Genetic diversity and population structure of Echinococcus multilocularis: An in-silico global analysis.

Objective: Alveolar echinococcosis is caused by Echinococcus multilocularis, a parasite of zoonotic significance with a wide range of intermediate and final hosts, and the parasite survives successfully in diversified conditions. Plentiful studies have been done to study the genetic structure of the population of the parasite and the level of intimate kinship using mitochondrial (mt) DNA. The present study was conducted to investigate the population structure, genetic variation, and phylogenetic relationship of various isolates of E. multiocularis submitted to GenBank worldwide. Sequences of mt genes (mt-cytochrome c oxidase (cox1), mt-NADH dehydrogenase (nad1)) of E. multilocularis were analyzed to achieve the set goals. Materials and Methods: A total of 275 and 124 gene sequences of mt-cox1 and mt-nad1 belonging to E. multilocularis, respectively, were retrieved from the National Center for Biotechnology Information GenBank. The retrieved sequences were subjected to alignment with respective reference sequences using MEGA software. The PopArt software was used to establish median-joining networks, while DnaSp was used to calculate neutrality and diversity indices. MrBayes software was used to investigate the phylogenetic association between haplotypes based on Bayesian phylogeny. Results: Approximately 13 and 20 distinctive haplotypes of nad1 and cox1 genes, respectively, were observed in the present study. In both of the mt genes, diversity indices indicated low haplotype (mt-cox1 = 0.140; mt-nad1 = 0.374) and nucleotide (mt-cox1 = 0.00111; mt-nad1 = 0.00287) diversities. The values of Tajima's D and Fu Fs for a population of both of the genes under study were found to be negative. Conclusion: This study is a maiden attempt to provide insights into the population structure and genetic variation of E. multilocularis on a global scale. However, it is suggested that to better understand the population structure and genetic diversity of E. multilocularis, more geographical locations and amplifications of full-length gene sequences should be considered, which could be helpful in widening the insights into the genetic diversity of E. multilocularis.

Pathophysiological and pro-inflammatory cytokine surveys on livestock normally infected with Taenia saginata cysticercosis.

Background: Cattle and buffaloes can contract cysticercosis, an infection of the muscles brought on by Taenia saginata larvae. Despite having a global spread, cysticercosis is more prevalent in impoverished nations due to impaired hygiene standards. It has been discovered that Taenia saginata cysticercosis routine visual diagnosis is not very effective, especially in mild infections. Therefore, a more trustworthy in vivo test might be used as an alternative in slaughterhouses and epidemiological studies. Biochemical assays are possibly utilized as an alternative to detect cysticercosis inside a topical environment. Aim: Investigating serum biochemical alterations in cattle with cysticercosis was the goal of the current research. As a further method of diagnosis, it was also determined how Cysticercus bovis affected pro-inflammatory cytokines and histopathology. Methods: Blood samples from 42 slaughtered cattle (21 healthy and 21 sick animals) were taken from Assiut abattoir. Using an ELISA and spectrophotometer, respectively, their serum's pro-inflammatory cytokines and biochemical profile were evaluated. These cattle were chosen between March 2023 and February 2024. Results: A percentage of 4.6% of the 455 cattle examined after being slaughtered had T. saginata cysticerci infections. All values in the serum biochemistry were considerably different (p < 0.01), whereas the majority of biochemical parameters increased significantly (p < 0.01) in infected animals. In contrast, there was a substantial (p < 0.01) decline in HDL-c, SOD, CAT, and GSH. On the other hand, procytokine inflammatory indices for both TNF-α and IL-1ß indicated a substantial increase (p < 0.01) in infected cattle. Additionally, the histological results revealed significant alterations in the tissues of infected livestock. Conclusion: This has been inferred cysticercosis possesses negative impacts on cattle's plasma biochemical profiles, indicating the field applicability of biochemical measures in outbreaks of bovine cysticercosis. Pro-inflammatory cytokine indices and histological changes could be included as further indicators of T. saginata cysticercosis in cattle.

Global scenario of genetic diversity in cox1 and nad1 genes of Moniezia expansa.

Monieziasis is a parasite-borne production-limiting disease of livestock. Moniezia expansa is the most important species having cosmopolitan distribution. Despite of numerous prevalence reports, very little information is available about the evolutionary biology and population genetics of M. expansa. To close this research gap, this study was undertaken to recognize and inspect the genetic variation of M. expansa populations around the world using the cox1 and nad1 genes and deduce phylogenetic relationships with M. expansa populations. The cox1 and nad1 gene sequences were downloaded from the NCBI GenBank database. Followed by sequence alignment, median-joining networks were constructed using PopArt software. Diversity and neutrality indices were computed through DnaSp software while MEGA software was used to draw the maximum-likelihood phylogenetic tree. Thirty-two cox1 sequences, from five different countries, and 9 nad1 sequences from three different countries, were among the sequences used in this study. The cox1 and nad1 gene sequences had mutations in 97 and 36 different places, respectively. Twenty and 7 unique haplotypes were discovered for the cox1 and nad1 gene sequences, respectively. Comparable haplotype diversities were observed for both the genes under study (cox1 = 0.950; nad1 = 0.944). Negative Tajima's D and Fu Fs were found for the cox1 gene while these indices were positive for the nad1 gene. Phylogenetic analysis also showed the existence of unique haplotypes for both the cox1 and nad1 genes. The results of this study indicate that there is the existence of a huge genetic diversity in M. expansa isolates. For future studies, it is recommended that longer gene sequences should be used to describe genetic variation among M. expansa isolates as the length of the gene under study affects the genetic variation. Moreover, additional mitochondrial markers should also be investigated because the assertive strength of a group of gene targets is superior to defining genetic diversity.

Serosurvey and associated risk factors for Neospora caninum infection in Egyptian water buffaloes (Bubalus bubalis).

Neosporosis is a parasitic disease that causes reproductive disorders in animals, making it a barrier to maximum efficiency. The purpose of this study was to determine the seroprevalence of Neospora caninum (N. caninum) antibodies in water buffaloes from four governorates in northern Egypt. A commercial indirect-ELISA test was used to detect antibodies against N. caninum in the serum of 450 water buffaloes. The total seroprevalence of N. caninum in water buffaloes from Egypt was 31.3%, and the highest prevalence was observed in Gharbia governorate. The identified risk factors for N. caninum infections in water buffaloes were sex (OR = 1.96, 95%CI: 1.22-4.17), buffaloes more than 4 years of age ( OR = 5.80, 95%CI: 2.26-14.86), abortion in second trimester (OR = 16.48, 95%%CI: 2.99-34.03), history of abortion (OR = 3.45, 95%CI: 1.58-7.52) and contact with dogs (OR = 2.55, 95%CI: 1.51-4.32). Thus, more studies are needed to determine the role of buffaloes in the epidemiology of neosporosis in Egypt.

Seroprevalence and risk factors for Neospora spp. infection in equine in Egypt.

Neospora infections in equine are associated with reproductive disorders and neurological diseases. Nevertheless, Egypt has no epidemiological information on this parasite in equine. This study determined the prevalence of Neospora spp. infection in 325 equines from three Egyptian governorates located at Northern Egypt using cELISA. The prevalence of antibodies against Neospora spp. was 19% (95% CI: 14.09-25.05) in horse, 34.1% (95%CI: 24.92-44.69) in donkey and 26.7% (95% CI: 15.97-41.04) in mule. In comparison to horse, donkey had a considerably higher chance of contracting Neospora spp. infection (OR = 1.80, 95% CI: 0.78-4.13; P = 0.016). The risk was also higher in freely grazing animals (OR = 3.49, 95% CI: 0.95-12.78; P = 0.059). Moreover, yearling animals (12-24 months) (OR = 5.03, 95% CI: 1.51-16.80; P = 0.009) and those with natural breeding (OR = 11.80, 95% CI: 3.24-42.99, P < 0.0001) and a history of early abortion (OR = 7.04, 95% CI: 3.01-16.47; P < 0.0001) also showed a greater risk of seropositivity. The risk of Neospora infection increased significantly in equines contacted with dogs (OR = 5.16, 95% CI: 1.76-15.10; P = 0.003). This the first serological study to determine the seroprevalence of Neospora spp. in equine in Egypt. Further studies are necessary to identify the species of Neospora and to understand the role of above-mentioned risk factors in spreading of infection.

Prevalence of Cryptosporidium infection and associated risk factors in calves in Egypt.

Cryptosporidium is one of the causative parasitic agents that causes gastrointestinal diseases in calves. The parasite poses a zoonotic risk to immunocompromised individuals and children. Thus, this study aimed to determine the prevalence of Cryptosporidium infection in calves in three Egyptian governorates situated in Nile Delta and assess the associated risk factors. The Cryptosporidium oocysts were detected in 81 out of 430 calves (18.84%). In addition, the univariant analysis showed that age, feeding source, hygienic status, presence of diarrhea and contact with other animals were significantly (P < 0.05) associated with Cryptosporidium prevalence in calves. Furthermore, the risk factors related with Cryptosporidium prevalence were age (OR 1.96, 95%CI 0.97-3.94), feeding on milk and pasture (OR 2.07, 95%CI 1.15-3.72), poor hygienic condition (OR 2.25, 95%CI 1.28-3.94), presence of diarrhea (OR 2.47, 95%CI 1.23-4.96) and contact with other domestic animals (OR 2.08, 95%CI 1.24-3.50). In addition, the PCR assay targeting 18srRNA showed that the most prevalent species among calves was C. parvum. Although additional researches are required to understand the most effective steps that farmers and veterinary professionals should take to decrease the occurrence of Cryptosporidium infection.

Neospora caninum infection in dairy cattle in Egypt: a serosurvey and associated risk factors.

Neospora caninum (N. caninum) is one of the causative agents that causing cattle abortion, and severe economic losses. Due to the scarcity of data on N. caninum infection in Egyptian cattle, the purpose of this study was to estimate the seroprevalence and determine the risk factors for parasite infection. In four governorates in northern Egypt, 540 blood samples from cattle were taken, and tested using a commercial ELISA kit. The overall seroprevalence of N. caninum in examined cattle was 28.89%. A multivariate logistic regression model determined that age (OR = 2.63, P < 0.001), manual milking (OR = 1.39, P = 0.14), abortion history (OR = 2.78, P < 0.0001), repetition of estrus (OR = 2.31, P < 0.0001), and contact with dogs (OR = 2.57, P < 0.0001) were significant risk factors. The findings proved that N. caninum infection was one of the factors contributing to abortion and financial losses in dairy cattle in Egypt. Therefore, the application of sanitary security and control programs is very important in dairy farms.

Seroprevalence and associated risk factors for Toxoplasma gondii in water buffaloes (Bubalus bubalis) in Egypt.

Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii and affecting all warm-blooded animals. The available data about the epidemiological situation of T. gondii in water buffaloes in Egypt are scarce. Thus, a cross-sectional study was conducted to determine the seroprevalence of T. gondii in water buffaloes in three Egyptian governorates and to evaluate the associated risk factors for the infection. A total of 430 sera samples were examined using commercial Indirect ELISA Multi-species kit. The overall seroprevalence rate of T. gondii in examined water buffaloes was 7.4 %, and the highest rate (9.3 %) was found in Kafr ElSheikh governorate. Multivariate logistic regression analysis showed that adult buffalo (OR = 7.10; 95 % CI: 0.87-57.68; P = 0.067) and small herds (OR = 8.42; 95 % CI: 1.07-66.02; P = 0.043) were more likely than young buffalo and large herds to become infected with T. gondii. Moreover, the risk of buffaloes contracting T. gondii infection was higher in winter and especially among animals contacted with cats. It is necessary to identify risk factors in order to determine what mitigation, control, and prevention strategies to implement in order to reduce, control, and prevent T. gondii infection in domestic animals, which will in turn reduce human infection with the disease.

Molecular prevalence and associated risk factors of Theileria annulata infections in dromedary camels in Egypt.

Theileria annulata (T. annulata) is an intracellular protozoan, transmitted by ixodid ticks of the genus Hyalomma and affects camels. There are few epidemiological data on T. annulata infection and its associated risk factors in Egyptian camels. Thus, the present study aimed to determine the prevalence of T. annulata in camels using PCR and assess the associated risk factors for infection. A total of 380 blood samples were collected from camels raising in three Egyptian governorates and examined by PCR assay targeting 30-kDa gene to detect the presence of T. annulata infection, beside statistical analysis of associated factors. The results revealed presence of T. annulata with overall prevalence of 21.1%. In addition, the univariate analysis revealed significant (P<0.05) association between prevalence of T. annulata in camels and locality, age, sex, tick infestation, and application of acaricides. Whereas the prevalence of T. annulata was higher in camels of age group >6 to 10 years (38%), females (25.7%) and in infested camels with ticks (29%) and in case of absence of acaricides application (25%). In contrast, the body condition of camels had not significant effect on prevalence of theileriosis in camels. The current study concluded that T. annulata is prevalent in Egyptian camels and that a tick control program is required to reduce the risk of infection.

Fasciola hepatica Infection in Horses in Three Governorates in Northern Egypt: Prevalence and Risk Factors.

Fasciola hepatica is a trematode parasite that affects wide range of mammalian hosts including horses. There are no epidemiological data on fasciolosis in Egyptian horses. Hence, the present study aimed to evaluate the seroprevalence of F. hepatica in horses using ELISA as well as assess the associated risk factors for fasciolosis infection. A total of 400 sera samples of horses from three Egyptian governorates were examined. The overall seroprevalence rate in horses was 13% and the disease was more prevalent in Qalubia (14.17%) and Gharbia (16.67%) which are located at Nile Delta. The likelihood of having F. hepatica seropositivity was higher in mixed-breed animals and stallions (OR = 3.97, 95%CI: 1.10-14.28 and OR = 2.07, 95%CI: 1.09-3.93, respectively), and in older horses more than 10 years (OR = 5.69, 95% CI: 1.98-16.36). Horses were more likely to be seropositive with F. hepatica when kept in outdoor housing (OR= 1.53, 95%CI: 0.78-3), during autumn (OR = 3.80, 95%CI: 1.17-12.32) and in the presence of snails (OR = 2.72, 95%CI: 1.34-5.52). Therefore, the obtained data contributes to the understanding of the epidemiology of Fasciola species, assess in establishment of management programmes to reduce disease and public health risks.

Revealing novel cytb and nad5 genes-based population diversity and benzimidazole resistance in Echinococcus granulosus of bovine origin.

Cystic echinococcosis (CE) is a neglected zoonotic disease caused by Echinococcus granulosus (sensu stricto). The parasite affects a wide range of livestock and wild animals. In this study, the population diversity of the Echinococcus species was investigated based on mitochondrial cytochrome b (cytb) and NADH dehydrogenase subunit 5 (nad5) genes. In addition to this, ß-tubulin gene isoforms of Echinococcus granulosus were amplified to determine the resistance against benzimidazoles. For this purpose, 40 cyst samples from cattle (n = 20) and buffaloes (n = 20) were collected from the main abattoir of Sialkot. DNA extraction was performed using Qiagen Blood and Tissue Kits. Amplification was performed through PCR. Each amplicon was confirmed by GelRed™ stained agarose gel (2%). Samples were sequenced in a DNA analyzer and viewed for any misread nucleotide by using MEGA (v.11). Corrections in nucleotide sequence and multiple sequence alignment were made through the same software. NCBI-BLAST was used for sample specific sequences to identify them as belonging to a particular species. Diversity indices were estimated using DnaSP (v.6) while phylogenetic analysis was inferred using the Bayesian method using MrBayes (v.1.1). ß-tubulin gene isoforms sequence analysis was performed to find out the candidate gene causing benzimidazole resistance. All 40 isolates were found positive for E. granulosus. BLAST-based searches of sequences of each isolate for each gene (nad5 and cytb) confirmed their maximum similarity with the G1 genotype. Overall, high haplotype diversity (Hd nad5 = 1.00; Hd cytb = 0.833) and low nucleotide diversity (π nad5 = 0.00560; π = cytb = 0.00763) was identified based on diversity indices. For both the genes, non-significant values of Tajima's D (nad5 = -0.81734; cytb = -0.80861) and Fu's Fs (nad5 = -1.012; cytb = 0.731) indicate recent population expansion. Bayesian phylogeny-based results of nad5 and cytb sequences confirmed their genotypic status as distinct from other Echinococcus species. This study shed light on the status of benzimidazole resistance in Echinococcus granulosus for the very first time from Pakistan. The findings of this study will significantly add in the information available on genetic diversity of Echinoccous granulosus based on cytb and nad5 genes sequences.

Antihelimentic effect of Androctonus crassicauda scorpion venom against Trichuris arvicolae isolated from Psammomys obesus in Egypt.

Trichuridae family has a genetic and morphological variability between species affecting rodents, but it is considerably hard to morphologically diagnose species within the genus of Trichuris and the individuals of these species are identified according to their host, as it is known that Trichuris spp. is strictly host-specific. However, some species lack host specificity. So, it is necessary to use molecular data in order to well identify the Trichuris spp. in Egyptian rodents. The host examined in the current research is Psammomys obesus and the molecularly identified species from its cecum is Trichuris arvicolae. In addition, Trichuris arvicolae was subjected to in vitro treatment with Androctonus crassicauda Crude Venom as a model of natural alternative treatment for gastrointestinal nematodes that increasingly develop anthelmintic drug resistance. The changes in Trichuris arvicolae were monitored using scanning electron microscopy, Androctonus crassicauda Crude Venom made a significant ultrastructural surface changes in Trichuris arvicolae, including marked cuticular sloughing, disintegrated bacillary glands, bursting of vulva and edema of anal region. This study was done for closer identification of Trichuris spp. infecting rodents in Egypt and evaluating the efficacy of Androctonus crassicauda Crude Venom in vitro.

Toxoplasma gondii infection in Egyptian domestic sheep and goats: seroprevalence and risk factors.

Toxoplasma gondii is an apicomplexan protozoan parasite that has been associated with reproductive failure in small ruminants. Although T. gondii infections in ruminants and humans have been recorded in several Egypt's governorates, but little is known about the risk factors associated with T. gondii infections. In this study, 350 sheep and 290 goat serum samples from three governorates in Egypt were examined for presence of antibodies against T. gondii, and evaluate the associate risk factors for the infection. The seroprevalence in sheep and goats was 24% and 38.28%, respectively. In sheep and goats, age, sex, the presence of cats, and hygienic conditions were identified as risk factors for T. gondii infection. In addition, the prevalence rates were significantly higher in older animals more than 2 years old, females, among animals contacted with cats, and animals living in bad hygienic condition. In conclusion, sheep and goats in the examined regions are commonly infected with T. gondii. The identification of risk variables defines the sort of actions to be implemented in order to decrease, and prevent T. gondii infection in small ruminant animals and, as a result, human infection.

Prevalence, risk factors and first record of mitochondrial cox1 gene-based molecular characterization of Paramphistomum epiclitum from Pakistan.

Parasitic infestations are one of the major threats to the livestock industry in Pakistan. These have a negative impact on the production of domesticated livestock species. Paramphistomes belong to the superfamily Paramphistomoidea and are involved in infecting ruminants all over the world. To date, there was no information on mitochondrial DNA-based molecular characterization of Paramphistomum epiclitum from Pakistan. To close this research gap, this study was designed to provide insights into the epidemiology of Paramphistomum species. Paramphistomum epiclitum isolates were recovered from the rumen of small ruminants slaughtered at an abattoir located in Faisalabad city and animal demographics were recorded. DNA was extracted and mitochondrial cox1 was amplified and sequenced. Prevalence was calculated along with a 95% confidence interval in various groups. The chi-square test was applied to determine the association between different variables under investigation. A phylogenetic tree was constructed based on the Bayesian method. Population diversity indices were calculated using DnaSP 4.5 software. A total of 43 mutations were observed among 7 haplotypes. Negative values of Fu's Fs values, and Tajima's D indicated population expansion. Deworming, season, and grazing were the variables that significantly correlate (p < 0.05) with the prevalence of P. epiclitum. The high prevalence of P. epiclitum demonstrates that more studies are indeed needed to further understand the prevalence and distribution of P. epiclitum in definitive and all potential intermediate hosts in addition to intraspecies variation and relationship with populations from other locations.

Prevalence of intestinal parasitic infections among university female students, Gaza, Palestine.

BACKGROUND: The intestinal parasites are still endemic among children, women, and men in Gaza Strip. OBJECTIVES: To the best of our knowledge, this is the first study of intestinal parasites among young female students of Islamic University of Gaza to report the existence and prevalence of intestinal parasites. METHODS: A total of 305 stool samples were collected from female students in all faculties and were examined by wet mount and formal ether sedimentation technique. RESULTS: This study showed that the overall prevalence of intestinal parasites was 20.6%. The detected intestinal parasites were as follows: Entamoeba histolytica/dispar (7.5%), Giardia lamblia (4.9%), Ascaris lumbricoides (0.3%), Entamoeba coli (2.6%), Dientamoeba fragilis (1.0%), and Blastocystis hominis (3.9%). Science students showed the highest prevalence for parasitic infections (35.3%), and married students (16.7%) had higher prevalence than single students (6.5%). CONCLUSION: It was concluded that female students also are under risk of gaining parasitic infection in spite of their education. It is recommended that university students should be subjected to regular medical examinations for parasitic infections.