Molecular identification and morphological variations of Amblyomma lepidum imported to Egypt, with notes about its potential distribution under climate change.

The tick Amblyomma lepidum is an ectoparasite of veterinary importance due to its role in transmitting livestock diseases in Africa, including heartwater. This study was conducted in 2023 to monitor Amblyomma spp. infestation in dromedary camels imported from Somalia, Ethiopia, and Sudan to Egypt. This study inspected 200 camels at the Giza governorate's camel market that had been imported from Somalia, 200 from Ethiopia, and 200 from Sudan for tick infestation. Specimens were identified using morphological characteristics and phylogenetic analyses of the 12S and 16S rRNA genes. Clusters were calculated using an unweighted pair-group method with arithmetic averages (UPGMA) dendrogram to group the specimens according to their morphometric characteristics. The morphometric analysis compared the body shape of ticks collected from different countries by analyzing dorsal features. Principal component analysis (PCA) and canonical variate analysis (CVA) were performed to obtain body shape variation among specimens from different countries. Results indicated that camels were infested by 57 males Amblyomma lepidum, and no female specimens were observed; among these specimens, one may have a morphological abnormality. The results suggest that A. lepidum specimens collected from camels imported to Egypt from African countries exhibit locally adapted morphology with variations among specimens, particularly variations in body size. This adaptation suggests minimal potential for genetic divergence. Ecological niche modeling was used to predict the areas in Africa with suitable climates for A. lepidum. The study confirmed that East African countries might have the most favorable climatic conditions for A. lepidum to thrive. Interestingly, the amount of rain during the wettest quarter (Bio16) had the strongest influence on the tick's potential distribution, with suitability decreasing sharply as rainfall increased. Future predictions indicate that the climatic habitat suitability for A. lepidum will decrease under changing climate conditions. However, historical, current, and future predictions indicate no suitable climatic habitats for A. lepidum in Egypt. These findings demand continuous surveillance of A. lepidum in camel populations and the development of targeted strategies to manage tick infestations and prevent the spread of heartwater disease.

New occurrence of Cirolana capricornica (Isopoda: Cirolanidae) from Epinepheluschlorostigma in Suez Governorate, Egypt.

Background and Aim: The isopods of the Crustacea are noteworthy. All marine, fresh, and brackish waterways at all depths are home to aquatic organisms. This order also includes species that live on land. This study aimed to report a new occurrence of the isopod Cirolana capricornica on the operculum, mouth, and body cavities of Epinephilus chlorostigma in the Suez Governorate, Egypt. Materials and Methods: With the help of fishermen, 50 live E. chlorostigma (Linnaeus, 1758) were randomly gathered along the Red Sea coast of the Suez Governorate during November and December 2019 for the current investigation. Isopods were isolated from the fish samples and captured using light and electron microscopy for morphological identification. Results: Some fish were emaciated, and minute white isopods were attached externally to the skin near the gills and mouth cavity, and internally to the mouth cavity. No correlation was observed between body cavity attachment and gross lesions. The prevalence of infestation was 16%. Conclusion: C. capricornica was identified using optical and electron microscopy to analyze the isopod specimens' morphology. This scavenging isopod species is newly discovered in Egypt.

Molecular screening of Amblyomma species (Acari: Ixodidae) imported from African countries to Egypt, with the first report of Amblyomma latum from the ball python, Python regius (Squamata: Pythonidae).

Amblyomma species are non-endemic ticks in Egypt, which have been recorded from imported animals. This study was carried out in 2022 to monitor Amblyomma spp. from dromedary camels, cattle, and snakes in Egypt. During this study, 400 camels, 200 cattle, and two snakes (Pythonidae) were inspected for tick infestation. Collected specimens were identified based on morphological characters and confirmed by phylogenetic analysis of the 12S rRNA gene. Camels were infested by adult specimens of Amblyomma variegatum and Amblyomma lepidum, but no Amblyomma spp. were collected from cattle. Amblyomma variegatum showed high genetic similarity to other A. variegatum from Guinea-Bissau and São Tomé (> 99.99%), and A. lepidum showed high genetic similarity to other A. lepidum from Israel and Sudan (99.99%). Amblyomma latum is recorded in Egypt from the ball python snake for the first time and showed high genetic similarity with South African A. latum (99.87%).

In-vivo and in-vitro effectiveness of three insecticides types for eradication of the tick Rhipicephalus sanguineus in dogs.

Background: External parasites contribute to extensive harmful impacts on their hosts which is why control and eradication of external parasites have been included in all biosecurity plans of dog houses. Aim: To evaluate the in-vitro and in-vivo effectiveness of chemicals like Doramectin injectable and Fipronil 50 mg/ml drops and herbal mixes eco-friendly insecticides like phenylpyrazole-garlic-camphor mix spray for combating the external parasitism in dogs and their influence on the hematological, biochemical, and cortisol (CORT) profiles. Method: The in-vitro effectiveness of the insecticides was conducted by using a total of 216 developmental stage Rhipicephalus sanguineus (72 adults, 72 larvae, and 72 eggs) designed into three replicates of petri dishes (3 plates × 8 units × 3 stages/replicate); each replicate was exposed to 1 ml insecticide. The number of surviving ticks was recorded after 0, 2, 4, 8, 16, and 24 hours. Sixteen Rottweiler male dogs aged 1 year and 45.5 kg were divided into four groups. Three groups (G1, G2, and G3) were experimentally infested with R. sanguineus ticks 3-4 weeks post-dog arrival and kept under observation from zero-time of experimental infestation for 1-2 weeks. The three experimentally infested dog groups were treated with Doramectin injectable, Fipronil 50 mg/ml drops, and phenylpyrazole-garlic-camphor mix spray, respectively, and the fourth group was designed as a negative control. A total of 144 samples, including 48 ethylenediaminetetraacetic acid blood, 48 whole blood, and 48 sera samples, were collected. Results: The in-vitro efficacy revealed highly significant (p < 0.01) 100% killing efficacy that was achieved after 8 hours in Doramectin and Fipronil 50 mg/ml and 24 hours in phenylpyrazole-garlic-camphor mix. The in-vivo trials revealed highly significant (p < 0.01) improvements of red blood cells, hematocrit, mean corpuscular hemoglobin concentrations, platelets, total and differential leukocytic counts, erythrocyte sedimentation rates in the second hour, total protein, creatinine, alanine aminotransferase, urea, glucose, triglycerides, total cholesterol, and CORT levels in the 2-week (P1) and 4-week posttreatment (P2) samples in Dormectin, Fipronil 50 mg/ml, and phenylpyrazole-garlic-camphor mix-treated dogs with more pronounced recovery in phenylpyrazole-garlic-camphor mix spray-treated dogs. Conclusion: The insecticides were able to provide a high level of protection against experimental infestation with concern to the different modes of application. Phenylpyrazole-garlic-camphor mix spray (eco-friendly) achieved higher insecticidal action compared to the chemicals.

Morphological and molecular studies on tick species in Ismailia Governorate in Egypt and Al Gabal Al Akhdar in Libya.

Background: Ticks are obligate blood-sucking ectoparasites of vertebrates that have an impact on both domestic and wild animals, especially in tropical and subtropical areas. Aim: The objective of this study is to compare the prevalence and the tick species in both Al Gabal Al Akhder regions in northeastern Libya and Ismailia, Egypt. Methods: Tick specimens collected from predilection sites on the hosts were identified by morphological (light microscopy) and molecular methods. Results: In Ismailia, Egypt, 23.9% of the 230 (examined cattle and buffaloes) were infested with one species of hard ticks, Rhipicephalus annulatus. In the Libyan province of Al Gabal Al Akhdar, the prevalence of tick infestation in cattle, sheep, and goats, was 47% and 59%, respectively. R. annulatus is the identified tick species for cattle, Hyalomma marginatum, and Rhipicephalus bursa, are the identified tick species of the infested sheep and goats. Conclusion: The 16S rDNA gene sequencing and phylogenetic analysis of sample species from Egypt and Libya proved instrumental in overcoming the difficulties associated with morphological identification techniques.

Comparison of Some Molecular Markers for Tick Species Identification.

BACKGROUND: Ticks are obligate blood-sucking ectoparasites of vertebrates. Since many tick identification studies are based on the analysis of 16S rDNA, 12S rDNA and ITS-1, 2 rDNA genes, we aimed to compare the performance of these molecular markers of common use for the identification of ticks, under a diagnostic laboratory environment. METHODS: Overall, 192 tick specimens were collected through the state of Texas from January 2014 to August 2015 and the species was determined by both morphology and molecular amplification using the 16S rDNA, 12S rDNA, ITS1 and ITS2. RESULTS: The species collected were identified by molecular techniques as Dermacentor albipictus, D. variabilis, Amblyomma americanum, Ixodes scapularis, A. cajennense, Rhipicephalus sanguineus and Carios capensis. ITS1 and ITS2 were not able to prove consistent amplification and therefore have been considered as potential markers for tick identification. CONCLUSION: The use of mitochondrial genes in tick identification showed to provide more consistent results in the diagnostic environment.

Effectiveness of poultry litter amendments on bacterial survival and Eimeria oocyst sporulation.

AIM: Broilers' optimum performance in response to their genetic potential depends on litter environment which is ideal for bacterial survival and coccidian oocyst sporulation. An in vitro evaluation was conducted for the effectiveness of superphosphate, meta-bisulfide, and charcoal litter amendments in minimizing Escherichia coli O157:H7 and Salmonella Typhimurium survival, Eimeria oocyst count, and sporulation. MATERIALS AND METHODS: Three groups of 16 litter trays were prepared and inoculated with E. coli O157:H7, S. Typhimurium, and Eimeria non-sporulated oocyst. A set of four trays in each group was designed for each one of the chemical amendments. A total of 720 litter samples were collected and examined for bacterial counts, Eimeria oocyst count, and sporulation during the experimental period (35 days). RESULTS: Litter moisture and pH revealed a highly significant (p<0.001) reduction in all treated litter trays compared to control. Total bacterial count (TBC), total Enterobacteriaceae count, and S. Typhimurium count showed a highly significant (p<0.001) reduction in meta-bisulfide-treated trays compared to other amendments and positive control. Meanwhile, Eimeria oocyst count and sporulation revealed a highly significant (p<0.001) reduction in superphosphate, meta-bisulfide, and charcoal-treated trays, respectively. Temperature revealed a highly significant (p<0.001) weak positive correlation with pH of all inoculated trays, a highly significant (p<0.001) weak negative correlation with moisture percentage of E. coli O157:H7 and S. Typhimurium inoculated trays, and a highly significant (p<0.001) weak negative correlation with TBC. Meanwhile, relative humidity revealed significant (p≤0.005) weak positive correlation with moisture percentage of E. coli O157:H7 inoculated trays. CONCLUSION: The study concluded that regular usage with periodical reapplication of litter amendments as meta-bisulfide or superphosphate in poultry farms is one of the indispensable managemental and preventive measures for minimizing bacterial survival and inhibiting Eimeria oocyst maturation and sporulation.

Prevalence of Eimeria species among sheep and goats in Suez Governorate, Egypt.

Coccidiosis is a disease of high economic importance caused by Eimeria species that show ubiquitous distribution among several species including small ruminants. The prevalence of Eimeria infection in sheep and goats in Geneffe village, Suez Governorate, Egypt was determined during the period from March 2015 to February 2016. Total of 277 animals (142 sheep and 135 goats) were clinically examined and fecal samples were collected and tested both microscopically and by PCR. Sera samples of sheep and goats under 1 year were collected for biochemical analysis. Results revealed that (60%) of goats and (57.70%) of sheep were suffering from subclinical coccidiosis. Adult female goats were significantly (P < 0.05) more infected (82.2%) than adult male goats (40%). Eimeria infection was significantly prevalent in summer (75%) and autumn (74.2%) in sheep than winter (38.2%) and spring (43.2%), while goats did not show significant seasonal variations of infection. The Eimeria species were identified as E. crandallis, E. granulosa, E. ovina, E. parva, E. faurei, E. ovinoidalis, E intricate, E. pallida, E. arloingi, and E. ahasta in sheep, and E. ninakohlyakimovae, E. hirci, E. caprina, E. christenseni, E. jolchijevi, E. apsheronica and E. arloingi in goats. Although animals were subclinically infected with coccidia, some significant biochemical changes were observed in serum samples of sheep and goats. The molecular detection of Eimeria oocysts did not yield any positive results but after sporulation, Eimeria oocysts were detected at zone 100 bp. Our results showed a moderate prevalence of Eimeria infection among adult and yearling sheep and goats in Geneffe village. Suez governorate, Egypt.Hence, good control and prevention programs are necessary.