Phylogenetic evidence of a possible Trichuris globulosa species complex in Arabian camels from Kuwait.

During a 1-year study, Trichuris adults were obtained after necropsy of Arabian camels (Camelus dromedarius) from a slaughterhouse in Kuwait. Morphological and molecular identification was performed to confirm the identity of the Trichuris specimens obtained from C. dromedarius. Fifteen male Trichuris specimens were selected, and molecular identification was performed using mitochondrial cytochrome c oxidase subunit I, 12S ribosomal RNA, 16S ribosomal RNA genes and the nuclear internal transcribed spacer 2 (ITS2) region. Through phylogenetic analysis, 2 distinct groups were obtained using the mitochondrial genes, where group 1 showed a close relationship to Trichuris globulosa while group 2 showed a close relationship to Trichuris ovis, providing molecular evidence of a possible T. globulosa species complex. Additionally, the nuclear ITS2 region did not provide enough resolution to distinguish between the 2 groups of Trichuris specimens. Observation of morphological characters revealed variations in the shape of the male spicule sheath, where specimens present either a globular posteriorly truncated swelling or the absence of posteriorly truncated swelling. Moreover, the variations in male spicule sheath does not corroborate with the results of molecular data, suggesting the limited use of this character for identification of T. globulosa. In conclusion, molecular analysis suggests a possible species complex in T. globulosa, with the mitochondrial genetic markers successfully differentiating between the 2 groups. The limited use of the male spicule sheath as a diagnostic character for identification of T. globulosa is suggested.

Fate of Cryptosporidium and Giardia through conventional and compact drinking water treatment plants.

Over the past three decades, a notable rise in the occurrence of enteric protozoan pathogens, especially Giardia and Cryptosporidium spp., in drinking water sources has been observed. This rise could be attributed not only to an actual increase in water contamination but also to improvements in detection methods. These waterborne pathogens have played a pivotal role in disease outbreaks and the overall escalation of disease rates in both developed and developing nations worldwide. Consequently, the control of waterborne diseases has become a vital component of public health policies and a primary objective of drinking water treatment plants (DWTPs). Limited studies applied real-time PCR (qPCR) and/or immunofluorescence assay (IFA) for monitoring Giardia and Cryptosporidium spp., particularly in developing countries like Egypt. Water samples from two conventional drinking water treatment plants and two compact units (CUs) were analyzed using both IFA and qPCR methods to detect Giardia and Cryptosporidium. Using qPCR and IFA, the conventional DWTPs showed complete removal of Giardia and Cryptosporidium, whereas Mansheyat Alqanater and Niklah CUs achieved only partial removal. Specifically, Cryptosporidium gene copies removal rates were 33.33% and 60% for Mansheyat Alqanater and Niklah CUs, respectively. Niklah CU also removed 50% of Giardia gene copies, but no Giardia gene copies were removed by Mansheyat Alqanater CU. Using IFA, both Mansheyat Alqanater and Niklah CUs showed a similar removal rate of 50% for Giardia cysts. Additionally, Niklah CU achieved a 50% removal of Cryptosporidium oocysts, whereas Mansheyat Alqanater CU did not show any removal of Cryptosporidium oocysts. Conventional DWTPs were more effective than CUs in removing enteric protozoa. The contamination of drinking water by enteric pathogenic protozoa remains a significant issue globally, leading to increased disease rates. Infectious disease surveillance in drinking water is an important epidemiological tool to monitor the health of a population.

Antischistosomal effects of green and chemically synthesized silver nanoparticles: In vitro and in vivo murine model.

Schistosomiasis is one of the most important neglected tropical diseases in Africa, caused by blood fluke, Schistosoma sp. The use of nanotechnology in the treatment of this type of disease is urgently important to avoid the unwanted side effects of chemotherapy. The present study aimed to evaluate the efficacy of green silver nanoparticles (G-AgNPs), fabricated by (Calotropis procera), comparing with both chemically prepared silver ones (C-AgNPs) and Praziquantel (PZQ) treatments. The study included in vitro and in vivo evaluations. In in vitro study, 4 groups of schistosome worms were exposed to treatments as follows: the first one with a dose of PZQ (0.2 µg/ml), the 2nd and 3rd groups with different concentrations of G-AgNPs and C-AgNPs, respectively and the last one act as a negative control group. In in vivo study, six groups of mice were infected and then treated as follows: the first one with a dose of PZQ, the second with G-AgNPs, the third with C-AgNPs, the fourth with G-AgNPs plus a half dose of PZQ, the fifth with C-AgNPs accompanied by a half dose of PZQ, and the last group acted as a positive control group. The parasitological (worm burden, egg count & oogram) and histopathological parameters (hepatic granuloma profile) were used to evaluate antischistosomal activities in experimental groups. Additionally, the subsequent ultrastructural alterations were observed in adult worms using scanning electron microscopy (SEM). Transmission electron microscopy analysis showed that G-AgNPs and C-AgNPs have 8-25 and 8-11 nm in diameter, respectively, besides, fourier transform infrared analysis (FTIR) revealed the presence of organic compounds (aromatic ring groups) which act as capping agents around the surfaces of biogenic silver nanoparticles. In in vitro experiment, adult worms incubated either with G-AgNPs or C-AgNPs at concentrations higher than 100 µg/ml or 80 µg/ml, respectively, showed full mortality of parasites after 24 h. In the infected treated groups (with G-AgNPs plus PZQ & C-AgNPs plus PZQ) showed the most significant reduction in the total worm burdens (92.17% & 90.52%, respectively). Combined treatment with C-AgNPs and PZQ showed the highest value of dead eggs (93,6%), followed by G-AgNPs plus PZQ-treated one (91%). This study showed that mice treated with G-AgNPs plus PZQ significantly has the highest percentage of reduction in granuloma size and count (64.59%, 70.14%, respectively). Both G-AgNPs plus PZQ-treated & C-AgNPs plus PZQ treated groups showed the highest similar values of reduction percentage of total ova count in tissues (98.90% & 98.62%, respectively). Concerning SEM, G-AgNPs-treated worms showed more variability in ultrastructural alterations than G-AgNPs plus PZQ-treated one, besides, worms treated with C-AgNPs plus PZQ exhibited the maximum level of contractions or (shrinkage) as a major impact.

Investigation of honey bee venom effect on the immunogenicity of foot-and-mouth disease vaccine in sheep.

Background: Foot-and-mouth disease (FMD) is one of the most highly contagious and economically significant diseases of cloven-hoofed animals worldwide. FMD virus (FMDV) is the cause of the disease. The virus has seven serological types, identified as; O, A, C, SAT1, SAT2, SAT3, and Asia1. The aim of this study enhancement of FMD vaccine immunogenicity is the unique way to control FMD in Egypt. Aim: Our research studied the effect of bee venom (BV) as simultaneously inoculated with the commercial vaccine on the immune response of experimentally vaccinated sheep in comparison with the inoculation of the vaccine alone through evaluation of the cellular and humoral immune response. Methods: Estimation of cellular immunity using phagocytic activity, phagocytic percentage, lymphocyte blastogenesis, interleukin-6 (IL-6), and interleukin-12 (IL-12) and estimation of humoral immunity using serum neutralization test (SNT) and enzyme-linked immunosorbent assay (ELISA). Result: Evaluation of the cellular immunity expressed in lymphocyte blastogenesis, phagocytic activity, phagocytic percentage, IL-6, and IL-12 showed higher levels in sheep vaccinated by the trivalent FMD vaccine (serotypes O pan Asia, A Iran O5, and SAT2/EGY/2012) with BV comparable to those induced by the vaccine alone. Following up the humoral immune response of vaccinated sheep revealed that FMDV antibodies serotypes O pan Asia, A Iran O5, and SAT2/EGY/2012 as measured by SNT and ELISA assay induced by FMD with BV were higher than those induced by inactivated FMD alone. Conclusion: The inoculation of BV with FMD vaccine simultaneously is of high benefit inducing high level of specific immunity which could be of long duration.

Intestinal parasitic infections among expatriate workers in Al-Madina Al-Munawarah, Kingdom of Saudi Arabia.

Al Madinah Al Munawarah, Kingdom of Saudi Arabia (KSA) has one of the largest number of expatriate workers. Most of them are from Sri-Lanka, Indonesia, Philippines, India and Bangladesh. These countries are considered as endemic areas for intestinal parasites. A total of 2732 stool samples were screened for intestinal parasites. Positive cases were recorded among 407 stool samples (14.9%). The common parasitic infections were encountered among 20-29 age groups (18.5%) while, the lowest infection rate was among individual ≥ 50 years (11.8%). According to the nationality, the highest infections were recorded among Pakistanis (23.2%), followed by Philippines (22.2%), Sudanese workers (18.7%), and the lowest infection rates were recorded among Bangladeshi individuals. The infected persons were farmers, food handlers and shepherds. The detected intestinal parasites were Giardia lamblia (21.9%), Entamoeba histolytica/Entamoeba coli (17.8%), Trichuris trichiura (16.2%), Ascaris lumbricoides (15.8%), hookworm (13%), Hymenolepis nana (8.9%), Strongyloides sterocoralis (3.5%), Schistosoma mansoni (2.2%), and Enterobius vermicularis (0.43%). In conclusion, the high prevalence of parasites among expatriates may produce health problem among the Saudi society due to the nature of the expatriates' work.

Effects of garlic on Schistosoma mansoni harbored in albino mice: molecular characterization of the host and parasite.

Garlic has been used for its health benefits for thousands of years. Modern research confirmed many of the healing properties of garlic, including its antiparasitic activity. This study was designed to evaluate the antischistosomal action of garlic through detecting the changes in DNA profile of Schistosoma mansoni worms and the infected mouse. Forty mice were subcutaneously infected with ~200 Schistosoma mansoni cercariae/mouse. Infected mice were divided into four equal groups: non-treated, prophylactic, therapeutic, and continuously-treated. Non-infected control and garlic-treated groups were assigned for the sake of comparison. Garlic extract (50mg/kg bw/mouse) was given orally, day after day, at a fixed daytime. Seven weeks post-infection, adult schistosomes were recovered by perfusion and the livers of the mice were excised out and were processed for DNA extraction and Random Amplification of Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR). The results showed that garlic exerted no major changes in the genome of schistosomes. Nevertheless, that schistosomal infection induced genetic alterations in the DNA of mice, and garlic was able to ameliorate such alterations to a great extent.

SEM studies on Trichurus muris from Psammomys obessus from Sinai Peninsula, Egypt.

The whipworm Trichuris muris was recovered from the caecum of the wild rodent Psammomys obessus trapped from Sinai Peninsula, Egypt. The cuticular surface ultrastructure is described using SEM. T. muris is closely related to other Trichuris species but can be distinguished from them mainly by differences in the posterior end of males. Details of the surface such as the bacillary gland, cuticular inflations and several morphological details obtained by scanning electron microscopy confirmed the characteristics that differentiate the species. P. obessus (Cretzschmar, 1828) is considered a new host record and Sinai is considered a new locality for the genus. This may through light on the spread of T. muris between Asia and Africa.

Prevalence of three liver parasites in sheep and goats in Al-Madinah Al-Munawwarah, Saudi Arabia Kingdom.

The epidemiological status of cysticercosis, hydatidosis and dicrocoelosis in sheep and the production practices of sheep farmers that increase the risk of exposure of sheep to their infectious agents were studied in Al-Madinah, during 2011-2012. A total of 450 sheep and 1650 goats were inspected randomly at an abattoir survey and the prevalence of hydatidosis, cysticercosis and dicrocoelosis was recorded as 0.4%, 13.3% & 0.6% in sheep and 0.16%, 1.8% & 0.0% in goats respectively. The prevalence of cysticercosis increased dramatically in days of wedding parties to more than 75% because a large number of sheep was brought from one grazing endemic farm. The improper disposal of dead animals, the access of farm dogs to offal of slaughtered sheep, the farmers carelessness to treat their dogs with anthelmintic, and the grazing of flocks in fields where stray dogs have free access; increased the sheep and goats risk of exposure to hydatidosis and cysticercosis.

Genetic variations among Echinococcus granulosus isolates in Egypt using RAPD-PCR.

Cystic echinococcosis (CE), caused by hydatid cysts, is a widespread and hazardous disease in humans and animals worldwide. The aim of the current study was to investigate the genetic variations among Echinococcus granulosus cyst strains isolated from sheep, camel, pig, and donkey using RAPD-PCR analysis. Seven primers of arbitrary sequences were used in the PCR reactions. The screened primers gave total patterns ranging from 27 to 39 reproducible bands for each isolate. Each population isolate gave its specific pattern. Although distinct polymorphic patterns were obtained among the four isolates, there were several shared bands among them in each primer used. A comparison of the different RAPD-PCR patterns showed that primers P1, P3, and OPH 04 yielded band patterns that revealed a high degree of divergence among the four isolates of E. granulosus that allowed easy distinction between them. The remaining primers (P2, P4, P5, and OPH14) amplified DNA fragments that were common to two or more isolates but diversified in the other two or three isolates. The study revealed that the most closely related isolates were of donkey and camel where the similarity coefficent between them ranging from 53 % to 78 %, followed by isolates of pig and sheep (sc = 40 % to 68 %), while the similarity coefficent between isolates of camel and sheep was 33-45 %, between camel and pig was 36 to 57 %, between donkey and pig was 37 to 52 %, and between donkey and sheep was 35 to 54 % which means that they more or distant from each other. In conclusion, hydatid cysts isolated from camel may have the genotypic characters of donkey strain.

Host-parasite relationships between Schistosoma mansoni and Echinostoma liei and their intermediate host Biomphalaria alexandrina using RAPD-PCR analysis.

Biomphalaria alexandrina are known to be intermediate hosts for both Schistosoma mansoni and Echinostoma liei. RAPD-PCR assay offers a new approach to host-parasite relationships. This was performed by investigating the genetic variation and compatibility among S. mansoni, E. liei and their intermediate host B. alexandrina with special emphasis on variations occurring in snails infected with S. mansoni and/or E. liei. Six primers were screened for DNA analysis and gave total patterns from 28 to 37 reproducible bands for each species. All specimens analyzed by the RAPD-PCR gave interpretable electrophoretic banding patterns that were polymorphic and compatible in the amplified products of these primers within each species.

Suface topography of adult Anchitrema sanguineum (Sansio, 1894) Looss, 1899 and Prosthodendrium (Prosthodendrium) urna (Looss, 1907) Dollfus, 1931 infecting bats in Upper Egypt (Qena).

Anchitrema sanguineum and Prosthodendrium (Prosthodendrium) urna are two digenean trematodes infecting many species of bats in Egypt. The surface topography of them was studied by scanning electron microscopy. Examination of A. sanguineum revealed the presence of posteriorly directed broadbase spines allover the body. The oral sucker is bordered by several small sucker-like papillae and few irregularly distributed pores. The ventral sucker is smaller than the oral one and surrounded by several papillae. In P. (P.) urna the body is covered with simple, spines posteriorly directed. The oral sucker has few papillae and its tegumental rim slightly elevated. The ventral sucker is slightly larger than the oral one.

Diagnosis of Fasciola gigantica in snail using the polymerase chain reaction (PCR) assay.

The 124-bp repetitive and highly abundant DNA sequence--used as a specific probe for the detection of Fasciola hepatica infection in snails--was tested in the detection of F. gigantica infection in Lymnaea natalensis. The probe did not show any positive PCR results with Lymnaea natalensis, Physa acuta, Biomphalaria alexandrina and Bulinus trucatus or with Schistosoma mansoni, S. haematobium and Echinostoma liei. However, the probe was found capable to detect F. gigantica infection within L. natalensis at very early stages of the prepatent period and at very low concentrations. Thus, the present assay is specific and sensitive for the detection of F. gigantica within its intermediate host. It confirmed the idea that 124-bp repetitive and highly abundant DNA sequence in Fasciola sp. genome could be used as an epidmiological tool for examination of fasciolosis intermediate host. The nucleic acid-based assay could eliminate both inherent uncertainties and lengthy periods of time required for visual examination of the snails. Also, the assay is valuable in epizootiology of F. gigantica, vector suitability and host-parasite relationship.