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- Zeitschrift
- eISSN
- 1336-9083
- Erstveröffentlichung
- 22 Apr 2006
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- 4 Hefte pro Jahr
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- Englisch
In Egypt, camels (
Cystic echinococcosis (CE) or hydatidosis is one of the most dangerous zoonotic diseases, it can be fatal if untreated. It occurs due to infection with the larval stage of the common dwarf dog tapeworm,
Conventionally, different isolates of
Detailed evidence about the diversity of
Until now, however, few studies have reported the presence of genotypes of the
A cross sectional study was conducted in Aswan city, which is located at a latitude of 24° 5′ 20.18″ N and a longitude of 32° 53′ 59.39″ E. Aswan is famously for its beautiful Nile Valley scenery, important archaeological sites and peaceful atmosphere. The climate is warm throughout the year, making it an ideal winter destination. Furthermore, agriculture is the main source of employment opportunities in the province, with approximately 29 % of the province’s population working in agriculture.
The sample size was calculated according to the method previously described by Charan (2013). The parameters of the formula (n=Z2×P (1-p) /d2) were as follows: Z = Is standard normal variate at 5 % type 1 error (P<0.05), it is 1.96, P= the expected prevalence of HC based on previous studies was 9.7 % (Zienab 2021), and d = is ±5 % desired level of precision. The formula was n = (1.96)2 × 0.097 × (1 – 0.097)/0.05. The required sample size n was determined to be 110 camels. To determine the prevalence of hydatid cyst infection in the camels of the southern region of Egypt, the visceral organs (liver, lungs, kidneys, and spleen) of 110 slaughtered camels were inspected over the course of one year from June, 2018 to May, 2019 in Daraw Abattoir, Aswan, Egypt. During the sampling gender, age, and the seasonal dynamics were recorded.
All organs or tissues containing HCs were collected and all HCs found were carefully removed and separately collected, kept in sterile saline solution, 70 % ethanol and transported within 2 hours to the Parasitology laboratory, Faculty of Veterinary Medicine, South Valley University in ice box for further cyst characterization to assess their status and for genetic strain typing of
Five samples of hydatid fluids were collected from the lung cysts of the screened hosts (camels and cattle), approximately 25ml of the collected fluids (HCF) was centrifuged at 15000 rpm at 4 ºC for 30 min and the supernatant was analysed for various biochemical parameters. The flame photometry method was utilized to measure the amount of Sodium and Potassium, copper and magnesium by the spectrophotometry method. Creatinine, Calcium and Protein were analyzed by the colorimetric method. The measurements of Glucose, cholesterol, triglycerides, and urea was made by enzymatic methods (SYNCHRON CX 7 PRO, USA 2009). These parameters were estimated by a commercially available diagnostic Kits from Sigma, Germany. Each electrolyte and biochemical profile was prepared and quantified according to the manufacturer’s instructions.
Statistical significance differences were assessed with a Chisquare using a statistical package program (Sigma Plot version 11.0). Statistical significance was considered with P values < 0.05.
DNA was successfully extracted from two fertile HCs out of fourteen cysts using QIAamp DNA Mini Kit (Qiagen, Germany, Catalogue no.51304,) according to the manufacturer’s recommendation. DNA was stored at -20 °C until being used for DNA amplification. For all DNA extracts, PCR were conducted on the gene encoding
GenBank accession numbers of cox1 and nad1 of
| Accession No. for NAD1 | Locality | Host origin | Genotype | Accession No. for Cox1 | Locality | Host origin | Genotype |
|---|---|---|---|---|---|---|---|
| MW183240 | Egypt | Camel | G7* | MW173485 | Egypt | Camel | G7* |
| MW183239 | Egypt | Camel | G7* | MW173484 | Egypt | Camel | G7* |
| MH301013 | France | Pig | G7 | MH301013 | France | Pig | G7 |
| KX010897 | Hungary | Pig | G6/7 | MH301019 | Italy | Pig | G7 |
| KX010890 | France | Pig | G6/7 | MH301010 | France | Pig | G7 |
| KX010889 | Namibia | Oryx | G6/7 | MH301009 | France | Pig | G7 |
| KX010886 | Sudan | Camel | G6/7 | MH301008 | France | Pig | G7 |
| MH301016 | France | Pig | G7 | MH300939 | Sudan | Camel | G6 |
| KX510135 | Serbia | Pig | G6/7 | KX010833 | Kenya | Camel | G6/7 |
| KX231668 | Armenia | Pig | G6/7 | MT166286 | Nigeria | Camel | G6 |
| KX010880 | Ethiobia | Cattle | G6/7 | MT166284 | Nigeria | Cattle | G6 |
| MT525964 | Kenya | Cattle | G6/7 | MH301021 | Ukraine | Pig | G6 |
| MT166290 | Nigeria | Camel | G6 | MH301006 | Poland | Pig | G7 |
| MT166289 | Nigeria | Cattle | G6 | MH301005 | Poland | Homosapiens | G7 |
| JN191326 | Egypt | Donkey | G4 | JN191319 | Egypt | Donkey | G4 |
| MK644934 | South Korea | Human | MK644934 | South Korea | Human |
*Sequences generated in the current study
All applicable institutional, national and international guidelines for the care and use of animals were followed regarding the National Research Ethics Committee of the South Valley University and Veterinary authorities in South Valley University, Egypt.
Out of 110 dromedaries examined, only 11 were found to be infected with one or more cysts giving an overall prevalence of 10 %. In respect to cyst distribution, the current finding declared that lungs were the most commonly infected organs. The current survey revealed that infection varied according to the camel’s age, with the highest prevalence of infection in older animals of age 3 years and over (12.3 %, 8/65), while animals with mostly 1 – 2 years have less infection rate (6.7 %, 3/45). The statistical analysis, however, confirmed that these differences were not statistically significant (
Fig. 1
a, b) Calcified cyst; c) Protoscoleces; d) Multiple fertile cysts in the lung (yellow arrow head).
Additionally, the fully calcified cysts were observed in the liver with estimated fertility percentage of 8.6 % (3/35) and the rest cysts was found sterile (5.7 %, 2/35) as depicted in Fig. 1. Concerning the cyst viability, the current study displayed that the viability rate of protoscoleces was 60.7 % from fertile lung cysts.
Additionally, the current finding showed that the size of hydatid cysts collected from lungs and liver ranged from 1 – 9 and 1 – 2 cm in diameter, respectively. Furthermore, the average weight of the recovered cysts from lungs and livers were 70.9 g (7 – 135 g) and 25g (15 – 35g), respectively.
Seasonal dynamics of hydatid cysts was also taken under consideration. Data analysis revealed that camels had a distinct pattern of hydatidosis in different seasons: winter (45.5 %)> summer (27.3 %)>autumn (18.2 %)>spring (9.1 %) signifying the infection throughout year. The statistical analysis, however, confirmed that these differences were not statistically significant (χ2
The comparison between the biochemical profile of HCF isolated from intermediate hosts including camel and cattle was depicted in Table 2. From this table, it was observed that, the mean value of total protein, glucose, urea, cholesterol, magnesium, potassium, copper and creatinine was higher in HCF collected from camels as compared to the cystic fluid collected from cattle, while the mean value of calcium, triglyceride, sodium was found lower in the camel HCF as compared to cattle HCF.
Comparative biochemical analysis of hydatid fluids collected from dromedary camels and cattle infected with cystic echinococcosis (mean ± SE, n = 5).
| Biochemical profiles | Host | |
|---|---|---|
| Total protein (g/l) | 3.76 ± 0.73 | 2.42 ± 0.01 |
| Glucose (mmol/L) | 3.36 ± 0.16 | 2.58 ± 0.47 |
| Urea (mmol/L) | 5.21 ± 0.17 | 4.36 ± 0.5 |
| Cholesterol (mmol/L) | 2.94 ± 0.61 | 2.19 ± 0.31 |
| Creatinine (μmol/L) Calcium (mmol/L) | 101.22 ± 3.09 2.937 ± 0.03 | 80.67 ± 0.66 3.224 ± 0.02 |
| Triglycerides (mmol/L) | 0.039 ± 0.01 | 0.186 ± 0.003 |
| Sodium (mmol/L) | 111.35 ± 5.45 | 118.09 ± 0.54 |
| Potassium (mmol/L) | 13.92 ± 7.41 | 10.05 ± 0.05 |
| Magnesium (mmol/L) | 1.113 ± 0.33 | 1.019 ± 0.02 |
| Copper (ppm) | 0.42 ± 0.11 | 0.338 ± 0.016 |
As far as our knowledge, the present study genetically showed the presence of
PCR amplification with specific primers generated two different bands of 450-bp and 500-bp in regard to PCR amplification of
Fig. 2
Agarose (1.5%) gel electrophoresis of PCR-products of
L: ladder, Pos.: positive control, Neg.: negative control, 1, 2, 3, 4and 5 samples (all +ve).
Fig. 3
Agarose (1.5%) gel electrophoresis of PCR-products of
L: ladder, Pos.: positive control, Neg.: negative control, 1, 2, 3,4and 5 samples (all +ve).
The Blast analysis results of the sequenced data indicated the existence of one isolate belonging to G7 genotype (pig strain,
Sequencing of our samples revealed mutations in three nucleotides generating a change at the level of 180, 381 and 387 nucleotide, where a G replaced by A for G7 isolate from France. In addition, our isolate revealed three other mutations at the levels of 180, 381 and 387 nucleotides, where G substituted A and further one mutation was occurred at level 336, where C replaced by T. while, the fifth mutation at position 432, where A substituted with G for G6 isolate from Nigeria as shown in Table 3.
Fig. 4
Genetic relationships of the obtained genotypes from camel in the present study and reference sequences related genotypes of
Difference in bases at the
| Accession No. | Genotype | Nucleotide Substitution No. | Substitution/Position | Substitution/Position | Substitution/Position |
|---|---|---|---|---|---|
| (MH301013) | G7 (Pig, France) | Three | G to A 180/381/387 | - | - |
| (KX010880) | G6/7 (Cattle, Ethiobia) | Four | G to A 180 / 381 | C to T 336 | A to G 432 |
| (MT166290) | G6 (Camel, Nigeria) | Five | G to A 180 /381/387 | C to T 336 | A to G 432 |
Phylogenetic analysis revealed a robust tree associating our isolate of G7 genotype with a variety of G7 genotype (common pig strain) sequences from different geographical regions of the world, although it was more genetically related to the France isolate.
The current survey declared that the overall prevalence of HC was 10 % among slaughtered camels. The current findings were nearly close to the previous reports carried out in Assiut, Egypt (9 %) (Khalifa et al., 2014). Other reports in several Egyptian governorates reported relatively higher prevalence rates among slaughtered camels e.g. 16.25 % in Qalyubia (Mahmoud, 2012) and 39.5 % Beni-Suef (El-Dakhly et al., 2019). However, the infection rates of 2.5 %, 7.7 %, 5.6 % and 5 % and 8.32 % has been reported for camel hydatidosis from Mansura city (Haridy et al., 2006), Assiut and Aswan (Dyab et al., 2005), Giza (El-Dakhly et al., 2019) and Aswan (Omar et al., 2013 & Dyab et al., 2018), respectively. The infection rate of 59 %, 29.7 % 32.8 %, 30.1 % and 61.4 % has been reported from Sudan (Omer et al., 2010; Ibrahim et al., 2011), Saudi Arabia (Ibrahim, 2010), Mauritania, and Kenya (Njoroge et al., 2002), respectively. Additionally the present finding was also greater than worldwide reports done in Libya (3.6 %) and Tunisia (6.5 %), respectively (Kassem & Gdoura, 2006; Lahmar et al., 2004). This variation in prevalence could be due to several factors including differences in environmental conditions that are conducive for the perpetuation of the parasite, social activities and culture, approach towards dogs and difference in husbandry and hygienic system. Other reasons like the personal behavior of workers about the elimination of infected offal in abattoirs and the nature of the pasture may contribute to this variation (Salih et al., 2011; Abdelbaset et al., 2021).
The present study revealed that the prevalence of HCs varies greatly among screened age groups; those with age of 3 years and over (>3) were highly infected (12.3 %) with
Fig. 5
Genetic relationships of the obtained genotypes from camel in the present study and reference sequences related genotypes of
The current figures indicated that the rate of infection in lungs (72.7 %) was higher than that of the liver (27.3 %). Our observation was in accordance with those noticed in camels in several African countries, such as Mauritania (Bardonnet et al., 2003), Sudan (Ibrahim et al., 2011), and Ethiopia (Salih et al., 2011). This might be due to the fact that camels are slaughtered at an older age, during which period the liver capillaries are dilated and most oncospheres pass directly to the lungs; additionally, it is possible for the
The fertility of cysts is an important factor affecting the spread of
The current data revealed a vital seasonal fluctuations for hydatid cyst in camel among the various seasons. The peak infection rate was observed in winter and summer seasons. Likewise, several previous reports found a seasonal variation in infection prevalence, with the highest trend of occurrence in winter (Daryani et al., 2007; Almalki et al., 2017). The seasonal variation in infection prevalence maybe related to the differences in the environmental conditions that are responsible for the dissemination of the parasite, the availability of infected final hosts, and the nature of the pasture among seasons (Ernest et al., 2009).
In this study, the existence of
The chemical composition of hydatid cyst fluid (HCF) plays a significant role in the immunology, metabolism, physiology and existence of the cyst (Muhsin et al., 2015). Mover, the biochemical analysis of HCF collected from different intermediate hosts can provide evidence as an additional diagnostic tool to distinguish different circulating isolates in the area, and can help to identify common isolates that circulate in different hosts (Muhsin et al., 2015). The current study declared a quantitative variation in the biochemical profiles of hydatid fluids of camels from cattle, proposing the existence of camels strains of
The current sequence analysis of two camel hydatid cysts revealed the presence of one genotype of
Globally, several studies have been conducted on camel isolates from different countries, mainly in Africa. The previous report carried out in Mauritania found that G6 genotype in all twenty camel isolates (Bardonnet et al., 2002). In another DNA sequence-based revision in North Africa, all camel isolates belonged to G6 genotype (Bart et al., 2004). Earlier investigation, revealed that all camel isolates belonged to G1 and G6 genotypes in Libya and Algeria, respectively (Tashani et al., 2002; Bardonnet et al., 2003). Furthermore,
In Egypt, previous studies upon nuclear (Khalifa et al., 2014) and mitochondrial genes (Abdel-Aaty et al., 2012) indicated the presence of G6 genotype as the only predominant genotype in slaughtered camels in Qalyubia and Sohag Governorate, respectively. Another investigation demonstrated the presence of
The association with distinct host species, largely separate geographical distribution and limited rate of cross-fertilization are the main factors that have limited the gene flow between genotypic groups. G6 is commonly involved in a cycle between goats/camels–dogs and G7 mainly pigs–dogs, these two also share some overlap in their life cycles as both can infect the same intermediate hosts. Furthermore, G6/G7 share the same final host (dog) cross-fertilization has apparently been frequent enough to guarantee that G6/G7 have not diverged (Laurimäe et al., 2018).
In conclusion, the current findings report for the first time the dominance of
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Difference in bases at the nad1 locus of E. granulosus s.l.
| Accession No. | Genotype | Nucleotide Substitution No. | Substitution/Position | Substitution/Position | Substitution/Position |
|---|---|---|---|---|---|
| (MH301013) | G7 (Pig, France) | Three | G to A 180/381/387 | - | - |
| (KX010880) | G6/7 (Cattle, Ethiobia) | Four | G to A 180 / 381 | C to T 336 | A to G 432 |
| (MT166290) | G6 (Camel, Nigeria) | Five | G to A 180 /381/387 | C to T 336 | A to G 432 |
Comparative biochemical analysis of hydatid fluids collected from dromedary camels and cattle infected with cystic echinococcosis (mean ± SE, n = 5).
| Biochemical profiles | Host |
|
|---|---|---|
| Total protein (g/l) | 3.76 ± 0.73 | 2.42 ± 0.01 |
| Glucose (mmol/L) | 3.36 ± 0.16 | 2.58 ± 0.47 |
| Urea (mmol/L) | 5.21 ± 0.17 | 4.36 ± 0.5 |
| Cholesterol (mmol/L) | 2.94 ± 0.61 | 2.19 ± 0.31 |
| Creatinine (μmol/L) Calcium (mmol/L) | 101.22 ± 3.09 2.937 ± 0.03 | 80.67 ± 0.66 3.224 ± 0.02 |
| Triglycerides (mmol/L) | 0.039 ± 0.01 | 0.186 ± 0.003 |
| Sodium (mmol/L) | 111.35 ± 5.45 | 118.09 ± 0.54 |
| Potassium (mmol/L) | 13.92 ± 7.41 | 10.05 ± 0.05 |
| Magnesium (mmol/L) | 1.113 ± 0.33 | 1.019 ± 0.02 |
| Copper (ppm) | 0.42 ± 0.11 | 0.338 ± 0.016 |
GenBank accession numbers of cox1 and nad1 of Echinococcus genotypes used in phylogenetic analysis in the current study
| Accession No. for NAD1 | Locality | Host origin | Genotype | Accession No. for Cox1 | Locality | Host origin | Genotype |
|---|---|---|---|---|---|---|---|
| MW183240 | Egypt | Camel | G7* | MW173485 | Egypt | Camel | G7* |
| MW183239 | Egypt | Camel | G7* | MW173484 | Egypt | Camel | G7* |
| MH301013 | France | Pig | G7 | MH301013 | France | Pig | G7 |
| KX010897 | Hungary | Pig | G6/7 | MH301019 | Italy | Pig | G7 |
| KX010890 | France | Pig | G6/7 | MH301010 | France | Pig | G7 |
| KX010889 | Namibia | Oryx | G6/7 | MH301009 | France | Pig | G7 |
| KX010886 | Sudan | Camel | G6/7 | MH301008 | France | Pig | G7 |
| MH301016 | France | Pig | G7 | MH300939 | Sudan | Camel | G6 |
| KX510135 | Serbia | Pig | G6/7 | KX010833 | Kenya | Camel | G6/7 |
| KX231668 | Armenia | Pig | G6/7 | MT166286 | Nigeria | Camel | G6 |
| KX010880 | Ethiobia | Cattle | G6/7 | MT166284 | Nigeria | Cattle | G6 |
| MT525964 | Kenya | Cattle | G6/7 | MH301021 | Ukraine | Pig | G6 |
| MT166290 | Nigeria | Camel | G6 | MH301006 | Poland | Pig | G7 |
| MT166289 | Nigeria | Cattle | G6 | MH301005 | Poland | Homosapiens | G7 |
| JN191326 | Egypt | Donkey | G4 | JN191319 | Egypt | Donkey | G4 |
| MK644934 | South Korea | Human | MK644934 | South Korea | Human |
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