at
different weights . While grilling not efficient as frying as it kill all the inoculated pathogens in O.
niloticus but not in large sized C. gariepinus. The public health importance of isolated microorganisms
and suggested hygienic measures were discussed.

is still one of the main causes of bacterial gastroenteritis worldwide. This work was done to inv... more is still one of the main causes of bacterial gastroenteritis worldwide. This work
was done to investigate the fingerprinting of isolated from chicken, dairy cattle and
human. Fecal samples were collected from 100 diarrheic chickens and dairy cattle (50 of each) as well as 50 stool
samples from patients with diarrhea were subjected to standard isolation and identification of
. DNA of isolates was amplified using specific primers of hippuricase gene. The prevalence of
was 18(36%) in chicken, 16 (32%) in dairy cattle and 11(22%) in patients with diarrhea.
PCR analysis produced identical bands at 344 bp in all isolates, indicating the role of chicken and dairy cattle
in human infection

Salmonellosis is one of the most important zoonotic bacterial pathogen of food-borne infection al... more Salmonellosis is one of the most important zoonotic bacterial pathogen of food-borne
infection all around the world.The present study was carried out to report the prevalence of the serotypes and
genetic types of salmonella among broiler chickens,raw chickens meat and patients suffer from food poisoning
signs in Toukh Egypt.Samples collected from (50) diarrheic broiler chicken,(50) raw frozen chickens meat and
(30) diarrheic patients with food poisoning signs were bacteriologically and serologically processed for
identification of Salmonella. Isolates were subjected to multiplex-PCR using specific Salmonella
primers.The prevalence of Salmonella spp was 7(14%), 2(4%) and 3(10%) in chickens,raw chickens meat and
patients respectively. TheSalmonella Isolates were serologically identified as 7(58.33%) and 5[41.66%]
S.enteritidis and S.typhimurium respectively.The antigenic formula of serovar S.enteritidis has somatic antigen
O: 1,9,12 and phase I (g,m) and phase II [1,7] flagellar antigen (H),While serovars S.typhimurium have the O:
1,4, (5),12and phase I and phase II.Multiplex-PCR yield similar diagnostic amplified DNA bands of molecular
size marker at 250bp in tested S.enteritidis and 620bp in examined S.typhimurium,indicating the zoonotic
potential of the organism and the role of chicken and chicken meat as sources in the epidemiology of the human
salmonellosis.

Cryptosporidium parvum is a common intestinal parasite which is associated with severe acute diar... more Cryptosporidium parvum is a common intestinal parasite which is associated with severe acute diarrhoea
in humans and animals. This work aimed to determine Cryptosporidium sp. in positive faecal
samples by using polymerase chain reaction (PCR). Cryptosporidium oocysts were isolated from 10
out of 20 Egyptian stray dogs with diarrhoea. The zoonotic C. parvum was found to be present in two
isolates which showed successful amplification of a specific DNA fragment at 550 bp with C. parvum
outer wall protein (COWP) gene amplicon using two specific primers CRY-15 and CRY-9. This is
the first investigation on the presence of C. parvum among Egyptian stray dogs and it has pointed to
the existence of a genotype that may play an important role as a source of human and farm animal
cryptosporidiosis

The aims of the present study were to diagnose toxoplasmosis in pregnant ewes and women serologic... more The aims of the present study were to diagnose toxoplasmosis in pregnant ewes and women serologically and molecularly, treat naturally infected ewes, and diagnose congenital toxoplasmosis. Blood samples were taken from 30 and 60 pregnant ewes and women, respectively, and used for diagnosis of toxoplasmosis through Latex agglutination test (LAT). Seropositive samples were confirmed by PCR for detection of acute infection. Ten infected pregnant ewes were classified into two groups. The first group was treated with sulfadimidine 33.3%, 200 mg (0.6 ml) / kg.b.wt, and the other group was treated with normal saline. At titers ≥ 1:64, serological diagnosis indicated that 16 (53.33%) ewes and 29 (48.3%) women were seropositive and the seroprevalence increased in older ewe and younger women. Positive LAT samples were used for amplification of DNA and showed bands at 193 bp, analogues to that of the RH strain in 12 (40%) and 15 (25%) blood samples of ewes and women, respectively. All treated ewes with sulfadimidine 33.3% delivered healthy lambs with normal gestation period, whereas untreated ewes delivered 4 abortuses and 3 stillbirths. The tissue cysts were demonstrated microscopically in stained smears from tissues of dead fetuses. Local strain of T. gondii was isolated through intra peritoneal injection in mice from tissues of abortuses and stillbirths and maintained in the lab. The DNA of both RH (a virulent

The pulmonary and hepatic hydatid cyst fluids were collected from 540 slaughtered camels and 5 hu... more The pulmonary and hepatic hydatid cyst fluids were collected from 540 slaughtered camels and 5 human cases in Qalyubia Governorate, Egypt. The prevalence of infection of cystic echinococcosis among camels was 120 (22.2%). The fertility rates of the isolated cysts form camels and humans were 64.5 and 100%, respectively. A nested polymerase chain reaction was used for amplification of mitochondrial NADH 1 gene of Echinococcus granulosus complex in fertile cysts obtained from camels and humans, respectively. Two pairs of primers (EGL1 and EGR2) and (EGL3 and EGR4) were used in 2 amplification steps. First, the outer pair of primer originated from a highly conserve region of NADH1 gene generate a primary 435 bp PCR product. Second, a pair of internal (nested) primer (EGL3 and EGR4), designed to the annealing site of primers (EGL1 and EGR2) yield similar diagnostic amplified DNA bands of molecular size marker at 276 bp in all examined cysts obtained from camels and humans indicating a zoonotic relationship. This study confirms similar fingerprinting patterns of Echinococcus granulosus complex in camels and humans in Qalyubia Governorate, Egypt. Nested PCR for diagnosis of E. granulosis had been used for the first time in Egypt, as far as we know.

Campylobacteriosis -caused principally by Campylobacter jejuni (C. jejuni) and Campylobacter coli... more Campylobacteriosis -caused principally by Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) -is among the main causes of bacterial gastroenteritis worldwide. This work was done to investigate the molecular characterization of zoonotic C. jejuni and C. coli isolated from fecal samples of beef cattle, retail beef meat and beef liver and stool of children with diarrhea. Fecal samples were collected from 50 apparently healthy cattle, 60 of retail beef meat and beef liver (30 of each) as well as 50 stool samples from pediatric diarrhea were subjected to standard isolation and phenotypic identification of Campylobacter isolates. The prevalence of Campylobacter isolate was 17(34%) in fecal sample of cattle, 5(16.66%) beef meat, 8(26.66 %) beef liver and 13 (26%) in pediatric diarrhea. Out of 43 identified isolates, 26(60.46%) C. jejuni isolates were higher than 14(32.55%) C. coli, two samples were mixed infection and one Campylobacter upsaliensis. A multiplex-PCR method was developed for the detection of C. jejuni and C. coli. Primers were the hippuricase gene (hipO) characteristic of C. jejuni, a sequence partly covering an aspartokinase gene (asp) characteristic of C. coli and a universal 16S rDNA gene sequence serving as an internal positive control. All Campylobacter isolates expressed identity with 16S rDNA (genus specific gene) at 1062 pb. Multiplex PCR demonstrated one false-positive and one false-negative hippurate activity test. PCR method was incapable to identify biochemically identified C. upsaliensis. Amplification of hipO gene of C. jejuni and aspgene of C. coli isolated from cattle, beef and liver have shown identical fingerprints with human C. jejuni and C. coli at 344bp and 500bp respectively, indicating the public health importance of the isolates.

Foot-and-mouth disease virus (FMDV) SAT2 serotype is endemic